Mean Tumor Weight Research Articles

Abstract 5357: Total abdominal ultra-rapid FLASH irradiation enhances the efficacy of PD-1 inhibition in preclinical models of ovarian cancer

Abstract Objective: Ovarian cancer often presents with advanced stage disease. Despite surgery and chemotherapy, recurrence remains inevitable and prognosis poor. Advances in immunotherapy including programmed cell death-1 (PD-1) inhibition show promise, however clinical trials show modest response rates as a single agent therapy in ovarian cancer patients. Radiation therapy is an attractive strategy to enhance the efficacy of immune checkpoint blockade in ovarian cancer due to its ability to increase tumor-infiltrating lymphocytes and enhance immune activation. While ovarian cancer is known to be radiosensitive, total abdominal radiation is not routinely utilized due to high abdominopelvic toxicity. We have recently shown that ultra-high dose rate FLASH irradiation reduces intestinal radiation-induced toxicity. Importantly, we demonstrated efficient tumor control in preclinical models of ovarian cancer peritoneal metastasis. Here we sought to determine the efficacy and safety of FLASH irradiation combined with PD-1 inhibition in preclinical models of ovarian cancer. Methods: Female C57BL/6 mice were intraperitoneally inoculated with ID8-Asc or UPK10 ovarian cancer cells. Total abdominal FLASH irradiation at 14 Gy was administered 10 days post-inoculation. Anti-PD-1 and isotype control antibodies were injected on days 7, 10, and 13 post-inoculation. Mice were sacrificed 22-27 days post-inoculation for tumor burden and immune cell analysis. Stool count, stool weight, complete blood count, and survival were evaluated to compare acute and chronic toxicity as well as efficacy in the treatment arms. Results: Four cohorts of mice were analyzed: Isotype control antibody (IgG), anti-PD-1 (aPD-1), FLASH + IgG, and FLASH + aPD-1. In the ID8-Asc model, irradiated mice showed a 44% decrease in solid stool production with subsequent recovery to baseline by 14 days post-treatment. There was no evidence of hematopoietic toxicity in any of the treatment arms. Survival analysis is ongoing. Exploratory necropsy demonstrated significant tumor burden control in the FLASH cohorts, and in particular, a 25-fold decrease in tumor number in the FLASH + aPD-1 cohort compared to control (mean tumor weight 1.82 mg vs. 357.2 mg, respectively). FLASH + aPD-1 increased intratumoral CD4+ and CD8+ T cells and correlated with enhanced tumor control compared to FLASH or aPD-1 single agent treatments. Similar results were observed in the UPK10 model. Conclusion: Combination FLASH and PD-1 inhibition enhanced tumor control and recruitment of tumor infiltrating lymphocytes compared to IgG, aPD-1, and FLASH treatments alone, while demonstrating safety and tolerability. Our data identify a new opportunity for FLASH and checkpoint inhibition in the treatment of ovarian cancer. Citation Format: Stephanie Chow, Joshua T. Eggold, Karen Levy, Jinghui Wang, Rakesh Manjappa, Dylan Y. Breitkreutz, Amy S. Yu, Karl Bush, Oliver Dorigo, Billy W. Loo, Erinn B. Rankin. Total abdominal ultra-rapid FLASH irradiation enhances the efficacy of PD-1 inhibition in preclinical models of ovarian cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5357.

Evaluation of anticancer activities of Poria cocos ethanol extract in breast cancer: In vivo and in vitro, identification and mechanism

Ethnopharmacological relevancePoria cocos Wolf (P. cocos), a well-known traditional East-Asian medicinal and edible fungus, is one of the most important components in Chinese medicine formulas like “Guizhi fuling wan” to treat hyperplasia of mammary glands and breast cancer. Aiming of studyIn this study, we attempted to verify the anticancer efficacy of the ethanol extract of P. cocos (PC) on the breast cancer as well as to investigate its most active compound and its underlying molecular mechanism in vivo and in vitro. Materials and methodsThe key anti-cancer components were separated and purified through chromatography and identified by spectral analyses. The in vivo anti-breast cancer efficacy and side effects of PC were evaluated in BALB/c nude mice that have been subcutaneously injected with breast cancer cells MDA-MB-231. Cytotoxicity, apoptosis and cell cycle arrest of PC were evaluated in vitro by cell viability assays and flow cytometry. The protein levels were examined via western blotting. ResultsPachymic acid (PA), separated and identified as the most active compound, induced the significant cytotoxicity on breast cancer cells MDA-MB-231(IC50 value, 2.13 ± 0.24 μg/mL) and was not active against the normal breast epithelium cells MCF-10A. The in vivo experiment revealed that PC could significantly inhibit the tumor development and the final mean tumor weight of the mice in the PC group (0.51 ± 0.12g) was significantly lower than that in the model group (1.22 ± 0.45g). Notably, compared to the first-line anticancer drug cisplatin, PC showed less side effects on the function of the vital organs and the muscle strength of the mice. Among in vitro study, PC significantly inhibited the cell growth of MDA-MB-231 by inducing cell apoptosis and cell cycle arrested at G0/G1 phase in a dose-dependent manner. The expression of cell cycle-associated cyclin D1, cyclin E, CDK2, and CDK4 were downregulated, while p53 and p21 expression were upregulated following the PA treatment. In addition, PA downregulated the apoptotic regulator Bcl-2, increased the expression of pro-apoptotic protein Bax, and promoted the release of cytochrome c and the activation of cleaved caspase-3, -9 and caspase -8 via mitochondria-mediated and death receptor-mediated signaling pathways. ConclusionThis study verified the anticancer efficacy of PC on breast cancer in vivo and in vitro through induction of cell apoptosis and G0/G1 cell cycle arrest. The data also suggested that PA could be developed as an efficacious agent for breast cancer treatment with less side effects.

Effects of fucoidan on gut flora and tumor prevention in 1,2-dimethylhydrazine-induced colorectal carcinogenesis

Colorectal cancer (CRC) is one of the major malignancies in humans. This study was designed to evaluate the effects of fucoidan on gut flora and tumor prevention in 1,2-dimethylhydrazine-induced colorectal carcinogenesis in rats. We found that dietary fucoidan treatment decreased the tumor incidence and mean tumor weight and increased cell apoptosis. Fucoidan treatment decreased the expression of β-catenin C-Myc, CyclinD1 and Survivin, while the Hippo pathway was activated with increased phosphorylation levels of mammalian sterile 20-like kinase 1 and 2, large tumor suppressor 1 and 2, and Yes-associated protein. Compared with the model group, the levels of interleukin (IL)-17 and IL-23 were decreased, but the levels of interferon-γ, IL-4 and IL-10 were increased, in the fucoidan group. Fucoidan treatment increased natural killer cells in peripheral blood and the proportion of CD4+ T cells. Immunofluorescence detection of colorectal tumor tissues showed decreased expression of Foxp3 and up-regulated expression of CD68 in the fucoidan group. Moreover, fucoidan treatment decreased the levels of diamine oxidase and lipopolysaccharides and up-regulated the levels of tight junction proteins. 16S rDNA high-throughput sequencing revealed that fucoidan treatment decreased the abundance of Prevotella and increased the abundance of Alloprevotella. Fucoidan increased the levels of butyric acid and valeric acid compared to the model group. This study provides experimental evidence that dietary fucoidan may prevent colorectal tumorigenesis by regulating gut microecology and body immunity. Meanwhile, fucoidan activated the Hippo pathway and down-regulated the β-catenin pathway to induce tumor cell apoptosis and suppress tumor growth.

PD47-06 INTRATUMORAL ADMINISTRATION OF A RECOMBINANT BCG OVEREXPRESSING A STING AGONIST CAUSES INCREASED EXPANSION OF IFN-γ PRODUCING CD4 + T CELLS IN SOLID TUMORS

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology IV (PD47)1 Apr 2020PD47-06 INTRATUMORAL ADMINISTRATION OF A RECOMBINANT BCG OVEREXPRESSING A STING AGONIST CAUSES INCREASED EXPANSION OF IFN-γ PRODUCING CD4+ T CELLS IN SOLID TUMORS Alok Singh*, Monali Praharaj, Kara Lombardo, Max Kates, David McConkey, William Bishai, and Trinity J. Bivalacqu Alok Singh*Alok Singh* More articles by this author , Monali PraharajMonali Praharaj More articles by this author , Kara LombardoKara Lombardo More articles by this author , Max KatesMax Kates More articles by this author , David McConkeyDavid McConkey More articles by this author , William BishaiWilliam Bishai More articles by this author , and Trinity J. BivalacquTrinity J. Bivalacqu More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000934.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical BCG causes an increase in the the CD4+ T cell population. We previously demonstrated a lack of T cell phenotype differentiation using gene expression profiling of NMIBC after BCG instillation. Intratumoral administration of STING agonists results in tumor regression in part due to tumor-specific adaptive immune responses, but BCG strains modified to overexpress STING agonists remain largely unexplored in NMIBC. We hypothesized that intravesical rBCG overexpressing a STING agonist may potentiate antitumor T cell immunity. We generated a STING agonist (c-di-AMP) overexpressing BCG (BCG-disA-OE) to test this hypothesis using an immunocompetent MB49 subcutaneous implantation model in mice. METHODS: Subcutaneous MB49 urothelial tumors were generated by implanting 0.3 x106 MB49 cells (n=9) in C57BL/6 mice. 3 doses of 1 x 107 BCG bacilli (WT-TICE BCG and BCG-disA-OE) were given intratumorally. Infiltrating T cell responses were assessed using flow cytometry. Systemic immune response was determined by splenic T cell responses. Immune correlates were established by measuring tumor weight regression. RESULTS: Increased regression of mean tumor weight was observed in tumors treated with BCG-disA-OE (0.06 g) as compared to WT-BCG (0.124 g). Enhanced antitumor efficacy of BCG-disA-OE over WT-BCG was accompanied by increased infiltration of IFN-γ producing CD4+ T cells in tumors (18.79% in WT-BCG vs. 32.28% BCG-disA-OE). A modest increase in IFN-γ producing splenic CD4+ T cells (7.8% in BCG-disA-OE vs 6.4% in WT-BCG) but a robust increase in splenic CD8+ T cells in BCG-disA-OE treated animals (32.2% in rBCG-disA-OE vs 26.6% in WT-BCG) suggests involvement of both T cell types in mounting a global IFN-γ response following BCG exposure. Notably, no differential change in animal body weights was observed following treatment with both BCG strains. CONCLUSIONS: Superior antitumor efficacy of BCG-disA-OE is conferred by potentiating an expansion of IFN-γ producing CD4+ T cells. Our results suggest that activation of STING signaling by recombinant BCG overexpressing STING is a promising approach to enhance the antitumor immune responses and may be valuable as an adjuvant treatment of patients with high risk NMIBC. Source of Funding: Maryland Tedco, Willowcroft Foundation, and Cigarette Restitution Fund © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e927-e927 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alok Singh* More articles by this author Monali Praharaj More articles by this author Kara Lombardo More articles by this author Max Kates More articles by this author David McConkey More articles by this author William Bishai More articles by this author Trinity J. Bivalacqu More articles by this author Expand All Advertisement PDF downloadLoading ...

The MODULATORY EFFECTS OF BONE MARROW-DERIVED MESENCHMAL STEM CELLS ON CYCLOPHOSPHAMIDE INDUCED HEPATOTOXICITY IN CARCINOMA MICE

Objective: Cyclophosphamide (CPA) is a chemotherapeutic agent, induces hepatotoxicity as one of its side effects. Therefore, the present work was designed to investigate the protective role of bone marrow-derived mesenchymal stem cells (BM-MSCs) on CPA–induced hepatotoxicity in Ehrlich Ascites Carcinoma bearing-mice (EAC) and to test whether BM-MSCs influences the antitumor properties of the CPA.
 Methods: The hepatoprotective effects of BM-MSCs (single dose of 100 µl of a cell suspension containing allogenic BM-MSCs, i. v.) was evaluated in a model of hepatotoxicity by CPA (10 mg/kg/d i. p.) in EAC-female mice for one month. The anti-tumor activities of CPA and BM-MSCs were assessed by measuring mean tumor weight, mean survival time and the increase in life span. Moreover, ALT, AST, GGT, MDA, GSH, SOD, IL-6, IL10, caspase-3 and Bcl2 were measured.
 Results: The i. p. administration of CPA and BM-MSCs resulted in significant reductions in tumor size and mean tumor weight as well as caused concurrent significant increases in the life span as compared to the EAC mice. Furthermore, BM-MSCs ameliorated the liver enzyme markers namely ALT, AST, GGT, and hepatic oxidative stress through inhibition of MDA level that correlated with significant improvement in antioxidant status via increasing GSH and SOD levels as compared to both EAC and EAC+CPA groups. Moreover, BM-MSCs treatment significantly reduced the inflammatory marker level IL-6 as well as increment the level of IL-10 with subsequent decreases apoptosis via a depletion in the caspase-3 associated with an enhancement in the level of Bcl2 as compared to EAC group and EAC+CPA group. Minor histological lesions were observed in the liver tissue sections of mice treated with CPA and BM-MSCs as compared to the high histological lesions observed in the liver of the EAC group and CPA treated group.
 Conclusion: These results concluded that the combination treatment of BM-MSCs with CPA exhibited promising potential antitumor efficacy with greater safety than CPA treatment alone in mice via its antioxidant, anti-inflammatory and antiapoptotic effects.

MiR-509-3 augments the synthetic lethality of PARPi by regulating HR repair in PDX model of HGSOC

BackgroundPARP inhibitors have been the most promising target drugs with widely proven benefits among ovarian cancer patients. Although platinum-response, HR-related genes, or HRD genomic scar detection are acceptably used in assessment of Olaparib response, there are still evident limitations in the present approaches. Therefore, we aim to investigate more accurate approaches to predict Olaparib sensitivity and effective synergistic treatment strategies.MethodsWe probed two databases (TCGA and Qilu Hospital) in order to quest novel miRNAs associated with platinum-sensitivity or HR-related genes. Cellular experiments in vitro or in vivo and PDX models were utilized to validate their role in tumor suppression and Olaparib sensitizing. Furthermore, HR gene mutation was analyzed through WES to explore the relation between HR gene mutation and Olaparib response.ResultsHigh miR-509-3 expression indicated better response to platinum and longer progression-free and overall survival in two independent ovarian cancer patient cohorts (high vs. low miR-509-3 expression; PFS: TCGA P < 0.05, Qilu P < 0.05; OS: TCGA P < 0.05, Qilu P < 0.01). MiR-509-3 could impair the proliferation, migration, and invasion ability but enhance the sensitivity to Olaparib of ovarian cancer cell in vitro and in vivo by directly targeting HMGA2 and RAD51. In two PDX cases (PDX1 and PDX9), miR-509-3 could significantly increase the sensitivity to Olaparib along with the decrease of RAD51 positive rate (mean tumor weight NC + Olaparib vs. miR-509 + Olaparib; PDX1 P < 0.05, PDX9 P < 0.05). Additionally, in PDX8, miR-509-3 treatment dramatically reversed the Olaparib insensitivity (P < 0.05) by downregulating RAD51 expression. RAD51 functional detection revealed that all Olaparib sensitive cases exhibited low RAD51 positive rate (lesser than 50%) in treated groups. Furthermore, among the four HR gene mutation patients, three harbored HR core gene mutation and were sensitive to Olaparib while the remaining one with non-HR core gene mutation did not respond well to Olaparib.ConclusionsMiR-509-3 can sensitize ovarian cancer cells to Olaparib by impeding HR, which makes it a potential target in PARPi synergistic treatment. HR core gene analysis and RAD51 functional detection are prospectively feasible in prediction of PARPi response.

Downregulation of microRNA-144 inhibits proliferation and promotes the apoptosis of myelodysplastic syndrome cells through the activation of the AKAP12-dependent ERK1/2 signaling pathway

BackgroundMyelodysplastic syndromes (MDS) represent a family of hematopoietic stem cell disorders characterized by ineffective hematopoiesis. While the functions of many microRNAs have been identified in MDS, microRNA-144 (miR-144) remains poorly understood. Thus, the aim of the present study was to determine the effects of miR-144 on cell proliferation and apoptosis in MDS cells and mechanism thereof. MethodsMDS-related microarrays were used for screening differentially expressed genes in MDS. The relationship between miR-144 and A-kinase anchoring protein 12 (AKAP12) was determined by a dual luciferase reporter gene assay. Subsequently, gain- and loss-function approaches were used to assess the effects of miR-144 and AKAP12 on cell proliferation, cell cycle and cell apoptosis by MTT assay and flow cytometry. Following the induction of a mouse model with MDS, the tumor tissues were extract for evaluation of apoptosis and the expression of miR-144, AKAP12, and the relevant genes associated with extracellular-regulated protein kinases 1/2 (ERK1/2) signaling pathway and apoptosis. ResultsWe observed significantly diminished expression of AKAP12 in MDS samples. miR-144 directly bound to AKAP12 3′UTR and reduced its expression in hematopoietic cells. Downregulation of miR-144 or upregulation of AKAP12 was observed to prolong cell cycle, inhibit cell proliferation, and induce apoptosis, accompanied by increased expression of AKAP12, p-ERK1/2, caspase-3, caspase-9, Bax, and p53, as well as decreased expression of Bcl-2. The transplanted tumors in mice with down-regulated miR-144 exhibited a lower mean tumor diameter and weight, and increased apoptosis index and expression of AKAP12 and ERK1/2. ConclusionTaken together, these studies demonstrate the stimulative role of miR-144 in MDS progression by regulating AKAP12-dependent ERK1/2 signaling pathway.

Assessment of Physical Performance and Early Treatment Outcomes after Implantation of Modular Prostheses of Femoral and Tibial Shaft

Tumours of bone diaphyses often require resection followed by bone reconstruction. The use of modular pro-sthe-ses permits early limb loading and rapid improvement in physical performance. The aim of this study was to evaluate the func-tioning of patients and early treatment outcomes after the implantation of modular prostheses. The analysis covered the correlation between the extent of resection, physical performance and the number of perioperative complications. 10 patients (5 women and 5 men) with diaphyseal tumours who had modular prostheses implanted were treated at the Orthopaedic Oncology Department in Brzozów between 2014 and 2018. The mean age of the patients was 51.1 years (range: 26-63 years). Functional outcomes were assessed using the MSTS and the Karnofsky scoring system. A VAS was used to evaluate pain intensity. the extent of resection was also analysed, considering bone length and tumour weight. The mean tumour weight was 374g (150-700g). The length of the implants varied from 10 to 25 cm. The mean dura-tion of hospitalisation was 16 days (14-19 days). At 3 months following the surgery, the intensity of pain had decreased from a mean of 6.8 points to 4.2 points (a decrease of 26% from the pre-operative baseline). The MSTS showed improvement of functional performance from a mean of 10.8 points (36%) to 22.9 points (76%). The Karnofsky scores demonstrated an increase in physical performance from 47 to 67 points (20 points on average). Superficial infection of the wound developed in 1 patient. 1. The treatment of diaphyseal tumours with modular prostheses produces good functional outcomes. 2. The extent of the resection and the size of the implant have an effect on the post-operative physical performance of the patients. 3. Pre-operative evaluation of the weight of the tumour may be helpful in predicting the patient's post-operative functional status. 4. Phy-sical perfor-mance is better after the resection of femoral vs tibial tumours.

Minimally invasive parathyroid carcinoma-A missing entity between parathyroid adenoma and carcinoma: Scintigraphic and histological features.

Minimally invasive parathyroid carcinoma (MIPC) is clinically and biochemically comparable with parathyroid adenoma (PA) though histopathologically differ from PA. MIPC is an intermediate of PA and parathyroid carcinoma (PC). In literature, there is no definite criterion to diagnose MIPC. Our aim was to evaluate and characterize the imaging and biochemical parameters with histological characteristics of MIPC. Ten patients with MIPC were recruited from (single centre) Indian PHPT registry (www.indianphptregistry.com) from January 2014 to July 2018. Clinical, biochemical, imaging and histological features of MIPC patients were reviewed. The mean age of MIPC patients (n=10; 3 males) was 39.9±11.3years (range: 17-50). All patients had an elevated preoperative parathyroid hormone (iPTH) level ranging from 427 to 2138pg/mL (median: 1328). MIBI scan showed intensely avid and enlarged parathyroid tumours in all patients; LIPT in 6, RIPT in 3 and ectopic mediastinal in 1 with mean size of the tumours was 2.8±1.1cm. The mean of maximum standardized uptake value (SUVmax) of MIPC in F-18 fluorocholine PET/CT was 6.7±1.1 (range 6.0-8.3). The mean tumour weight was 12±9.5g (range: 1.09-28). All MIPC patients had identified capsular invasion in 80% and vascular invasion in 50% only but there was no local invasion, lymph nodal or distant metastasis. The mean Ki-67 labelling index was 3.2±2.7 (range 1.1-10). The study concluded that MIPC patients are less aggressive (on the basis of imaging and histopathological findings) and should be differentiated from parathyroidadenomaandcarcinoma.

PLK1 contributes to autophagy by regulating MYC stabilization in osteosarcoma cells.

BackgroundPLK1, a typical PLK protein, is the main driver of cancer cell growth and proliferation. It is an inhibitor of the protein kinases that is currently being investigated in clinical studies. It is often used as a tumor marker, as high PLK1 expression correlates with poor prognosis in cancer. Overexpression of MYC is a hallmark of many human cancers. MYC modulates the transcription of thousands of genes that required to coordinate a series of cellular processes, including those essential for growth, proliferation, differentiation, self-renewal and apoptosis. To date, functions of PLK1 and MYC on tumor are mostly studied in separate researches, and studies on their mutual crosstalk are lacking.PurposeTo investigate the mechanism of PLK1 and MYC in regulating progress of osteosarcoma.MethodsProtein level was examined using Western blot. Animal experiments were performed with female FOX CHASE severe combined immunodeficient mice. Mice were randomly divided into experimental or control groups.ResultsPLK1 or MYC promoted the proliferation of osteosarcoma cells through the autophagy pathway. PLK1 contributed to MYC protein stabilization. PLK1 inhibition enhanced MYC degradation in osteosarcoma cells. PLK1 inhibition led to a marked decline in MYC protein abundance. The representative MYC target genes were deregulated by PLK1 inhibitors. BI2536 treatment caused a significant delay in xenograft tumor growth in mice injected with U-2 OS cells subcutaneously, with lower mean tumor weight compared to the control group.ConclusionPLK1 is crucial for MYC stabilization. It promotes cell proliferation by autophagy pathway in osteosarcoma cells. Data validate PLK1 as a potential therapeutic target in osteosarcoma caused by MYC-amplified.

The Role of APAL/ST8SIA6-AS1 lncRNA in PLK1 Activation and Mitotic Catastrophe of Tumor Cells.

Tumor growth can be addicted to vital oncogenes, but whether long noncoding RNAs (lncRNAs) are essential to cancer survival is largely uncharacterized. We retrieved Gene Expression Omnibus datasets to identify lncRNA overexpression in 257 cancers vs 196 normal tissues and analyzed the association of ST8SIA6-AS1 (termed Aurora A/Polo-like-kinase 1 [PLK1]-associated lncRNA, APAL) with the clinical outcomes of multiple types of cancer from public RNA sequencing and microarray datasets as well as from in-house cancer cohorts. Loss- and gain-of-function experiments were performed to explore the role of APAL in cancers in vitro and in vivo. RNA pulldown and RNA immunoprecipitation were used to investigate APAL-interacting proteins. All statistical tests were two-sided. APAL is overexpressed in multiple human cancers associated with poor clinical outcome of patients. APAL knockdown causes mitotic catastrophe and massive apoptosis in human breast, lung, and pancreatic cancer cells. Overexpressing APAL accelerates cancer cell cycle progression, promotes proliferation, and inhibits chemotherapy-induced apoptosis. Mechanism studies show that APAL links up PLK1 and Aurora A to enhance Aurora A-mediated PLK1 phosphorylation. Notably, targeting APAL inhibits the growth of breast and lung cancer xenografts in vivo (MCF-7 xenografts: mean tumor weight, control = 0.18 g [SD = 0.03] vs APAL locked nucleic acids = 0.07 g [SD = 0.02], P < .001, n = 8 mice per group; A549 xenografts: mean tumor weight control = 0.36 g [SD = 0.10] vs APAL locked nucleic acids = 0.10 g [SD = 0.04], P < .001, n = 9 mice per group) and the survival of patient-derived breast cancer organoids in three-dimensional cultures. Our data highlight the essential role of lncRNA in cancer cell survival and the potential of APAL as an attractive therapeutic target for a broad-spectrum of cancers.

Sporamin suppresses growth of xenografted colorectal carcinoma in athymic BALB/c mice by inhibiting liver β-catenin and vascular endothelial growth factor expression.

BACKGROUNDColorectal cancer (CRC) is the third most common malignancy of the digestive tract and the fifth leading cause of cancer-related mortality in China. Sporamin, a Kunitz-type trypsin inhibitor isolated from sweet potato, is a potential anti-cancer agent with activities against a number of malignant tumor cells in vitro. The liver secretes a myriad of endocrine factors that may facilitate the growth and transformation of tumors in the development of CRC.AIMTo investigate the effects of sporamin on liver morphology and biomarkers of xenografted CRC in the liver of athymic BALB/c mice.METHODSTwenty-seven male BALB/c nude mice were randomly divided into control, vehicle, and sporamin groups. Mice in the latter two groups were intraperitoneally xenografted with LoVo colorectal carcinoma cells and intragastrically infused with saline or sporamin (0.5 g/kg body weight/d), respectively, for 3 wk. Hematoxylin and eosin (HE) staining of the sections was performed to observe morphological changes in hepatic tissue and real-time fluorescent quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression of β-catenin and vascular endothelial growth factor (VEGF) in the liver.RESULTSSporamin significantly reduced the number and weight of tumor nodules formed in the abdominal cavity. Compared with the vehicle group, the mean tumor weight (± SD) in the sporamin group was significantly reduced (0.44 ± 0.10 g vs 0.26 ± 0.15 g) and the total number of tumors decreased from 93 to 55. HE staining showed that enlargement of the nucleus and synthesis of proteins within hepatocytes, as well as infiltration of inflammatory cells into the liver, were attenuated by sporamin. Immunohistochemical staining and ELISA showed that the concentrations of β-catenin and VEGF in the liver were significantly reduced by sporamin. Compared with the vehicle group, the expression of β-catenin measured in integrated optical density units per area was reduced in the sporamin group (47.29 ± 9.10 vs 26.14 ± 1.72; P = 0.003). Expression of VEGF was also reduced after sporamin intervention from 20.78 ± 2.06 in the vehicle group to 15.80 ± 1.09 in the sporamin group (P = 0.021). Compared with the vehicle group, the concentration of β-catenin decreased from 134.42 ± 22.04 pg/mL to 109.07 ± 9.65 pg/mL after sporamin intervention (P = 0.00002). qPCR indicated that compared to the vehicle group, relative mRNA expression of β-catenin and VEGF in the liver of mice in the sporamin-treated group was significantly reduced to 71% ± 1% (P = 0.000001) and 23% ± 7% (P = 0.00002), respectively, of the vehicle group levels.CONCLUSIONSporamin down-regulates the expression and secretion of β-catenin and VEGF in the liver, which subsequently inhibits the transcription of downstream genes involved in cancer progression and angiogenesis.

A novel role of Cx43-composed GJIC in PDT phototoxicity: an implication of Cx43 for the enhancement of PDT efficacy.

In spite of initially promising responses, 5-year recurrence after photodynamic therapy (PDT) sustains high level and an increase in PDT effectiveness is needed. It has been demonstrated that gap junctional intercellular communication (GJIC) formed by Connexin (Cx)43 could improve the transfer of “death signal” between cells, thereby causing the enhancement of cytotoxicity of chemotherapeutics and suicide gene therapy. Nevertheless, whether Cx43-composed GJIC has an effect on PDT phototoxicity remains unknown. This study showed that Cx43-formed GJIC could improve PDT phototoxicity to tumor cells in vitro and in vivo. Specifically, Cx43-formed GJIC under the condition of high cellular density could improve PDT phototoxicity in Cx43-transfected HeLa cells and Cx43-expressing U87 glioma cells. This effect was remarkably inhibited when Cx43 was not expressed or Cx43-formed GJ channels were prohibited. Additionally, the presence of Cx43-mediated GJIC could decrease the mean RTV and tumor weights of xenografts after Photofrin-PDT. The improved PDT efficacy by Cx43-composed GJIC was correlated with stress signaling pathways mediated by ROS, calcium and lipid peroxide. The present study demonstrates the presence of Cx43-composed GJIC improves PDT phototoxicity and suggests that therapeutic strategies designed to upregulate the expression of Cx43 or enhance Cx43-mediated GJIC function may increase the sensitivity of malignant cell to PDT, leading to the increment of PDT efficacy. Oppositely, factors that retard Cx43 expression or prohibit the function of Cx43-mediated GJIC may cause insensitivity of malignant cells to PDT, leading to PDT resistance.

Quxie Capsule Inhibits Colon Tumor Growth Partially Through Foxo1-Mediated Apoptosis and Immune Modulation

Quxie capsule (QX), a herbal remedy used in traditional Chinese medicine, is routinely used in advanced colorectal cancer treatment in Xiyuan Hospital in Beijing, China. However, the mechanism(s) underlying the effect of QX in colorectal cancer remain unclear, which hampers the optimal use of QX for the treatment of the disease. The transcription factor forkhead box O1 (Foxo1) plays important roles in regulation of cell cycle, apoptosis, and immune response in various cancers. In this study, we examined the antitumor efficacy of QX in a mouse model of colorectal cancer and further investigated the mechanism by which QX regulated Foxo1 protein-mediated pathways. QX administered via gavage daily for 2 weeks in mice carrying CT26 mouse colon tumors resulted in significantly lower mean tumor weight (0.93 ± 0.32 g) compared with that in vehicle control-treated mice (1.57 ± 0.57 g, P <.05). Foxo1 protein expression in tumors was also higher in the QX group than that in the vehicle control group. Furthermore, QX treatment upregulated apoptotic proteins such as Fas, Bim, and cleaved caspase-3 in tumor tissue compared with those in the vehicle control group. Intriguingly, the ratios of Th1/Th2 and Th17/Treg cells and levels of T-bet protein (the key regulator of Th1 and Th2 cells) were higher while the level of Foxp3 (the key regulator of Treg cells) was lower in QX-treated mice compared to vehicle control mice, revealing that Foxo1 upregulated T-bet and downregulated Foxp3 and induced a shift in immune balance. This shift could be critical in the antitumor efficacy of QX. Furthermore, knocking down Foxo1 in human colon cancer HCT116 cells partially blocked the effect of QX-elicited antiproliferative activity. Together, these results suggest that QX exerts antitumor activity in CT26 mouse colon cancer model partially mediated by Foxo1-induced apoptosis and antitumor immune response.

Treatment of seven huge retroperitoneal sarcomas: an institutional experience

Background & Aims: Retroperitoneal sarcomas (RSs) are usually found at an advanced stage, and complete resection of the tumor with wide margins including excision of other organs is difficult. Thus, the incidence of tumor recurrence is high, but the optimal chemotherapy for this tumor has not been defined. We retrospectively analyzed seven cases of huge RSs, especially focused in chemotherapy. Materials and Methods: Between 2006 and 2017, 7 huge primary RSs were operated in our hospital. Histopathologically, liposarcoma was detected in four patients, leiomyosarcoma in two, and Ewing sarcoma in one. At the initial operation, mean tumor weight of 7 patients was 2,385 grams (range: 1,000 grams - 4,500 grams), and complete tumor resection was achieved in five patients, but incomplete tumor resection occurred in two patients. Results: The overall survival was longer (not significant) for patients with complete than incomplete tumor resection. Chemotherapy was started for 5 patients after detection of recurrent tumors and having given up operative treatment through multiple operations. But the median survival time of 5 patients after the start of chemotherapy was very short (10 months; range, 4–19 months). Conclusions: Complete tumor resection at the first surgical procedure is the only way to prevent death from local recurrence. However, chemotherapy after detection of recurrent tumors is not effective now.

High-fat diets promote colon orthotopic transplantation tumor metastasis in BALB/c mice.

High-fat diets (HFDs) are a risk factor for colorectal cancer. The present study investigated whether HFDs increase colon cancer metastasis in BALB/c mice. A total of 40 BALB/c mice were divided into four groups, including the tumor, tumor-HFD, HFD and control groups. After 3 weeks, the tumor weights and metastases were observed. The serum levels of triglyceride, total cholesterol, lapin, interleukin-6 (IL-6) and tumor necrosis factor were analyzed using ELISA. The CD34, vascular endothelial growth factor (VEGF) and angiotensin 2 (ANG2) protein and mRNA levels in tumor tissues were analyzed with immunohistochemistry and reverse transcription-polymerase chain reaction. The metastasis frequency increased in the tumor-HFD group. However, there was no difference in the mean tumor weight between the tumor-HFD and tumor groups. The serum cholesterol levels were increased in the tumor-HFD and HFD groups compared with the control group. The levels of serum IL-6 and tumor necrosis factor-α were increased in the tumor-HFD group compared with other groups. The CD34 protein level, and VEGF protein and mRNA levels were increased in the tumor-HFD group compared with the tumor group. No difference was identified between the ANG2 protein and mRNA levels in of the two groups. It was concluded that HFD increased the serum level of cholesterol and cytokines, and potentially induced tumor angiogenesis, promoting transplanted orthotopic colon tumor metastasis in BALB/c mice.

Cyclin D1 silencing impairs DNA double strand break repair, sensitizes BRCA1 wildtype ovarian cancer cells to olaparib

ObjectivePoly(ADP-ribose) polymerase inhibitors (PARPi) are active in cancer cells that have impaired repair of DNA by the homologous recombination (HR) pathway. Strategies that disrupt HR may sensitize HR-proficient tumors to PARP inhibition. As a component of the core cell cycle machinery, cyclin D1 has unexpected function in DNA repair, suggesting that targeting cyclin D1 may represent a plausible strategy for expanding the utility of PARPi in ovarian cancer. MethodsBRCA1 wildtype ovarian cancer cells (A2780 and SKOV3) were treated with a combination of CCND1 siRNA and olaparib in vitro. Cell viability was assessed by MTT. The effects of the combined treatment on DNA damage repair and cell cycle progression were examined to dissect molecular mechanisms. In vivo studies were performed in an orthotopic ovarian cancer mouse model. Animals were treated with a combination of lentivirus-mediated CCND1 shRNA and olaparib or olaparib plus scrambled shRNA. Molecular downstream effects were examined by immunohistochemistry. ResultsSilencing of cyclin D1 sensitized ovarian cancer cells to olaparib through interfering with RAD51 accumulation and inducing cell cycle G0/G1 arrest. Treatment of lentivirus-mediated CCND1-shRNA in nude mice statistically significantly augmented the olaparib response (mean tumor weight ± SD, CCND1-shRNA plus olaparib vs scrambled shRNA plus olaparib: 0.172 ± 0.070 g vs 0.324 ± 0.044 g, P< 0.05). ConclusionsSilencing of cyclin D1 combined with olaparib may lead to substantial benefit for ovarian cancer management by mimicking a BRCAness phenotype, and induction of G0/G1 cell cycle arrest.

Continuing barriers to care of Wilms tumor in a low-income country.

This study evaluates the outcome of Wilms tumor (WT) following introduction of multidisciplinary team management and patient treatment stratification by tumor histology in two referral centers in southeastern Nigeria. We analyzed histologically confirmed WT cases managed from January 2008 to June 2017. There were 45 patients, peak age incidence of 2 to 5years who presented after mean symptom duration of 4.9 months (range, 1-12 months), with mean tumor weight of 1040g (range, 350-4200g). Overall, 14 (31.1%) had unfavorable histology of WT. A total of 22 (48.9%) patients received preoperative chemotherapy, 43 (95.6%) received postoperative chemotherapy based on stage of disease and histopathology, but none received adequate radiotherapy. Of these, 19 (44.2%) patients complied with chemotherapy regimen, 15 (33.3%) were lost to follow-up and 12 (26.7%) cases relapsed. With 30 cases available for evaluation and mean follow-up duration of 23months (range, 6-80 months), the overall 5-year survival is 53.3% (16 cases). Survival in children who complied with postoperative chemotherapy was 73.7%, and abandonment-sensitive survival was 35.6%. Persisting challenges were late presentation, poor compliance to treatment, and lack of radiotherapy treatment. Multidisciplinary team management and chemotherapy based on tumor histology might have resulted in slight improvement of outcome since our last report. However, to ensure survival that may approach global benchmarks, there is need for public health measures to improve time to diagnosis, and improvement of facilities and healthcare funding to ensure compliance to all phases of standard therapy.

An experimental study on antitumoral effects of KI-21-3, a synthetic fragment of antimicrobial peptide LL-37, on oral squamous cell carcinoma

PurposeThe aim of this study was to investigate the oncolytic properties of KI-21-3, a shortened fragment of LL-37, against oral squamous cell carcinoma (OSCC) in an animal model. Materials and MethodsTwelve athymic nude mice were divided into a therapy and a control group of six animals each. In both groups, SCC-4 cells were administered extraorally into the floor of the mouth in order to create an OSCC model. In the study group, KI-21-3 was applied intravenously during the 8th and 9th weeks. The subjects in the control group were injected with phosphate buffered saline solution in the same manner. During an examination period of 12 weeks, weight control was performed twice a week. Tumor growth was further controlled volumetrically via ultrasonography once a week with regular intervals. Following sacrifice, ablated tumoral tissues were immunohistochemically evaluated in order to determine the proliferation and apoptotic properties. ResultsThe mean tumor weight in the AMP group was 0.0236 ± 0.023 g, which was 30% lower than the control group with the mean value of 0.01651 ± 0.012 g. In the control group, the approximate number of the proliferating cells per visualized field was fourfold higher compared to the therapy group. Moreover, in the control group, the number of apoptotic cells per visualized field was significantly lower compared to the therapy group. ConclusionKI-21-3 showed considerable oncolytic properties on SCC-4 carcinoma cells via antiproliferative and caspase-3 apoptotic pathway. Further investigations are necessary to clarify the dose-dependent effects of this agent.

Effect of matrine on tumor growth and inflammatory factors and immune function in Wistar rat with breast cancer

To study the effect of matrine on tumor growth, inflammatory factors and immune function in Wistar rat with breast cancer. Sixty female Wistar rats were randomly divided into control group (n=10) and the modeling group of breast cancer cell tumor-bearing rat (n=50), then the rats in modeling group were randomly divided into five groups (n=10):vehicle group, matrine low dose group (50 mg/kg), medium dose group (100 mg/kg), high dose group (200 mg/kg), and lentinan group (200 mg/kg). Except the control group, each rat in the other groups was subcutaneously inoculated 0.4 ml Walker 256 breast cancer cell suspension (5×107 cells/ml) in the right axillary. Each group was treated with corresponding drug by ig administration (10 ml/kg body weight) twice a day for 14 days. After 14 days, the blood sample was collected from ventral aorta, the tumor was removed and weighed to calculate tumor inhibitory rate. The levels of interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth factor-β (TGF-β), CD3+, CD4+, CD8+, IgG, IgM, IgA in peripheral blood were determined. The mean tumor weight of matrine low-dose, medium-dose, high-dose groups and lentinan group were (4.99±0.93) g, (4.52±0.92) g, (4.22±1.18) g and (4.52±0.92) g respectively, which were significantly lower than that in model group. There was no statistical difference on the mean tumor weight among matrine groups and lentinan group (P>0.05). After the drug intervention, the tumor inhibitory rates of matrine low-dose, medium dose, high-dose groups and lentinan group were 24.6%, 31.7%, 36.3%, and 27.9% respectively, there was no statistical difference among the four groups. The levels of IL-2, IFN-γ, CD8+ in vehicle group were lower than those of control group obviously (P<0.01), however, the levels of IL-6, IL-10, TGF-β, CD3+, CD4+, IgG, IgM, IgA were higher significantly than those of control group (P<0.01). The levels of IL-2, IFN-γ, CD8+ in matrine low-dose, medium dose, high-dose groups and lentinan group were higher than those of vehicle group obviously (P<0.01, P<0.05); while the levels of IL-6, IL-10, TGF-β, CD3+, CD4+, IgG, IgM, IgA were lower than those of model group markedly (P<0.01, P<0.05). The levels of IgM and IgA in matrine low-dose and medium-dose groups were higher than those of lentinan group obviously (P<0.01, P<0.05); the levels of IL-2, IFN-γ and IgA in matrine high-dose group were higher than those of lentinan group obviously (P<0.01, P<0.05); while the levels of IFN-γ in matrine low-dose group were lower than those of lentinan group markedly (P<0.05); the levels of IL-10 and CD4+ in matrine high-dose group were lower than those of lentinan group markedly (P<0.01, P<0.05). Matrine has an obvious antitumor action which is related to its ability to enhance cellular and humoral immunity, reduce inflammatory reaction.