关键词: 蛋白激酶D, shRNA干扰, 细胞程序性死亡

Abstract:

 Objective  To explore the effect of silencing protein kinase D (PKD)-2 on Tca8113 cell proliferation, programmed cell death,and chemosensitivity. Methods  The stable celllines of pkd-2 gene silencing and empty vector plasmid group were established. The proliferation and 50% inhibitory concentration (IC50) of shRNA-mediated Tca8113 chemotherapy drugs were detected through methyl thiazolyl tetrazolium assay (MTT). The programmed cell death rate and sensitivity to Tca8113 chemotherapy drugs before or after pkd-2 gene silencing were measured through flow cytometry. P-glycoprotein (P-gp) expression of pkd-2 silencing cells was identified by immunohistochemical methods. Results Stable cell lines of pkd-2 gene silencing were established. Compared with parental cells, the proliferation of shRNA-mediated Tca8113 was not significantly different,but its IC50 was lower. Meanwhile,cell programmed death rate and sensitivity to chemotherapeutic drugs of shRNA-mediated Tca8113 significantly increased. Compared with wild group Tca8113, a significant decrease in P-gp ex-pression was induced by chemotherapy drugs with shRNA-pkd-2 gene silencing. Conclusion  The pkd-2 gene of shRNA interference silencing Tca8113 promotes programmed cell death of Tca8113, reduces the IC50 of the chemotherapy drugs, and significantly improves the sensitivity of Tca8113 to chemotherapeutic drugs while reducing the expression of P-gp.

Key words: protein kinase D, shRNA interference, programmed cell death