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目的:探讨扶正抗癌方抑制PC9细胞转移的分子机制。方法:台盼蓝活力检测探索扶正抗癌方对PC9细胞增殖的影响;transwell实验探索扶正抗癌方对PC9细胞侵袭和迁移的影响;基质金属蛋白酶-9(MMP-9)活力检测探索扶正抗癌方对MMP-9活力的影响,Real-time quantitative PCR探索扶正抗癌方对MMP-9 mRNA表达的影响,蛋白质印迹法探索扶正抗癌方对MMP-9、磷酸化的信号转导子及转录激活子3[p-STAT3(Tyr705)]蛋白表达的影响并探讨两者的关系。结果:扶正抗癌方以浓度依赖性抑制PC9细胞的增殖、侵袭和迁移(P<0.05);以浓度依赖性下调MMP-9活力、mRNA和蛋白的表达(P<0.05),以时间依赖性下调p-STAT3(Tyr705)蛋白的表达(P<0.05);过表达STAT3逆转了扶正抗癌方对MMP-9的下调(P<0.05)。结论:扶正抗癌方通过抑制STAT3的表达下调MMP-9,进而抑制PC9细胞的转移。
Objective: To investigate the molecular mechanism of Fuzheng Kangai Fang inhibiting PC9 cell metastasis. Methods: Trypan blue assay was used to explore the effect of Fuzheng anticancer agent on proliferation of PC9 cells. Transwell experiment was conducted to explore the effect of Fuzheng Kangai Fang on PC9 cell invasion and migration. Matrix metalloproteinase-9 (MMP-9) The influence of cancer on the activity of MMP-9, Real-time quantitative PCR to explore the effect of Fuzheng anticancer on the expression of MMP-9 mRNA, Western blotting explore Fuzheng anti-cancer on MMP-9, phosphorylation of signal transducers and Transcriptional activator 3 [p-STAT3 (Tyr705)] protein expression and explore the relationship between the two. Results: Fuzheng anticancer drug inhibited the proliferation, invasion and migration of PC9 cells in a concentration-dependent manner (P <0.05), and down-regulated the expression of MMP-9, mRNA and protein in a concentration- Down-regulated the expression of p-STAT3 (Tyr705) protein (P <0.05); Overexpression of STAT3 reversed the down-regulation of MMP-9 by Fuzheng Kangai Fang (P <0.05). Conclusion: Fuzheng Kangai Fang down-regulates the expression of STAT3 and down-regulates the expression of MMP-9, thereby inhibiting the metastasis of PC9 cells.