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目的:探讨磷脂酰肌醇-3激酶催化亚基δ(phosphoinositide-3 kinase,catalytic subunit delta,PIK 3CD)基因沉默对胃癌HGC-27细胞体外增殖和迁移的影响及其可能的作用机制。方法:首先采用实时荧光定量PCR法检测人胃癌HGC-27细胞中PIK 3CD基因的表达水平;然后将MSCV-PIK3CD-sh RNA重组质粒转染到HGC-27细胞中,采用实时荧光定量PCR和蛋白质印迹法鉴定PIK 3CD基因沉默效果;采用CCK-8试剂盒和Transwell小室法分别检测PIK 3CD基因沉默对胃癌HGC-27细胞增殖和迁移的影响;蛋白质印迹法检测PIK3CD基因沉默后其下游相关的Akt信号通路分子的表达情况。结果:PIK3CD在胃癌HGC-27细胞中高表达。MSCV-PIK3CD-sh RNA转染入PIK3CD基因高表达的胃癌HGC-27细胞后,PIK3CD表达被明显下调(P<0.05),细胞增殖和细胞迁移能力明显降低(P<0.05)。PIK 3CD基因沉默对Akt表达无明显影响,但可明显抑制p-Akt的表达水平(P<0.05)。结论:PIK3CD可能通过激活Akt信号转导通路,促进胃癌HGC-27细胞的体外增殖和迁移。
OBJECTIVE: To investigate the effect of phosphoinositide 3 kinase (PI3K) gene silencing on proliferation and migration of gastric cancer cell line HGC-27 in vitro and its possible mechanism. Methods: The expression level of PIK 3CD gene in human gastric cancer HGC-27 cells was detected by real-time fluorescence quantitative PCR. The recombinant plasmid of MSCV-PIK3 CD-sh RNA was transfected into HGC-27 cells. Real-time PCR and protein The effect of PIK 3CD gene silencing on the proliferation and migration of gastric cancer cell line HGC-27 was detected by CCK-8 kit and Transwell chamber assay. Western blotting was used to detect the downstream Akt Signaling molecule expression. Results: PIK3CD was highly expressed in gastric cancer HGC-27 cells. After transfection of MSCV-PIK3CD-sh RNA into HGC-27 cells with high expression of PIK3CD gene, PIK3CD expression was significantly downregulated (P <0.05), and cell proliferation and cell migration were significantly decreased (P <0.05). PIK 3CD gene silencing had no significant effect on Akt expression, but significantly inhibited the expression of p-Akt (P <0.05). Conclusion: PIK3CD may promote the proliferation and migration of gastric cancer HGC-27 cells through activation of Akt signal transduction pathway.