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目的研究由2型重组腺相关病毒(rAAV2)介导的靶向缺氧诱导因子2(EPAS1/HIF-2)的小干扰RNA(siR-NA)对人胰腺癌PANC-1细胞凋亡和增殖的影响。方法将人胰腺癌细胞株PANC-1分为实验组、阴性对照组(阴性组)和空白对照组(空白组),实验组和阴性组分别转染rAAV2-EPAS1-siRNA和rAAV2-EGFP病毒,空白组不转染病毒,进行常氧和缺氧培养。RT-PCR和Western blot检测EPAS1基因表达,Annexin V-FITC/PI双染检测细胞凋亡,MTT法检测细胞增殖能力。结果在常氧和缺氧状态下,实验组细胞EPAS1 mRNA和蛋白表达受抑制,显著低于阴性组和空白组,差异均有高度统计学意义(P<0.001)。在缺氧状态下,实验组细胞的凋亡率明显升高,增殖受抑制,与阴性组和空白组比较,差异有统计学意义(P<0.05)。结论通过siRNA抑制缺氧状态下胰腺癌细胞中EPAS1基因表达,能诱导胰腺癌细胞凋亡,抑制胰腺癌细胞增殖。
Objective To investigate the effects of rAAV2-mediated siRNA targeting siR-NA targeting hypoxia-inducible factor 2 (HAS-2) on apoptosis and proliferation of human pancreatic cancer PANC-1 cells Impact. Methods Human pancreatic cancer cell line PANC-1 was divided into experimental group, negative control group (negative group) and blank control group (blank group). The experimental group and the negative group were transfected with rAAV2-EPAS1-siRNA and rAAV2- The blank group did not transfect the virus for normoxia and hypoxia culture. The expression of EPAS1 gene was detected by RT-PCR and Western blot. Apoptosis was detected by Annexin V-FITC / PI double staining. Cell proliferation was measured by MTT assay. Results Under normoxia and hypoxia, the expression of EPAS1 mRNA and protein in the experimental group was significantly lower than that in the negative group and the blank group (P <0.001). Under hypoxia, the apoptotic rate of the experimental group was significantly increased, and the proliferation was inhibited. Compared with the negative group and the blank group, the difference was statistically significant (P <0.05). Conclusion siRNA inhibits the expression of EPAS1 in pancreatic cancer cells under hypoxia, which can induce the apoptosis of pancreatic cancer cells and inhibit the proliferation of pancreatic cancer cells.