Karimzadeh, L. | Search bioRxiv

bioRxiv2017

Anti-mutagenic and synergistic cytotoxic effect of cisplatin and Honey Bee venom on 4T1 invasive mammary carcinoma cell line

Honey Bee Venom has various biological activities such as inhibitory effect on several types of cancer. Cisplatin is an old and potent drug to treat the most of cancer. Our aims in this study were determination of the anti-mutagenic and cytotoxic effects of HBV on mammary carcinoma, lonely and in combination with cisplatin. In this study 4T1 cell line were cultured and incubated at 37 C in humidified CO2-incubator. The cell viabilities were examined by MTT assay. Also HBV was screened for its anti-mutagenic activity against sodium azide by Ames test. The result showed that 6g/ml HBV, 20g/ml cisplatin and 6g/ml HBV with 10g/ml cisplatin can induce an approximately 50% 4T1 cell death. 7mg/ml HBV with the inhibition of 62.76% sodium azide showed high potential in decreasing the mutagenic agents. MTT assay demonstrated that HBV and cisplatin can cause cell death in a dose-dependent manner. The cytotoxic effect of cisplatin is also promoted by HBV. Ames test results indicated that HBV can inhibit sodium azide as a mutagenic agent. Anti-mutagenic activity of HBV was increased significantly in presence of S9 mix. Hence, our findings reveal that HBV can enhance the cytotoxic effect of cisplatin drug and it has cancer preventing effects.

pharmacology and toxicology↗

bioRxiv2017

shRNA mediated inhibition of Cdc42 gene expression in Calu-6 lung cancer cells

Background Information: RNAi technique as a new strategy in gene therapy is the effective gene silencing method. Cdc42 is a member of Rho GTPases involving in lung cancer cells migration and proliferation. In the present study, targeting and inhibiting the Cdc42 expression in Calu-6 cells was investigated. Recombinant lentiviral particles were produced by co-transfection of pMD2.G, psPAX2 and pGFP-C-shLenti plasmid in 293T packaging cells. Calu-6 cells were transduced by recombinant lentiviruses using polybrene. GFP-fluorescence microscopy and MTT assay were used to assess the Calu-6 target cell transduction and rate of lentiviral transduced cells proliferation, respectively. Real time PCR was performed to compare the expression of Cdc42 gene before and after shRNA delivery. Results: GFP-fluorescence microscopy analysis showed that Calu-6 cells were successfully transduced with recombinant lentiviral expressing shRNA-Cdc42. The viability of transduced cells was reduced within 72, 96 and 120 hours of transduction process. Real time PCR analyze showed the significant reduction of Cdc42 gene expression. Conclusions: Lentiviral vectors may be reasonable tools for this gene delivery due to stable expression of silencing RNA. Inhibition of Cdc42 expression by lentiviral mediated shRNA delivery could be an effective method to inhibit proliferation of lung cancer cells. Significance: Over expression of Cdc42 gene have been seen in lung cancer. This make Cdc42 gene a key target in treatment of cancer. On the other hand, gene therapy is a proper method to modulate gene expression. It seems modulation of cdc42 gene expression by gene therapy accompany with proper vehicles creating hopes in treatment of lung cancer.

genomics↗