Proliferation Activity Of Cancer Cells Research Articles

Design, synthesis, and biological characterization of a potent STAT3 degrader for the treatment of gastric cancer.

Gastric cancer is a common malignant tumor that threatens human health, and its occurrence and development mechanism is a complex process involving multiple genes and multiple signals. Signal transducer and activator of transcription 3 (STAT3) has been elucidated as a promising target for developing anticancer drugs in gastric cancer. However, there is no FDA-approved STAT3 inhibitor yet. Herein, we report the design and synthesis of a class of STAT3 degraders based on proteolysis-targeting chimeras (PROTACs). We first synthesized an analog of the STAT3 inhibitor S3I-201 as a ligand, using the cereblon (CRBN)/cullin 4A E3 ligase ligand pomalidomide to synthesize a series of PROTACs. Among them, the SDL-1 achieves the degradation of STAT3 protein in vitro, and exhibits good anti-gastric cancer cell proliferation activity, inhibits invasion and metastasis of MKN1 cell, and induces MKN1 cell apoptosis and arrests cell cycle at the same time. Our study shows that SDL-1 is a potent STAT3 degrader and may serve as a potential anti-gastric cancer drug, providing ideas for further development of drugs for clinical use.

Development of Irinotecan Liposome Armed with Dual-Target Anti-Epidermal Growth Factor Receptor and Anti-Fibroblast Activation Protein-Specific Antibody for Pancreatic Cancer Treatment.

Pancreatic cancer is one of the most common causes of death in Taiwan. Previous studies have shown that more than 90% of pancreatic cancer cells presented epidermal growth factor receptor (EGFR) cell marker, and this marker is thought to be important as it is related to activation of cancer cell proliferation, angiogenesis, and cancer progression. Moreover, tumor-associated fibroblasts were involved in tumor proliferation and progression. In this study, we fabricated an anti-EGFR and anti-fibroblast activation protein bispecific antibody-targeted liposomal irinotecan (BS−LipoIRI), which could specifically bind to pancreatic cancer cells and tumor-associated fibroblasts. The drug encapsulation efficiency of BS−LipoIRI was 80.95%, and the drug loading was 8.41%. We proved that both pancreatic cancer cells and fibroblasts could be targeted by BS−LipoIRI, which showed better cellular uptake efficacy compared to LipoIRI. Furthermore, an in vivo mouse tumor test indicated that BS−LipoIRI could inhibit pancreatic cancer growth up to 46.2% compared to phosphate-buffered saline control, suggesting that BS−LipoIRI could be useful in clinical cancer treatment.

ROLE OF R2TP COMPLEX IN LYMPHOMA AND ITS THERAPEUTIC POTENTIAL

The R2TP complex which comprises of RUVBL1, RUVBL2, PIH1D1 and RPAP3 in humans is known to be a specialized Co-chaperone of Hsp-90 protein. This multimeric-protein complex is involved in the assembly and maturation of several multi-subunit complexes including RNA polymerase II, small nucleolar ribonucleoproteins, and complexes containing phosphatidylinositol-3-kinase-like kinases. Since their discovery as a co-chaperone of Hsp90, the R2TP complex is involved in multitude of cellular processes including, chromatin remodelling, transcription regulation, ribonucleoprotein complex biogenesis, mitotic assembly, telomerase complex assembly, and apoptosis. Lymphoma arises from the abnormal proliferation of B-cells and the R2TP complex have been reported to play an important role in the activation of p53 and RB. Therefore, the inactivation in any of the tumor suppressor pathways can drive cells to malignancy.However, there are multiple factors which may contribute towards malignancy but the folding defects in these tumor suppressor pathways could be one of the reasons. R2TP is tightly linked with oncogenesis and its inhibition can decrease the proliferation activity of cancer cells. So, the multisubunit chaperone complex as well as its components could be promising candidates for cancer chemotherapy.

Correlation between DCE-MRI quantitative perfusion histogram parameters, apparent diffusion coefficient and Ki-67 in different pathological types of lung cancer

Objective: To explore the efficacy of dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) histogram molecular imaging index, apparent diffusion coefficient (ADC) in different types of lung cancer and explore their correlation with Ki-67. Methods: A total of 33 cases of lung cancer patients confirmed by pathology in Shaoxing People's Hospital from March 2017 to March 2018 were collected, 28 males and 5 females aged 50-85 years old, including 15 cases of squamous cell carcinoma, 12 cases of adenocarcinoma, and 6 cases of small cell carcinoma. All patients performed DCE-MRI and DWI imaging within one week before surgery or puncture. ADC values, DCE-MRI quantitative perfusion parameters by histogram metrics analysis (mean value, skewness, kurtosis, uniformity, entropy, energy, quantile) of K(trans), K(ep), V(e), and V(p) were then collected. Ki-67 expression in lung cancer tissue was detected by immunohistochemical method. One-way analysis of variance and least significant difference were used to compare the differences among the parameters of the three groups which were normal distribution and equal variances, while Kruskal-Walls test and Mann-Whitney U test were used to compare the parameters that did not conform to normal distribution or variance. Pearson correlation analysis was used to compare the correlations between quantitative perfusion histogram parameters, ADC values and immunohistochemical scores of Ki-67. Results: The Ki-67 count in small cell lung cancer(458±82, P=0.011) and squamous cell carcinoma(355±277, P=0.034)were significantly higher than that in adenocarcinoma (168±164). The correlation analysis showed that there was a significant negative correlation between ADC values and Ki-67 (P=0.018, r=-0.416). And V(e) (Q5, Q10) was negatively related to Ki-67 (P=0.017, r=-0.420; P=0.040, r=-0.366). In squamous cell carcinoma patients, V(e) (homogeneity) was significantly negatively correlated with the expression of Ki-67 (P=0.033, r=-0.570). K(trans)(homogeneity) and V(e) (homogeneity, Q5, Q10, Q25) were significantly positively correlated with ADC values (P value from 0.001 to 0.035, r value from 0.545 to 0.765). Conclusion: DCE-MRI quantitative perfusion histogram parameters, ADC value can evaluate the lung cancer cell proliferation activity in different pathological types.

Low shear stress induces ERK nuclear localization and YAP activation to control the proliferation of breast cancer cells

Micro-environmental regulation of cancer cell malignancy is one of the most basic cancer life phenomena. However, the study of cellular response to microenvironment has been long focused on signal processes mediated by various chemical factors and their receptors, the study of mechanical forces, another key environmental factor, has been less studied. In recent years, more and more attention has been paid to the physiological and pathological significance of mechanical microenvironment. However, it is still not clear how cells perceive environmental changes and the signal pathways that regulate cell physiological activities. In this study, we identified that low shear stress (LSS) significantly promoted breast cancer cell proliferation. The proliferation was closely associated with mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase (ERK) and Yes-associated protein (YAP). Inhibition of ERK or YAP both abolished the LSS-induced proliferation activity of cancer cells. LSS induced ERK phosphorylation and YAP activations, which suggested the involvement of ERK and YAP under LSS treatment. Under LSS, ERK inhibitor U0126 decreased both active YAP and ERK expressions, while YAP inhibitor verteporfin failed to decrease ERK phosphorylation. Further study confirmed that ERK translocated to nucleus which showed an active state of ERK in LSS-treated group. LSS with verteporfin group showed no differences with LSS-treated group which confirmed ERK and YAP an upstream-downstream cascade. The above results demonstrated that LSS can promote breast cancer cell proliferation through ERK-YAP activation. These results not only highlight a new means of understanding mechanical transmission to cytoplasm mechanisms but also serve as a new basis for developing drug delivery systems for breast cancer treatment.

Expressions of Topo IIα and Ki67 in breast cancer and its clinicopathologic features and prognosis.

Objective:Breast cancer is one of the most common malignant tumors in women, and its incidence in women has ranked the first place among all malignant cancers. The evolution of breast cancer which is highly heterogeneous is a complex biological process involving multiple factors, genes and stages. Therefore, studying the expression and interaction of related genes is of great significance to further revealing of the mechanism of the occurrence and development of breast cancer and reduction of its resistance to chemotherapy. The purpose of this study was to investigate expressions of Ki67 and Topoisomerase IIα (Topo IIα) in breast cancer tissues and to explore the relationship between expressions of Ki67 and Topo IIα and the pathological characteristics and prognosis of breast cancer.Methods:Tissues with pathological changes and peripheral normal breast tissues were collected from 66 patients who underwent breast cancer treatment. Ki67 and Topo IIα in the breast cancer tissues and normal breast tissues were detected using immunohistochemical method. The relationships of Ki67 and Topo IIα with general clinicopathologic features and prognosis were analyzed.Results:It was found that the positive expressions of Ki67 and Topo IIα in the breast cancer tissues were higher than those in the normal tissues; the positive expression rate of Ki67 was in a correlation with diameter of breast cancer and lymphatic metastasis (P<0.05); the positive expression rate of Topo IIα was in a correlation with clinical stage, diameter of cancer and lymphatic metastasis (P<0.05). Multi-factor logistic regression analysis suggested that lymphatic metastasis and clinical stage were the independent predictive factors for positive expression of Topo IIα and lymphatic metastasis was the independent predictive factor for positive expression of Ki-67. Breast cancer patients with negative Ki67 and Topo IIα had significantly higher local recurrence rate than those who had positive Ki67 and Topo IIα (P<0.05) after surgery. The median time to progression (TTP) of the patients with positive expression of Topo IIα was lower than that with negative expression (P<0.05). The median TTP of the patients with negative and positive expression of Ki67 had no significant difference (P>0.05).Conclusion:Topo IIα and Ki67 can be used for reflecting the proliferation activity of cancer cells and can affect the postoperative recurrence and survival time after surgery. They may be involved in the occurrence, development and metastasis of breast cancer. They are expected to be indicators in the prognosis prediction of breast cancer and potential treatment targets.

PFKFB3促进腺样囊性癌ACC-2细胞增殖、迁移、侵袭和转移

Objective To evaluate the expression of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) in adenoid cystic carcinoma and the effects of PFKFB3 on the proliferation, migration, invasion and tumorigenesis in vivo of adenoid cystic carcinoma cells. Methods Immunohistochemistry and Western blotting were applied to detect the expression of PFKFB3 in 29 cases of adenoid cystic carcinoma tissues and 10 cases of para-carcinoma normal salivary gland tissues. PFKFB3 in adenoid cystic carcinoma ACC-2 cells was silenced by adenovirus vector. The effects of PFKFB3 silence on cell proliferation, migration, invasion and tumorigenesis in vivo and distant metastasis of adenoid cystic carcinoma cells were evaluated by cell proliferation assay, wound healing assay, Transwell assay, xenografted mice model and lung metastasis mice model. Results The results of immunohistochemical staining showed that the positive expression rate of PFKFB3 in adenoid cystic carcinoma tissues was 93.1% (27/29), and the positive expression rate of PFKFB3 in normal salivary gland tissues was 20.0% (2/10), with a significant difference (χ2=20.84, P<0.001). The results of methyl thiazolyl tetrazolium (MTT) assay showed that, compared with ACC-2 group, the proliferation activity of cancer cells with silence of PFKFB3 (PFKFB3--ACC-2) was significantly suppressed for 72 h (1.8±0.2 vs. 4.7±0.8, t=3.582, P=0.001). The results of wound healing assay showed that after scraping cells away for 24 h, the number of cells in the scratch area was 99.8±13.2 in the PFKFB3--ACC-2 group, which was significantly less than that in the ACC-2 group (263.0±97.4, t=2.868, P=0.029). Transwell results indicated that the number of cells passing through matrigel was 17.6±2.1 in the PFKFB3--ACC-2 group, which was less than that in the ACC-2 group (28.6±3.8, t=4.452, P=0.004). The tumor volume in the PFKFB3--ACC-2 [(623.5±134.1) mm3] was smaller than that in the ACC-2 group [(1 621.0±278.1) mm3,t=4.213, P=0.001]. More pulmonary metastases were found in the ACC-2 group than PFKFB3--ACC-2 group (18.1±3.2 vs. 4.1±2.2, t=6.322, P=0.001). Conclusion The expression of PFKFB3 is higher in adenoid cystic carcinoma tissue than normal salivary gland tissue, and the highly expressed PFKFB3 plays a driving role in the proliferation, migration, invasion and metastasis in adenoid cystic carcinoma. Key words: Carcinoma, adenoid cystic; Glycolysis; PFKFB3; Tumor development

The Importance of CYP19A1 in Estrogen Receptor-Positive Cholangiocarcinoma.

CYP19A1, also called aromatase, is a key enzyme for converting androgens to estrogens of estrogen synthesis. Elevated serum estrogen and high expression levels of estrogen-related proteins are found in cholangiocarcinoma (CCA; bile duct cancer). However, the expression of CYP19A1 in relation to estrogen-related proteins, including estrogen receptors (ERα, ERβ, and GPR30) and an estrogen response protein (TFF1), has never been explored in CCA. In this study, we investigated the expressions of CYP19A1 and estrogen-related proteins in CCA tissues (n = 74; 51 males and 23 females) using immunohistochemistry. The results showed that CYP19A1 was overexpressed in CCA cells compared with that in normal bile duct cells in the adjacent tissues. High expression of CYP19A1 was correlated with the metastatic status of the patients. High CYP19A1 expression was also positively correlated with GPR30 expression. Correlation between high CYP19A1 expression in the tumor tissues and shorter survival time was more prominent in male than in female CCA patients. To elucidate further, the effect of CYP19A1 knockdown on a CCA cell line was examined using a specific siRNA. When CYP19A1 gene expression was suppressed, migration and proliferation activities of CCA cells were significantly reduced. Moreover, the cell proliferation of high CYP19A1-expressing KKU-213 cells was more profoundly suppressed by CYP19A1 inhibitors (exemestane and letrozole) than low CYP19A1-expressing KKU-100 cells. Thus, CYP19A1 promotes CCA progression with aggressive clinical outcomes via increased migration and proliferation activities of cancer cells. CYP19A1 can be a potential chemotherapeutic target for CCA, especially in male patients.

Strategies and Methods for the Synthesis of Anticancer Natural Product Neopeltolide and its Analogs.

Neopeltolide, isolated in 2007, with its novel structural features and potent anti cancer cell proliferation activity, has attracted a tremendous amount of synthetic efforts. This review briefly and chronologically summarizes each of the synthesis with the main focus on the strategies and methodologies for the construction of its cis-tetrahydropyran-containing macrolactone core.

Abstract 1996: Inhibitor of differentiation-1 (Id1) expression deficiency in the tumor microenvironment impairs experimental hepatic metastasis of lung cancer

Abstract Background: Liver metastasis (LM) occurs in 30% of non-small cell lung cancer patients but the contribution of the patient's genetic background to the hepatic metastasis development is unclear. Previously, we reported that inflammation contributes to the prometastatic microenvironment of the liver. Upregulation of transcriptional regulator Id1 gene has been associated with inflammation while ablation of Id1 in mice reduced inflammation. In this study Id1 deficient mice were used to analyze the role of host Id1 in the hepatic colonization of an experimental lung cancer. Methods: Lewis lung carcinoma cells were used for the experimental production of hepatic metastasis via intrasplenic injection of cancer cells in C57BL/6 wild-type and Id1-knockout (KO) mice. Animals were splenectomized to avoid flank tumor formation and weekly FDG-micro-positron emission tomographies (PET) were performed to monitor LM formation. Animals were sacrificed at the time of LM occurrence and total RNA was purified from LM. A microarray gene expression analysis (Affymetrix) with the support of Ingenuity Pathways Analysis (IPA) was performed to evaluate the potential impact of Id1 deficiency on the regulation of genes mediating cancer cell invasion and proliferation, angiogenesis and metastasis. Results: By week two after cancer cell injection, 70% wild-type and 10% Id1-KO mice had detectable hepatic metastasis by FDG-PET (p=0.02). Three weeks after injection, when 100% wild-type had LM, still just 20% Id1-KO mice had LM (p=0.015). Moreover, 50% Id1-KO mice remained LM-free &amp;gt;4 weeks after cancer cell injection. No other metastasis sites were detected at necropsy. A microarray gene expression analysis of LM from Id1-KO animals uncovered a remarkable downregulation (p&amp;lt;0.05) of specific genes involved in the activation of cancer cell proliferation (Myc, Cdc20, Smc2, Aurora kinase B, Cyclin B1, CDK1, TIMP1, Epiregulin), migration (Ccl7, Serpine1/PAI-1, VIM, Anxa2, S100A4, S100A6, Akt3, Adrenomedullin), angiogenesis (Hif1a, PGF, Nestin) and metastasis (FGFR1, Src, IL-18, MMP3, MMP12, MMP13, Amphiregulin, PDGFA, FoxM1, Hsp90AA1, MIF, Ccl2, RhoC). Conclusion: Our results demonstrate that Id1 expression deficiency impaired the metastatic process of lung cancer cells to the liver through the specific downregulation of key metastasis-associated genes and suggest that Id1-dependent mechanisms are new targets for hepatic metastasis therapeutic. This study has been partially funded by “UTE project CIMA” and an ISCIII-FIS grant 2011. Citation Format: Ignacio Gil-Bazo, Eduardo Castanon, Inés López, Victor Segura, Mariano Ponz-Sarvise, José M. López-Picazo, Maria Collantes, Margarita Ecay, Isabel Gil-Aldea, Alfonso Calvo, Fernando Vidal-Vanaclocha. Inhibitor of differentiation-1 (Id1) expression deficiency in the tumor microenvironment impairs experimental hepatic metastasis of lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1996. doi:10.1158/1538-7445.AM2014-1996

Expression and clinical significance of PCNA and IRF2 in human pancreatic cancer

Objective To research the expression levels,clinical significance and the correlation of PCNA and IRF2 in human pancreatic cancer.Methods The expression levels of PCNA and IRF2 in 60 different cases of pancreatic cancer tissues and 30 different cases of para-pancreatic cancer tissues were investigated by Immunohistochemical SP staining and then analyzed through clinicopathological data.Results Immunohistochemical results:the positive expression rates of PCNA and IRF2 in pancreatic cancer tissues were 83.3％ and 78.3％ respectively,which were significantly higher than those in the adjacent pancreatic tissues,of which were 20.0％ and 23.3％ respectively.The differences were statistically significant (P＜0.001).The factors of age,gender,tumor site of patients with pancreatic carcinoma were not statistically significant with the expression of PCNA (P＞0.05),but other factors,such as the differentiation of pancreatic tissue,TNM stage and lymph node metastasis,were statistically significant (P＜O.05).The factors of age,sex,tumor location and lymph node metastasis of pancreatic carcinoma patients were not statistically significant with the expression of PCNA (P＞0.05),but the factors of differentiation of pancreatic tissue and TNM stage were statistically significant (P＜0.05).Moreover,the expression of PCNA and IRF2 was positively correlated (γ =0.416,P=0.001).Conclusion The abnormal expression of PCNA and IRF2 is correlated with occurrence of pancreatic cancer and the cancer cell proliferation activity. Key words: Pancreatic Cancer; PCNA; IRF2; Immunohistochemistry

Effects of dendritic cell-cytokine-induced killer cells on proliferation and telomerase activity of human colon cancer cells

Objective To explore the effects and mechanism of cytokine induced killer (CIK)cells and dendritic cells (DC) from peripheral blood mononuclear cells (PBMC) of patients with human colorectal cancer on prloferation and apoptosis of human liver cancer cell line SW620.Methods Adherent PBMC were induced by granulocyte macrophage-colony stimulating factor (GM-CSF),tumor necrosis factor (TNF)-α and interleukin (IL)-4 to prepare DC.CIK cells were obtained from suspended PBMC induced by interferon (IFN)-γ,IL-2 and human CD3 monoclonal antibody.The colorectal cancer SW620 cells that were cultured without DC-CIK cells served as the controls.DC-CIK cells and SW620 cells were cultured in a well cell cultrue cluster for 24,48 and 72 h,respectively.The proliferation of SW620 cells that were cultured with DC-CIK cells for 24,48 and 72 h was detected by methyl thiazol tetrazolium (MTT) assay.The telomerase activity was determined by tartrate resistant acid phosphatase (TRAP)-enzyme linked immunosorbent assay (ELISA).Human telomerase reverse transcriptase (hTERT) gene was evaluated by Western blotting.Results In colorectal cancer cells treated with DC-CIK after 72 h treatment,cell relative growth rate in control group,and DC-CIK-a group and DC-CIK-b group was (95.6 ± 2.2) ％,(73.5 ±1.8)％ and (51.8 ± 1.6)％ respectively.The results from TRAP-ELISA showed that telomerase activity in control group at 24,48 and 72 h was 1.568 ±0.078,1.535 ±0.056,and 1.527 ±0.039,that in DCCIK-a group was 1.356 ±0.056,1.108 ±0.039,and 0.578 ±0.028,and that in DC-CIK-b was 1.181 ±0.075,0.718 ± 0.039,and 0.386 ± 0.036,respectively.The Western blotting results showed that the hTERT protein in DC-CIK-a group and DC-CIK-b group was down-regulated as compared with control group.Conclusion DC-CIK cells could inhibit the proliferation of colon cancer cells through down-regulating telomerase activity. Key words: Colorectal cancer; Dendritic cell-cytokine induced killer; Proliferation; Telomerase

The effects on proliferation and invasiveness activities of gastric cancer cells initiated by macrophages

Objective To observe the biological effects on human gastric cancer cell line AGS exerted by macrophages and discuss the role of tumor-associated macrophages (TAMs) on gastric cancer invasion and metastasis.Methods Human monocytic cell line THP-1 were differentiated to macrophages in vitro,then cocultured with AGS cells to mimic the micro-environment of gastric cancer.The effects on proliferation and invasiveness activities of gastric cancer cells initiated by macrophages were assessed.Results The AGS cells cocultured with macrophages were shown decreased proliferation,and the cell doubhng time was (38.42 ± 2.68) h,which was (24.36 ± 2.91) h in those non-cocuhured (P ＜ 0.001).In vitro invasion assay indicated that the invasive cell count of AGS cells cocultured with macrophages and those noncocultured was 308 ± 15 and 168 ±4 separately (P ＜0.01).The effects of macrophages on gastric cancer cells were related to the cytokines.The THP-1derived macrophages secreted interleukin (IL)-1β 20 fold higher (P ＜0.01),IL-8 40 fold higher (P ＜0.01) and tumor necrosis factor (TNF)-α 8 fold higher (P ＞0.05) than those THP-1 cells did.Conclusion macrophages could enhance the invasiveness of gastric cancer cells,whith may ralated to their inceased secretion of IL-1β,IL-8 and TNF-α. Key words: Tumor-associated macrophages ; Gastric cancer; Invasiveness

Effect of lactic fermentation on the total phenolic, saponin and phytic acid contents as well as anti-colon cancer cell proliferation activity of soymilk

In an attempt to develop healthy dietary adjuncts, soymilk was fermented simultaneously with Streptococcus thermophilus 14085 and Bifidobacterium infantis 14603 at 37°C for 24h. It was found that lactic fermentation reduced the content of saponins and phytates, which possess antinutritional activity, and enhanced the total phenolic content as well as antitumor cell proliferation effect of soymilk against HT-29 and Caco-2 cells. The original antitumor cell component, starter organisms, and antitumor cell bioactive principles formed in soymilk during fermentation, might all have contributed to the enhanced antitumor activity of fermented soymilk. The antiproliferative effect of the extracts varied with extraction solvent. Extracts obtained from fermented soymilk with 80% methanol exhibited the highest suppression effect on the proliferation of HT-29 and Caco-2 cells. This study further stresses the potential of developing soymilk as a healthy dietary adjunct possessing enhanced anticancer activity through the use of lactic fermentation.

ASSESSMENT OF WOUND HEALING ACTIVITY OF DAPHNE OLEOIDES SUBSP. KURDICA: IDENTIFICATION OF THE ACTIVE COMPONENTS USING BIOASSAY GUIDED ISOLATION METHOD

s. PHYTOPHARM 2012 M 76 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 CRS) and aspirin (4h ASP) induced ulceration. The ulcer index and % protection was calculated. The gastric juice collected from 4h Pl rats was studied for the effects on gastric volume, ph, free acid, total acid-pepsin output, dNA gastric juice cell shedding and mucosal cell proliferation. biochemical estimation such as histamine content, mucus content and hkATPase inhibitory activity was also evaluated. The effect of APf on stomach histopathology was also studied. Statistical analysis was done by one way ANOVA followed by dunnett’s post hoc test for multiple comparisons against control group. The difference was considered to be significant when P < 0.05. Oral treatment of APf (200 and 400 mg/kg, p. o.) for 7 days showed significant protection against acute gastric ulcer induced by 4h Pl and 95 % EtOh, but was found to have no effect on CRS and ASP-induced gastric ulceration. The gastric studies showed that treatment with APf signiccantly increased the gastric juice ph, decrease the gastric volume and total acid-pepsin output and significantly reduced the gastric juice cell shedding. A significant decrease in histamine content and a reduction in free phosphate ions responsible for hkATPase inhibitory activity were also observed. in conclusion, the results of the present study indicate that the gastroprotective activity of APf contributes mainly to the peripheral effects on the gastric secretion and the probable mechanism underlying behind such activity might be both due to the possible inhibition on the histaminic (h 2 ) receptor and hkATPase pump. INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY © Khoo Gaik Ming, Clausen Morten Rahr, Larsen Erik Department of Food Science, Aarhus University, Kirstinebjergvej 10, DK-5792 Aarslev, Denmark The object of this study was to investigate the inhibitory effect of betulinic acid, oleanolic acid and ursolic acid on human colon cancer cell proliferation activity. Triterpene acids are members of the terpene family and are widely distributed in food plants. A rosehip fraction containing a mixture of betulinic acid, oleanolic acid and ursolic acid displayed inhibitory effect against the release of pro-inflammatory cytokine interleukin-6 with the combination of oleanolic acid/betulinic acid and ursolic acid/betulinic acid displaying significant synergistic inhibitory effects (1). Our results showed that these three compounds also displayed good inhibitory effect against cancer cell proliferation activity at the concentration of 10μg/ml. both betulinic acid and ursolic acid displayed strong inhibition against hT-29 (88 % and 79 %, respectively) and SW-480 (76 % and 82 %, respectively), but a moderate to weak inhibition against Caco-2 (19 % and 22 %, respectively). Oleanolic acid displayed only moderate inhibition against Caco-2 (49 %), very weak inhibition against hT-29 (7 %) and no inhibition against SW-480. No synergistic effects were observed between combinations of the triterpene acids. We only found an additive effect in the combination of oleanolic acid/ursolic acid and only against proliferation of hT-29. Antagonistic effects between the pure compounds were observed when betulinic acid was one of the components in the combination. Although betulinic acid and ursolic acid showed good inhibitory effects against all three cancer cell lines, their combination displayed antagonistic effects and showed poor inhibition against cancer cell proliferations. however, these triterpenic acids and their combination effects should be further investigated in different in vitro or in vivo experiment to confirm their bioactivities and mechanism. Reference: (1) Saaby l., Moesby l., hansen E. W., Christensen S. b. (2011). isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina l.). Phytotherapy Research, 25, 195–201. DECURSIN AND RELATED COMPOUNDS INHIBIT VASCULAR ENDOTHELIAL MIGRATION BY SPHINGOSINE KINASE INHIBITION © Lee Yong-Moon, Shin Kyong-Oh, Seo Cho-Hee, Kim Tack-Joong College of Pharmacy, Chungbuk National University, Cheongju, South Korea Division of Biological Science and Technology, Yonsei University, Wonju, South Korea The pyranocoumarin compound decursin isolated from the herb, Angelica gigas, are known to possess potent anti-inflammatory activities. however, little is known about their anti-angiogenic activity. here, we investigated the anti-angiogenic effects of decursin by the changes of sphingolipid metabolites

INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY

s. PHYTOPHARM 2012 M 76 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 CRS) and aspirin (4h ASP) induced ulceration. The ulcer index and % protection was calculated. The gastric juice collected from 4h Pl rats was studied for the effects on gastric volume, ph, free acid, total acid-pepsin output, dNA gastric juice cell shedding and mucosal cell proliferation. biochemical estimation such as histamine content, mucus content and hkATPase inhibitory activity was also evaluated. The effect of APf on stomach histopathology was also studied. Statistical analysis was done by one way ANOVA followed by dunnett’s post hoc test for multiple comparisons against control group. The difference was considered to be significant when P < 0.05. Oral treatment of APf (200 and 400 mg/kg, p. o.) for 7 days showed significant protection against acute gastric ulcer induced by 4h Pl and 95 % EtOh, but was found to have no effect on CRS and ASP-induced gastric ulceration. The gastric studies showed that treatment with APf signiccantly increased the gastric juice ph, decrease the gastric volume and total acid-pepsin output and significantly reduced the gastric juice cell shedding. A significant decrease in histamine content and a reduction in free phosphate ions responsible for hkATPase inhibitory activity were also observed. in conclusion, the results of the present study indicate that the gastroprotective activity of APf contributes mainly to the peripheral effects on the gastric secretion and the probable mechanism underlying behind such activity might be both due to the possible inhibition on the histaminic (h 2 ) receptor and hkATPase pump. INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY © Khoo Gaik Ming, Clausen Morten Rahr, Larsen Erik Department of Food Science, Aarhus University, Kirstinebjergvej 10, DK-5792 Aarslev, Denmark The object of this study was to investigate the inhibitory effect of betulinic acid, oleanolic acid and ursolic acid on human colon cancer cell proliferation activity. Triterpene acids are members of the terpene family and are widely distributed in food plants. A rosehip fraction containing a mixture of betulinic acid, oleanolic acid and ursolic acid displayed inhibitory effect against the release of pro-inflammatory cytokine interleukin-6 with the combination of oleanolic acid/betulinic acid and ursolic acid/betulinic acid displaying significant synergistic inhibitory effects (1). Our results showed that these three compounds also displayed good inhibitory effect against cancer cell proliferation activity at the concentration of 10μg/ml. both betulinic acid and ursolic acid displayed strong inhibition against hT-29 (88 % and 79 %, respectively) and SW-480 (76 % and 82 %, respectively), but a moderate to weak inhibition against Caco-2 (19 % and 22 %, respectively). Oleanolic acid displayed only moderate inhibition against Caco-2 (49 %), very weak inhibition against hT-29 (7 %) and no inhibition against SW-480. No synergistic effects were observed between combinations of the triterpene acids. We only found an additive effect in the combination of oleanolic acid/ursolic acid and only against proliferation of hT-29. Antagonistic effects between the pure compounds were observed when betulinic acid was one of the components in the combination. Although betulinic acid and ursolic acid showed good inhibitory effects against all three cancer cell lines, their combination displayed antagonistic effects and showed poor inhibition against cancer cell proliferations. however, these triterpenic acids and their combination effects should be further investigated in different in vitro or in vivo experiment to confirm their bioactivities and mechanism. Reference: (1) Saaby l., Moesby l., hansen E. W., Christensen S. b. (2011). isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina l.). Phytotherapy Research, 25, 195–201. DECURSIN AND RELATED COMPOUNDS INHIBIT VASCULAR ENDOTHELIAL MIGRATION BY SPHINGOSINE KINASE INHIBITION © Lee Yong-Moon, Shin Kyong-Oh, Seo Cho-Hee, Kim Tack-Joong College of Pharmacy, Chungbuk National University, Cheongju, South Korea Division of Biological Science and Technology, Yonsei University, Wonju, South Korea The pyranocoumarin compound decursin isolated from the herb, Angelica gigas, are known to possess potent anti-inflammatory activities. however, little is known about their anti-angiogenic activity. here, we investigated the anti-angiogenic effects of decursin by the changes of sphingolipid metabolites

Abstract B13: Cycloartane-3,24,25-triol: A cancer chemopreventive cycloartane inhibits MRCKα kinase and demonstrates anti prostate cancer activity in vitro

Abstract In our study of the Jamaican ball moss (Tillandsia recurvata L.) for its prostate cancer chemotherapeutic properties, we have identified a number of cycloartane-type compounds from the chloroform crude extract. Cycloart-23-ene-3,25-diol was isolated and characterized from the extract. Attempts to run this compound in our kinase and antiproliferative assays were unsuccessful because of solubility problems. A search of the literature led us to Cycloartane-3,24,25-triol, a close analog of Cycloart-23-ene-3,25-diol. Cycloart-23-ene-3,25-diol differs from Cycloartane-3,24,25-triol only in the presence of a third hydroxyl group in this compound while Cycloart-23-ene-3,25-diol has only two hydroxyl groups. We isolated Cycloartane-3,24,25-triol from its source plant (Chrysanthemum morifolium) and screened it against a 451 kinase panel for inhibitory properties as well as for anti- prostate cancer cell proliferation activity using DU145 and PC-3 cell lines. Cycloartane-3,24,25-triol inhibited MRCKα (Myotonic dystrophy kinase-related Cdc42–binding kinase) kinase at a Kd of 0.26μM and showed IC50 values of 1.56±0.18μM and 2.04±0.28μM against DU145 and PC-3 prostate cancer cell lines respectively. MRCK kinases are now known to contribute to the process of tumor progression and metastasis through the control of the cytoskeleton which regulates tumor cell motility and invasion. In prostate cancer, the MRCKα kinase indirectly contributes to tumor progression by activating other kinases including the Lim kinase 1 (LIMK 1) which is over-expressed in prostate cancer and known to contribute to tumor cell invasion and migration leading to metastasis to other organs. This is the first report on the anti-prostate cancer activity and kinase inhibitory activity of Cycloartane-3,24,25-triol. The combined kinase and in vitro antiproliferation activity of Cycloartane-3,24,25-triol makes it a good candidate for further studies to determine its in vivo efficacy against prostate cancer. Citation Format: Henry I.C. Lowe, Ngeh J. Toyang, Joseph Bryant. Cycloartane-3,24,25-triol: A cancer chemopreventive cycloartane inhibits MRCKα kinase and demonstrates anti prostate cancer activity in vitro [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B13.

HSulf-1 Gene Exhibits Anticancer Efficacy through Negatively Regulating VEGFR-2 Signaling in Human Cancers

BackgroundHuman sulfatase 1 (hSulf-1) is a heparin-degrading endosulfatase that desulfates cell surface heparan sulfate proteoglycans (HSPGs) in extracellular matrix and negatively modulates heparin-binding growth factor and cytokine signaling in cell proliferation. But hSulf-1 function is more complicated, and its molecular mechanism has not been well known.Principal FindingsTo further investigate the functions of hSulf-1 gene in regulating the vascular endothelial growth factor receptor (VEGFR) signaling, a series of vectors expressing hSulf-1, hSulf-1 small hairpin RNA (shRNA) and VEGFR-2 shRNA were generated. hSulf-1 re-expression could downregualte the VEGFR-2 phosphorylation and inhibit cancer cell proliferation both in ovarian and hepatocellular cancer cell lines. Knockdown of hSulf-1 expression by hSulf-1 shRNA enhanced the recovery of high levels of phosphorylated VEGFR-2, and knockdown of VEGFR-2 expression by VEGFR-2 shRNA inhibited the proliferation activity of cancer cells in vitro to some extent. In human cancer xenografts in nude mice, tumor growth was inhibited markedly after injections of adenovirus expressing hSulf-1, with the tumor inhibition rates of 46.19% and 49.56% in ovarian and hepatocellular tumor models, respectively. hSulf-1 expression significantly reduced tumor microvessel density.ConclusionsThe results demonstrated that hSulf-1 re-expression both in ovarian and hepatocellular cancer cells induces antitumor efficacy by attenuating the phosphorylation of VEGFR-2 and suppressing angiogenesis. Therefore, hSulf-1-mediated antiproliferation and antiangiogenesis could be a reasonable approach for cancer therapy.

Construction of the antisense eukaryotic vector for proliferating cell nuclear antigen gene and its expression in bladder cancer EJ cell line.

To explore a novel strategy for antisense gene therapy of cancer, the coding sequence of human proliferating cell nuclear antigen (PCNA) cDNA was reversely inserted into the eukaryotic vector pLXSN by molecular cloning techniques and transferred into bladder cancer EJ cells with liposome. The PCNA expression in transferred cells was dynamically detected by immunofluorescence and RT-PCR techniques. Changes of proliferation activities of cancer cells were assayed by MTT colorimetric and cloning formation methods. In the experiment, the antisense eukaryotic vector was successfully constructed and named as pLAPSN. After transfection with it for 1-7 days, PCNA protein and mRNA levels in cancer cells were blocked by 16.74%-84.21% (P < 0.05) and 23.27%-86.15% (P < 0.05) respectively. The proliferation activities of transferred cells were inhibited by 27.91%-62.07% (P < 0.01), with cloning formation abilities being decreased by 50.81% (P < 0.01). It was concluded that the in vitro proliferation activities of cancer cells could be effectively inhibited by blocking PCNA expression with antisense technique, which could serve as an ideal strategy for gene therapy of bladder cancer.

Is a proliferation index of cancer cells a reliable prognostic factor after hepatectomy in patients with colorectal liver metastases?

Background: In spite of many reports focusing on prognostic factors after hepatectomy in patients with colorectal liver metastases, few studies have investigated pathological factors, eg, fibrous pseudocapsulation, growth pattern at the tumor margin, and proliferation activity of cancer cells, other than histological type and surgical margin. The aim of the present study was to investigate whether absence of pseudocapsulation, infiltrative growth pattern of metastases, and higher proliferation of cancer cells shown by Ki-67 immunohistochemical reactivity were associated with poorer survival after hepatectomy among patients with colorectal liver metastases. Methods: Between 1988 and 1998, 221 patients underwent hepatic resection of colorectal metastases with curative intent in our institution. Pathology analyses were focused on pseudocapsulation of liver metastases, growth pattern at the tumor edge, and Ki-67 labelling index (Ki-67 LI) of cancer cell nuclei. Univariate analyses of survival and of disease-free survival were performed for several clinicopathological factors, and multivariate analyses of survival and disease-free survival were also performed. Results: The univariate survival analyses showed that pseudocapsulation, growth pattern, and Ki-67 LI were significant prognostic factors, besides synchronous versus metachronous occurrence of metastases, carcinoembryonic antigen level before hepatectomy, and number of metastases. A multivariate analysis showed that Ki-67 labeling index was the most reliable prognostic factor of survival. In addition, Ki-67 LI and microscopic growth pattern were multivariately predictive factors of disease-free survival. Conclusions: This large single-institution study showed that investigation of cancer cell proliferation and pathologic characteristics of the tumor margin are major prognostic factors.