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目的观察shRNA下调胰腺癌Panc-1细胞STAT3 mRNA表达后对吉西他滨化疗敏感性的影响。方法构建靶向STAT3 mRNA的shRAN和阴性质粒,分别转染Panc-1细胞(A组和B组);另将非转染的正常Panc-1细胞分为C组和D组。A,B,C三组细胞与吉西他滨共孵育。RT-PCR和Western blot检测Panc-1中STAT3 mRNA和蛋白的表达;MTT法检测Panc-1细胞增殖能力;流式细胞术检测细胞周期。结果 A组STAT3 mRNA水平下降了61.9%,STAT3蛋白水平下降了85.7%(P<0.01),细胞增殖能力低于B、C组(P<0.01)。与B、C组比较,A组G1期细胞比率增加(P<0.05),细胞存活率降低(P<0.01)。结论靶向STAT3 mRAN的shRNA可特异性降低Panc-1细胞中STAT3 mRNA及其蛋白的表达,抑制Panc-1细胞增殖,使其阻滞于G1期,从而增强吉西他滨对G1期细胞的细胞毒效应,增加吉西他滨的化疗敏感性。
Objective To investigate the effect of shRNA on the sensitivity of gemcitabine to STAT3 mRNA expression in Panc-1 pancreatic cancer cells. Methods The shRAN and negative plasmids targeting STAT3 mRNA were constructed and transfected into Panc-1 cells (group A and group B) respectively. The non-transfected normal Panc-1 cells were divided into group C and group D. A, B, C three groups of cells were incubated with gemcitabine. The expression of STAT3 mRNA and protein in Panc-1 cells was detected by RT-PCR and Western blot. The proliferation of Panc-1 cells was detected by MTT assay. The cell cycle was detected by flow cytometry. Results STAT3 mRNA level decreased by 61.9% and STAT3 protein level decreased by 85.7% (P <0.01) in group A, but lower than that in groups B and C (P <0.01). Compared with group B and group C, the percentage of cells in G1 phase in group A increased (P <0.05), and the cell survival rate decreased (P <0.01). Conclusion shRNA targeting STAT3 mRAN can specifically down-regulate the expression of STAT3 mRNA and protein in Panc-1 cells, inhibit the proliferation of Panc-1 cells and arrest the G1 phase cells, thereby enhancing the cytotoxic effect of gemcitabine on G1 phase cells , Increasing the chemosensitivity of gemcitabine.