Inhibition of kallikrein 10 gene effecting on the proliferation and apoptosis of colon cancer cell and its chemosensitivity to cisplatin

Abstract

Objective To study the effect of KLK10 on the proliferation and apoptosis of colon cancer cell and its chemosensitivity to cisplatin (DDP) by small interfering RNA (siRNA) targeting KLK10. Methods KLK10 specific siRNA was transfected into colon cancer cell lines HRT-S1 and HCT116 by Lipofectamine® RNAi MAX transfection reagent. real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression level of KLK10 mRNA in NCM460 and HRT-S1 and HCT116 colon cancer cell lines. The expression levels of Cyclin D1 and B cell lymphoma/leukemia-2 (bcl-2) protein were determined by Western blotting. Respectively, cell proliferation, apoptosis and half maximal inhibitory concentration (IC50) of cisplatin were detected by methyl thiazol tetrazolium (MTT) and flow cytometry (FCM) in vitro. Results Compared with those negative control groups, KLK10 mRNA level deceased in experimental group in HRT-S1 and HCT116 (P=0.002); Cyclin D1 protein level deceased in HCT116 (P=0.027); bcl-2 protein deceased in HRT-S1 and HCT116 (P=0.038 and 0.016); The IC50 of cisplatin in experimental group was lower than that in the other groups in HRT-S1 and HCT116 (P=0.032 and 0.022), the cells proliferation was suppressed. Apoptosis rate was significantly higher (P=0.028 and 0.036). Conclusion KLK10 gene suppression can significantly inhibit colon cancer cell proliferation ability, can promote tumor cell apoptosis, enhance the sensitivity of tumor cells to DDP, by inhibitting the expression of Cyclin D1 and bcl-2; KLK10 gene is an important target in the treatment of colon cancer. Key words: Colon cancer; Kallikrein 10; Gene