Inhibition Rate Of Tumor Weight Research Articles

TPGS nanoparticles co-loaded with ABT-737 and R848 for breast cancer therapy

The development of new effective drugs to treat breast cancer remains a huge challenge. ABT-737 can inhibit Bcl-2 proteins to promote apoptosis. Resiquimod (R848) is a TLR7/8 agonist that is effective in modulating the immunosuppressive microenvironment. In this study, a codelivery system (TPGS/ABT+R848 NPs) based on D-α-tocopheryl poly (ethylene glycol) 1000 succinate as a potential drug delivery vector to codelivery ABT-737 and R848 was investigated. The size of TPGS/ABT+R848 NPs was 102.5 nm, the drug loading of ABT-737 and R848 was 30.6 % and 12.5 %, and the entrapment efficiency was 84.2 % and 23.7 %, respectively. The nanoparticles showed no significant change in particle size over 14 days. R848 and ABT-737 were released in co-loaded nanoparticles in sequential order. In vitro anti-tumor experiments, the IC50 value of TPGS/ABT+R848 NPs was 0.30 μg·mL−1, 34 times lower than that of free ABT-737. Animal experiments also verified that TPGS/ABT+R848 NPs could enhance the anti-tumor activity, and the tumor weight inhibition rate was 75.3 %. This study demonstrated that TPGS NPs loaded with ABT-737 and R848 have superior combination tumor therapeutic effects, and the co-loaded preparation is conducive to anti-tumor efficacy. The TPGS/ABT+R848 NPs could be a promising platform against breast cancer.

Programing Immunogenic Cell Death in Breast Tumors with Designer DNA Frameworks.

The low response rate and serious side effects of cancer treatment pose significant limitations in immunotherapy. Here, we developed a multifunctional tetrahedral DNA framework (TDF) as a drug carrier to recruit chemotherapeutants and trigger immunogenic cell death (ICD) effects, which could turn tumors from cold to hot to boost the efficacy of antitumor immunotherapy. A tumor-targeting peptide RGD was modified on the TDF to increase the delivery efficiency, and the chemotherapeutant doxorubicin (DOX) was loaded to induce ICD effects, which were assisted by the immune adjuvant of CpG immunologic sequences linked on TDF. We demonstrated that the multifunctional TDF could suppress 4T1 breast tumor growth by increasing tumor infiltration of CD8+ T cells, upregulating granzyme B and perforin expressions to twice as much as the control group, and decreasing 30% CD25+ Treg cells. Furthermore, the combination of α-PD-1 could inhibit the growth of distant tumor and suppressed tumor recurrence in a bilateral syngeneic 4T1 mouse model; the distant tumor weight inhibition rate was about 91.6%. Hence, through quantitatively targeting the delivery of DOX to reduce the side effects of chemotherapy and sensitizing the immune response by ICD effects, this multifunctional TDF therapeutic strategy displayed better treatment effect and a promising clinical application prospect.

The anti-tumor and renoprotection study of E-[c(RGDfK)2]/folic acid co-modified nanostructured lipid carrier loaded with doxorubicin hydrochloride/salvianolic acid A

BackgroundPoor in vivo targeting of tumors by chemotherapeutic drugs reduces their anti-cancer efficacy in the clinic. The discovery of over-expressed components on the tumor cell surface and their specific ligands provide a basis for targeting tumor cells. However, the differences in the expression levels of these receptors on the tumor cell surface limit the clinical application of anti-tumor preparations modified by a single ligand. Meanwhile, toxicity of chemotherapeutic drugs leads to poor tolerance to anti-tumor therapy. The discovery of natural active products antagonizing these toxic side effects offers an avenue for relieving cancer patients’ pain during the treatment process. Since the advent of nanotechnology, interventions, such as loading appropriate drug combinations into nano-sized carriers and multiple tumor-targeting functional modifications on the carrier surface to enhance the anti-tumor effect and reduce toxic and side effects, have been widely used for treating tumors.ResultsNanocarriers containing doxorubicin hydrochloride (DOX) and salvianolic acid A (Sal A) are spherical with a diameter of about 18 nm; the encapsulation efficiency of both DOX and salvianolic acid A is greater than 80%. E-[c(RGDfK)2]/folic acid (FA) co-modification enabled nanostructured lipid carriers (NLC) to efficiently target a variety of tumor cells, including 4T1, MDA-MB-231, MCF-7, and A549 cells in vitro. Compared with other preparations (Sal A solution, NLC-Sal A, DOX solution, DOX injection, Sal A/DOX solution, NLC-DOX, NLC-Sal A/DOX, and E-[c(RGDfK)2]/FA-NLC-Sal A/DOX) in this experiment, the prepared E-[c(RGDfK)2]/FA-NLC-Sal A/DOX had the best anti-tumor effect. Compared with the normal saline group, it had the highest tumor volume inhibition rate (90.72%), the highest tumor weight inhibition rate (83.94%), led to the highest proportion of apoptosis among the tumor cells (61.30%) and the lowest fluorescence intensity of proliferation among the tumor cells (0.0083 ± 0.0011). Moreover, E-[c(RGDfK)2]/FA-NLC-Sal A/DOX had a low level of nephrotoxicity, with a low creatinine (Cre) concentration of 52.58 μmoL/L in the blood of mice, and no abnormalities were seen on pathological examination of the isolated kidneys at the end of the study. Sal A can antagonize the nephrotoxic effect of DOX. Free Sal A reduced the Cre concentration of the free DOX group by 61.64%. In NLC groups, Sal A reduced the Cre concentration of the DOX group by 42.47%. The E-[c(RGDfK)2]/FA modification reduced the side effects of the drug on the kidney, and the Cre concentration was reduced by 46.35% compared with the NLC-Sal A/DOX group. These interventions can potentially improve the tolerance of cancer patients to chemotherapy.ConclusionThe E-[c(RGDfK)2]/FA co-modified DOX/Sal A multifunctional nano-drug delivery system has a good therapeutic effect on tumors and low nephrotoxicity and is a promising anti-cancer strategy.Graphical

Dual-RNA controlled delivery system inhibited tumor growth by apoptosis induction and TME activation

Drug-controlled release is recognized as effective for improving compliance with treatment and obtaining better therapeutic efficacy with less toxicity in cancer treatment. However, few reports in this area are involved in nucleic acids delivery, especially in RNA therapeutics delivery. In this study, an injectable hydrogel Methacrylated gelatin (GM) scaffold was introduced into a dual-RNA hybrid delivery complex hybrid lipid particle (HLP) to form a G-HLP/RNAs system. This system can control the release of both siRNA and mRNA and was found to be efficient for protecting these RNAs from biodegradation and retaining their therapeutic effect over 7 days. Further, a tumor environment (TME)-activation function after peritumoral injection of mocked GM scaffold was observed. Then, matured DC cells and activated T-cells were detected by the addition of HLP/RNAs complex, thus verifying the immunoactivation function of GM scaffold and its ability to reserve immune cells and antigens. Finally, two doses of G-HLP/RNAs treatment efficiently suppressed C26 tumor growth in mice with a tumor weight inhibition rate of 71.9%. Owing to its ability to achieve RNA drug-controlled release, alter TME, and induce tumor apoptosis, the G-HLP/RNAs system may become a valuable tool for cancer gene therapy.

Sorafenib Nanomicelles Effectively Shrink Tumors by Vaginal Administration for Preoperative Chemotherapy of Cervical Cancer.

To investigate the potential of sorafenib (SF) in preoperative chemotherapy for cervical cancer to reduce tumor volume, sorafenib micelles (SF micelles) with good stability and high drug loading were designed. SF micelles were prepared by film hydration followed by the ultrasonic method. The results showed that the SF micelles were spherical with an average particle size of 67.18 ± 0.66 nm (PDI 0.17 ± 0.01), a considerable drug loading of 15.9 ± 0.46% (w/w%) and satisfactory stability in buffers containing plasma or not for at least 2 days. In vitro release showed that SF was gradually released from SF micelles and almost completely released on the third day. The results of in vitro cellular intake, cytotoxicity and proliferation of cervical cancer cell TC-1 showed that SF micelles were superior to sorafenib (Free SF). For intravaginal administration, SF micelles were dispersed in HPMC (SF micelles/HPMC), showed good viscosity sustained-release profiles in vitro and exhibited extended residence in intravaginal in vivo. Compared with SF micelles dispersed in N.S. (SF micelles/N.S.), SF micelles/HPMC significantly reduced tumor size with a tumor weight inhibition rate of 73%. The results suggested that SF micelles had good potential for preoperative tumor shrinkage and improving the quality life of patients.

Abstract 2092: Effects of Yi Sui Granules, an herbal medicine, on MDS/AML: Similar to decitabine or other mechanisms

Abstract Objective Yi Sui Granules (YSG) has been clinically used and proved efficacious in patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) for 25 years, however, its mechanism remains unclear. This study is to evaluate the effects of YSG on DNA methylation of AML/MDS. Methods and Results From cytotoxicity assays using CCK-8 method, we found out YSG inhibited SKM-1, HL-60, K-562 and CEM cells in a dose-dependent manner with an IC50 of 1.173±0.165 mg/ml, 1.70±0.2 mg/ml, 2.85±0.05 mg/ml, 1.70±0.2 mg/ml respectively. In vivo, decitabine inhibited tumor growth in terms of tumor volume and tumor weight but led to significant weight loss of SKM-1 xenographed BALB/c-nu mice. While YSG (50.69%) showed similar tumor weight inhibition rate to decitabine (56.64%) without side effects including body weight loss. To explore its mechanism, we discovered that DNA methyltransferase 1 (DNMT1) expression was significantly down-regulated(P<0.05) and SFRP5 gene was demethylated(P<0.01) after treated with either YSG or decitabine using RT-PCR, western blot and bisulfite amplicon sequencing methods. Furthermore, we found significant up-regulation of SFRP5 expression(P<0.05), down-regulation of Wnt3a protein(P<0.05) and β-catenin expression(P<0.05), as well as down-regulation of C-myc(P<0.05) and CyclinD1 expression(P<0.05) in decitabine group and YSG group. Equally interestingly, survival benefits were observed in YSG high-dose group but not in Decitabine group(P<0.05). Conclusion Similar to decitabine, YSG's suppression of SKM1 tumor may be related to the inhibition of DNMT1 expression, de-methylation of SFRP5 and up-regulation of SFRP5 expression, preventing Wnt3a from binding to Frizzled receptors, therefore inhibiting the abnormal activation of Wnt signaling, and down-regulating its target gene expression including C-myc and CyclinD1. Unlike decitabine, YSG prolong overall survival though other mechanisms, which will be further investigated in our future research. Corresponding Author: Jing Wang and Xinyi Chen, Address: No.5 Haiyuncang, Dongcheng District, Beijing, 100700, China; Email: Jing.Wang@bucm.edu.cn and chenxinyi0729@126.comAcknowledgement: This work was supported by the Young Scientist Fund (81503575) and Key Program (81630080) of National Natural Science Foundation of China, the Golden Bridge Project of Beijing Association for Science and Technology(ZZ20059) and the Elite Program of Dongcheng district of Beijing (2020-dchrcpyzz-38). Citation Format: Jing Wang, Chong Gao, Elizabeth Gullen, Jieya Wu, Xiaoyuan Zhang, Xinyi Chen. Effects of Yi Sui Granules, an herbal medicine, on MDS/AML: Similar to decitabine or other mechanisms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2092.

Chitosan oligosaccharides with degree of polymerization 2–6 induces apoptosis in human colon carcinoma HCT116 cells

Colon cancer is the third most common cancer, and yet there is a lack of effective therapeutic method with low side effects. Chitosan oligosaccharides (COS) is derived from chitosan after chitin deacetylation, and attracts more interests due to smaller molecular weight and soluble property. Previously, COS, mainly absorbed through intestinal epithelia, has been reported to exhibit many bioactivities, especially its anti-tumor effect. Recent references pay little attention to molecular weight distribution which is crucial for understanding its biological behavior. Here, we studied reducing sugar content and degree of polymerization (DP) of COS. 86.73% reducing sugar exists in COS sample and the content of chitosan fractions with 2–6 is 85.8%. COS suppressed the growth of HCT116 cells in vitro and in vivo, and the inhibition rate of tumor weight in vivo was high up to 58.6%. Moreover, the morphology observation, flow cytometry analysis and mRNA expression were applied to study the apoptosis related mechanism. COS treatment promoted mitosis, late stage apoptosis and S cell cycle arrest in HCT116 cells, and enhanced the mRNA expression of BAK and reduce BCL-2 and BCL-xL. These findings may provide an important clue for clinical applications of COS as anti-tumor drug or pharmaceutic adjuvant in the future.

Synergistic Inhibitory Effect of Arsenic Trioxide Combined with Itraconazole on Hedgehog Pathway of Multiple Myeloma NCI-H929 Cells

To investigate the antitumor effect of arsenic trioxide (ATO) combined with itraconazole (ITRA) on human multiple myeloma NCI-H929 cells by synergistically inhibiting Hedgehog (HH) signaling pathway. The inhibitory rate of NCI-H929 cells was assayed by MTT method. Tumor weight, tumor weight inhibition rate, and tumor volume of mouse model with multiple myeloma were examined. The ELISA were appled to detect the M-protein, qPCR and Western blot were used respectively to detect the expression level of Ptch, SMO, Gli and downstream target genes, the survival rate of tumor-bearing mice was analyzed. ATO combined with ITRA significantly inhibited NCI-H929 cell proliferation as compared with a single administration. The combination of ATO and ITRA could synergistically inhibit tumor growth and obviously reduced tumor burden, survival time of tumor-bearing mice was significantly prolonged. qPCR and Western blot results confirmed that the ATO combined with ITRA could significantly down-regulated expression of Gli1, leading to significantly decrease of cyclinD1 and BCL-2 expression levels. ATO combined with ITRA can more strongly suppress the growth of multiple myeloma NCI-H929 cells, as compared with a single administration. The synergistic effect of ATO and ITRA significantly down-regulates expression of Gli1 in HH signaling pathway, moreover the inhibition of target gene overexpression may be one of two drug mechanisms.

Inhibitory effect and its mechanism of CS055 on human colon carcinoma transplanted tumor

Objective To explore the inhibitory effect and its mechanism of CS055[a selective histone deacetylase inhibitor(HDACi)]on human colon carcinoma in nude mice. Methods The xenograft model was established successfully.Fifty-five mice were divided into five groups and eleven blocks by randomized complete-block design.The mice were respectively gavaged by CS055 10. 0 mg/(kg·d)in the first group,5 mg/ (kg·d)in the second group and 2. 5 mg/(kg·d)in the third group, while the mice in fourth group were treated by intraperitoneal injection of 5-fluorouracil in a dose of 25. 0 mg/(kg·d)and those in the fifth group were gavaged by negative-control-drug 0. 1 ml/(10 g·d)as negative control group.After treatment, all nude mice were killed,and all the subcutaneous transplanted tumors were stripped and measured by calipers and scales.Inhibitory rate of tumor weight and tumor volume was calculated.Tissue hematoxylin and eosin (HE)staining and transmission electron microscopy detection were used to detect the tumor cell apoptosis. Western blotting was used to detect the expression of histone deacetylase 1(HDAC1), HDAC2, acetylated histone-H3(ac-H3), p21, cyclin dependent kinase 4(CDK4), and cysteinyl aspartate-specific protease (Caspase)-3. Results Compared to negative control group, the respective inhibition rate of tumor weight in high-dose group, mediate-dose group, low-dose group, and 5-fluorouracil group was 15. 36%(P> 0. 05),40. 27%(P 0. 05)and 55. 63%(P 0. 05), 40. 47%(P 0. 05) and 55. 74%(P<0. 01)respectively.CS055 could significantly inhibit the expression of HDAC1 and HDAC2,upregulate the level of acetylation of histone H3 and the expression of p21,Caspase-3, while downregulate the expression of CDK4 protein. Conclusion CS055 can inhibit the growth of human colon carcinoma xenograft tumor probably by inhibiting the expression of HDAC1 and HDAC2, upregulating the level of histone H3 acetylation and the expression of p21 protein, downregulating the expression of CDK4 protein, which promotes tumor cell cycle arrest, and upregulating the expression of Caspase-3 protein, which induces LoVo cells apoptosis. Key words: Colorectal cancer; Histone deacetylase inhibitor; Apoptosis; Nude mice

Synergistic effect of puerarin and 5-fluorouracil on hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common types of human malignancy worldwide, which is becoming increasingly resistant to traditional drug treatments. Puerarin combined with 5-fluorouracil (5-FU) may be a useful treatment for liver cancer. The primary aim of the present study was to determine whether combined treatment with 5-FU and puerarin is more effective against the hepatocellular carcinoma (HCC) cell line, SMMC7721, than treatment with 5-FU or puerarin alone. The growth inhibition of SMMC7721 cells by puerarin or 5-FU alone or in combination was determined by the Cell Counting Kit-8 assay, in vitro. Apoptotic morphological features and the percentage of apoptotic cells were detected using Hoechst 33258 staining and an Annexin V/PI apoptosis kit, respectively. In addition, a tumor xenograft model was established in nude mice using SMMC7721 cells. Puerarin and 5-FU alone or in combination were injected into the mice, and the inhibition of tumor growth was evaluated by monitoring tumor volume and weight. Treatment with 6,400 or 640 μM 5-FU resulted in growth inhibition of 95.56±0.81 and 75.91±3.54%, respectively. The combination index values were <1 when the fraction of affected cells was between 0.2555 and 0.7420. Furthermore, the percentage of apoptotic cells was markedly increased in the combined treatment group when compared with that of the individual treatment groups, in vitro and in vivo. Individual treatment with puerarin resulted in a tumor volume inhibition rate (IR) of 70.58% and a tumor weight IR of 46.20%. Treatment with 5-FU was found to decrease the tumor volume by 76.26% and tumor weight by 49.86%. In the combined treatment group, the tumor volume and weight IRs were 93.11 and 75.21%, respectively. A marked increase in the inhibition of tumor growth and the number of apoptotic cells in response to combined treatment with puerarin and 5-FU was identified with no observed liver or renal toxicity. These results suggest that puerarin and 5-FU exhibit a synergistic treatment effect on the HCC SMMC7721 cell line.

Yiqi formula enhances the antitumor effects of erlotinib for treatment of triple-negative breast cancer xenografts.

Yiqi formula (YF), a traditional herbal prescription, has long been used to treat triple-negative breast cancer (TNBC) patients. The present study aims to investigate the effects and the related mechanism of YF for treatment of TNBC xenografts. MDA-MB-231 (human TNBC) cells were subcutaneously injected into the second mammary fat pad of 40 female nude mice, which were divided into four groups: control, erlotinib (an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor), YF, and combination (YF plus erlotinib). All treatments were administered orally for 30 days. Inhibition rate of tumor weight by erlotinib, YF, and the combination was 26.47%, 17.24%, and 39.15%, respectively. Western blotting showed that YF, erlotinib, and the combination downregulated p-EGFR (P < 0.01) and p-Akt1 (pT308) (P < 0.05) and upregulated PTEN compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). Similar results were detected by immunohistochemistry. Real-time quantitative PCR showed that YF, erlotinib, and the combination increased PTEN mRNA (P < 0.05, P < 0.01) compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). In conclusion, YF can regulate the main components of the PI3K/Akt pathway in TNBC xenografts. When YF was used in combination with erlotinib, it enhanced the antitumor effects of erlotinib on TNBC xenografts. These findings suggest that YF is suitable to use for the treatment of TNBC patients.

Effect of bone-marrow-derived mesenchymal stem cells on high-potential hepatocellular carcinoma in mouse models: an intervention study

BackgroundThere are two completely contradictory views regarding the impact of human bone-marrow-derived mesenchymal stem cells (hMSCs) on hepatocellular carcinomas (HCCs). The aim of this study was to investigate the effect of hMSC engraftment on HCC tissues in nude mouse models, and assess the effect on metastatic potential of HCC.MethodshMSCs were engrafted into the nude mouse models of high metastatic HCC via the tail vein. The mice in the experimental group were engrafted with hMSCs (5 × 105 cells per mouse) via the tail vein 15 days after inoculation of tumor cells, twice a week, while the animals in the control group were injected with hMSC culture medium (0.2 mL per mouse) via the tail vein. The subcutaneous tumor size was measured using an electronic digital caliper once every 4 days after hMSC engraftment. After 2, 3, 4, 5 and 6 weeks of tumor cell inoculation, the mice were killed and the tumors were collected in their entirety. The tumor weights and body weights of mice were measured, and the tumor inhibition rate was calculated. Quantitative real-time polymerase chain reaction (RT-PCR) was used to determine the expression of metastasis-related genes including osteopontin (OPN), bone sialoprotein (BSP) and integrin α5 subunit (α-V) in the mouse models of high-metastatic HCC, and the expression of apoptosis-related genes including B cell lymphoma/leukemia-2 (Bcl2), Bcl-2 associated X protein (Bax) and caspase 3 in tumor samples.ResultsThe tumor weight inhibition rate was 26.62% at 2 weeks, 52.00% at 3 weeks, 38.20% at 4 weeks, 31.98% at 5 weeks, and 30.23% at 6 weeks. Tumor tissue weight comparison results were significantly lower in the hMSC engraftment groups than in the control group at the second and third weeks. The expression of metastasis-related factors OPN, BSP and α-V gene was downregulated with time. The expression of antiapoptotic gene Bcl2 exhibited an obvious declining tendency, while the expression of apoptotic genes Bax and caspase 3 showed an obvious rising tendency. The expression of α-V and BSP significantly correlated positively with the expression of Bcl2, and negatively correlated with the expression of Bax and caspase 3. The tumor inhibition rate was not significantly correlated with the expression of antiapoptotic and apoptotic factors, and α-V and BSP factors, though it exhibited a significantly negative correlation with the expression of OPN.ConclusionsThe highest tumor inhibition rate was observed 3 weeks after hMSCs engraftment, and the tumor inhibition rate gradually reduced with the progression of time. The metastatic potential of tumor cells was downregulated after hMSC engraftment and hMSCs induce further tumor cells apoptosis. The decrease in the proliferation ability of tumor cells may induce a decline in metastatic potential in tumor cells.

Arsenic trioxide restores ERα expression in ERα-negative human breast cancer cells and its treatment efficacy in combination with tamoxifen in xenografts in nude mice

To study the demethylation effect of arsenic trioxide (As2O3) on ERα-negative human breast cancer MDA-MB-435s cells and its possible mechanisms, and to observe its treatment efficacy in combination with tamoxifen (TAM) after ERα re-expression. MTT assay was used to examine the inhibitory effect of As2O3 treatment alone or in combination with TAM on cell proliferation. A nude mouse xenograft model was used to further examine the treatment efficacy in vivo. MSP was used to detect the methylation status of ERα gene after treated with As2O3 in MDA-MB-435s cells and the transplanted tumor tissues. RT-PCR was used to detect the mRNA expression of DNMT1 and Erα. Western bolt was used to detect the DNMT1 and ERα protein expression. The diameter of xenograft tumors was measured weekly, and the tumor growth curve was drawn. The level of proliferation of the MDA-MB-435s cells was significantly suppressed after treatment with different concentration of As2O3 alone or As2O3 combined with TAM, and the 4 µmol/L As2O3 + TAM treatment for 72 h showed the highest inhibition rate (62.6%). 1, 2, 4 µmol/L As2O3 had demethylation effect on MDA-MB-435s cells, and the DNMT1 mRNA and protein expression was inhibited and accompanied by ERα mRNA and protein re-expression. The unmethylation specific bands of ERα gene were enhanced after treated by As2O3 alone or As2O3 combined with TAM in the xenograft tumors. The expression of DNMT1 mRNA and protein was inhibited, and accompanied by ERα mRNA and protein re-expression. An significant decrease of volume and weight of the xenograft tumors in the As2O3 treated alone or combined with TAM groups was observed compared with those of the normal saline group or TAM alone group (P < 0.05), and the 4 mg/kg As2O3 + TAM group had the highest inhibition rate of tumor weight (79.5%) and volume (76.4%). ERα can be re-expressed in ERα-negative breast cancer MDA-MB-435s cells after treated with As2O3 by inhibiting the DNMT1 activity. MDA-MB-435s cells are re-sensitized to endocrine therapy after ERα re-expression. As2O3 combined with TAM may provide a new therapeutic approach for patients with ERα-negative breast cancer in the clinic.

Pseudolaric acid B inhibits inducible cyclooxygenase-2 expression via downregulation of the NF-κB pathway in HT-29 cells

Pseudolaric acid B (PAB) is a diterpene acid isolated from the root and trunk bark of Pseudolaric kaempferi Gordon. Previous work has found that PAB has anti-inflammatory and anti-tumor effects in xenograft models of human hepatocellular carcinoma. The aim of this study is to evaluate the correlation between anti-cancer and anti-inflammatory effects of PAB and its molecular mechanisms on HT-29 cells. Production of prostaglandin E2 (PGE2) in HT-29 cells was evaluated by ELISA. mRNA of cyclooxygenase-2 (COX-2) was analyzed by RT-PCR assay. High-content screening (HCS) method was adopted to detect the cytokine mixture (CM)-induced transcription activity of NF-κB and STAT3. Western blotting was used to evaluate the protein expression levels of inflammatory mediators induced by CM. After treatment with PAB in various concentrations, the inhibition rate of cell proliferation was measured with sulforhodamine B assays. For the in vivo studies, tumor-bearing models xenografted with HT-29 cells were developed in nude mice, and following oral administration with PAB, tumor inhibition rate was calculated. PAB inhibited the PGE2 production in HT-29 cells significantly (P < 0.05) with similar results detected at the COX-2 mRNA level. Furthermore, PAB suppressed the COX-2 protein expression and significant nuclear translocation of NF-κB and STAT3 induced by CM, which correlated with a concomitant degradation of I-κB and a decrease in constitutive STAT3 phosphorylation (P < 0.05). Moreover, various concentrations of PAB inhibited the proliferation of HT-29 cells in a dose- and time-dependent manner. In vivo, after treatment with PAB for 17 days, the tumor weight of the 50 and 100 mg/kg treated groups was 0.62 ± 0.15 and 0.54 ± 0.06 g, respectively. When compared to the control group (0.82 ± 0.16 g), the inhibition rate of tumor weight was 24.2% at 50 mg/kg (P < 0.05) and 34.7% at 100 mg/kg (P < 0.001). PAB shows potential anti-cancer activity in HT-29 cells, and its molecular mechanisms are related to the anti-inflammatory action.

Anticancer effects of 5-fluorouracil combined with warming and relieving cold phlegm formula on human breast cancer

To investigate the anticancer effects of warming and relieving cold phlegm formula (, WRCP), a Chinese medical mixture composed of the aqueous extracts of Aconitum carmichaeli, Rhizoma bolbostemmatis, Phytolacca acinosa, Panax notoginseng, and Gekko swinhonis Gūenther, combined with 5-fluorouracil (5-FU) on human breast cancer in vivo. Seventy-two Nu/Nu mice inoculated with MDA-MB-231 breast cancer cells were randomized into the control group, 5-FU group, high-dose WRCP (hWRCP) group, medium-dose WRCP (mWRCP) group, low-dose WRCP (lWRCP) group, or combination of mWRCP and 5-FU group in a 1:1:1:1:1:1 ratio. Drug administration was commenced on the day following tumor implantation. The control group was injected daily with normal saline (N.S.) intraperitoneally; the 5-FU group was injected with 5-FU at 30 mg/kg intraperitoneally every third day for a total of 7 treatments; the hWRCP group, mWRCP group and lWRCP group received daily doses of 5, 1, and 0.2 g/kg of WRCP, respectively, by gastric perfusion; and the combination group was treated with 5-FU plus mWRCP on the same schedules as above. All treatments lasted for 22 days. Tumor volume, tumor weight, inhibition rate of tumor weight, necrosis rate of tumor, organ index, and change in body weight of nude mice were measured. The combination group and the hWRCP group had significantly smaller tumor volumes (580±339 mm(3) and 587±249 mm(3) versus 1055±234 mm(3), respectively), lower tumor weights (0.42±0.29 g and 0.52±0.29 g versus 0.80±0.15 g, respectively), and higher tumor necrosis rates (22.7% and 25.6% versus 9.4%, respectively) as compared with the control group (all <0.05). Similar changes were found in the 5-FU, mWRCP, and lWRCP groups when compared with the control group but were not statistically significant, except for the tumor weight for the 5-FU group. The combination group and the hWRCP group had significantly smaller tumor volumes compared with the 5-FU group (778±202 mm(3), both <0.05). The combination group had the highest tumor inhibition rate (47.7%), followed by the hWRCP group (35.2%) and 5-FU group (28.3%). The 5-FU group had a lower body weight increase (1.37±2.06 g versus 5.60±0.72 g, <0.05) and a lower spleen index (4.064±1.774 mg/10 g versus 5.294±1.796 mg/10 g) as compared with the control group, whereas the combination group reversed the changes in the 5-FU group with the body weight increase of 3.52±1.80 g (P <0.05) and spleen index of 7.036±1.599 mg/10 g (P <0.05). The spleen indices in the hWRCP, mWRCP, and IWRCP group were all significantly higher than that in the 5-FU group (P <0.01 or P<0.05). No significant differences in body weight change were observed in WRCP groups compared with the control group P>0.05). The treatment combination of WRCP and 5-FU was more effective in the inhibition of tumor growth than either agent alone and may have potentially additional benefit in improving the general condition and immunity of the mice with human breast cancer cell implants.

Effects of human Livin gene down-regulation by RNA interference on xenograft of transplantation tumor induced by SPC-A1 in nude mice

To explore the in vivo inhibitory effect of Livin gene silencing by RNA interference on xenograft of lung adenocarcinoma SPC-A-1 cells in BALB/C nude mice. Three different BALB/C nude mice models were established by subcutaneously inoculating differently treated SPC-A-1 cells into 3 nude mice groups: the blank control group was inoculated with blank SPC-A-1 cells, while the negative group was inoculated with cells transfected with lentivirus-delivered negative shRNA, the experimental group was inoculated with cells with lentivirus-delivered Livin shRNA. Then the growth of tumors was observed, and the volume and weight of the tumors were measured at different time points. The curve of tumor growth was then described, and the inhibition rate was calculated. Livin gene expression in the tumor tissues was determined by RT-PCR and Immunohistochemistry. Cell apoptosis of tumor tissues was detected by TUNEL. Slower tumor growth, smaller tumor volume and lighter tumor weight were observed in the experimental group as compared to the blank and negative groups (F = 70.509, P < 0.01; F = 12.821, P < 0.01). The inhibition rate of tumor volume was (59.5 ± 3.4)%, and the inhibition rate of tumor weight was (71.1 ± 5.6)%. Livinα mRNA and Livinβ mRNA expressions in the experimental group were significantly lower than the 2 control groups [(37.2 ± 1.6)% versus (63.3 ± 3.8)%, (66.1 ± 2.6)%; (29.4 ± 1.1)% versus (53.2 ± 3.4)%, (52.3 ± 3.1)% (F(α) = 45.309, P < 0.01; F(β) = 30.076, P < 0.01)]. Livin protein expression level was also significantly lower than the blank and the negative groups [(15.3 ± 2.8)% versus (51.3 ± 2.1)%, (52.5 ± 2.5)%, F = 78.92, P < 0.01]. The apoptosis rate in the experimental group was significantly higher than that in the 2 control groups [(35.4 ± 3.2)% versus (5.4 ± 1.3)%, (8.6 ± 1.5)%, F = 14.509, P < 0.01]. The lentivirus-delivered Livin shRNA was shown to inhibit the proliferation of transplantation tumor of lung carcinoma effectively, and Livin may be a target for gene therapy in lung cancer.

Effect of Runing II on the Growth and Metastasis of Transplanted Tumor in Mammary Cancer-bearing Mice and Its Mechanism

To study the effect of Runing II (a Chinese herbal preparation for mammary cancer) on the growth and metastasis of transplanted tumor of mammary cancer MA-891-bearing TA2 mice and its mechanism. The model of mammary cancer MA-891 cell strain transplanted tumor of TA2 mice with lung metastasis were developed to observe the effect of Runing II on the growth and metastasis of the transplanted tumor. The immunohistochemical method and image analysis were adopted to detect the levels of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), and micro-vessel count (MVC) and micro-vessel area (MVA). In the Runing II group, the tumor weight inhibition rate and the lung metastasis inhibition rate were 37.3% and 65.4% respectively, the tumor growth and lung metastasis were obviously inhibited; And the levels of VEGF and VEGFR, MVC and MVA were significantly decreased as compared with those in the tumor-bearing control group (P<0.05). The Chinese herbal preparation Running II can inhibit the metastasis of tumor through inhibiting the angiogenesis, and the mechanism is possibly related with down-regulation of VEGF and VEGFR expression.

Antitumor effect of BRM

Objective: To evaluate the antitumor effect of BRM, a Chinese traditional medical compound herb, and its mechanism. Methods: Tumor inhibition was tested in vivo on mice with subcutaneous transplanted tumor cells in vitro with and MTT method and tumor cell apoptosis test including cell electric morphology, agarose gel electrophoresis and flow cytometry. Results: It was found that BRM could the resistance to the attack of 5×106 of mouse pancreascarcinoma (MPC-83), Ehrlich ascite tumor (EAC), mouse sarcoma (S-180), and mouse glioblastoma (G422) cells, with the inhibition rate of tumor weight of 66.2%, 50.9%, 47.3%, and 38.7% respectively. The inhibition rate of Suzhou human glioma (SHG-44), human breast carcinoma (MCF-7), and human pancreas carcinoma (PANC1) in vitro were 92.2% (BRM 1:10), 85.1% (BRM 1:10), 48.3% (BRM 1:10), respectively. Meanwhile, BRM was able to induce apoptosis in several human tumor cell lines such as SHG-44 and MCF-7 cells. Morphological changes including cell shrinkage and condensation of chromosomes were observed with electric microscope. Agarose gel electrophoresis of DNA from SHG-44 and MCF-7 cells treated with BRM extracts (1:80 to 1:20) for 24 h, 40 h revealed typical DNA “Ladder” pattern. The early stage and middle-late stage apoptosis increased in SHG-44 and MCF-7 cells treated with BRM extracts (BRM 1:160 to 1:40) from 14 h to 48 h by Annexin-V/PI flow cytometry one analysis. Conclusion: BRM has obvious antitumor bioactivity in vivo to MPC-83, EAC, S-180, and G422 mouse cell strains, and in vitro to SHG-44, MCF-7 human cell lines. Water extracts of BRM could induce apoptosis.