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目的构建针对信号转导子和转录激活子3(signal transducer and activator of transcription 3,STAT3)的小干扰RNA(siRNA)表达载体STAT3-siRNA,研究siRNA干扰STAT3基因表达后对人胃癌细胞株HGC-27细胞增殖作用的影响。方法设计并合成两个STAT3-siRNA片段,经脂质体介导转染HGC-27细胞,RT-PCR和Westernblot检测转染后HGC-27细胞中STAT3 mRNA和蛋白的表达,MTT法检测STAT3-siRNA对HGC-27细胞增殖作用的影响。结果酶切及测序鉴定证实STAT3-siRNA1和STAT3-siRNA 2表达载体均构建成功。RT-PCR和Western blot结果显示,转染后STAT3 mRNA和蛋白的表达量均下降,与空白对照组及si-NC转染对照组相比,差异具有统计学意义(P<0.01),其中以STAT3-siRNA1干扰效果更明显。MTT结果显示,转染STAT3-siRNA1后,HGC-27细胞的增殖能力明显下降,与si-NC转染对照组相比,差异具有统计学意义(P<0.01)。结论本研究构建的STAT3-siRNA表达载体可有效抑制STAT3在胃癌细胞株HGC-27中的表达,并抑制胃癌细胞的生长。
Objective To construct a small interfering RNA (siRNA) expression vector STAT3-siRNA targeting signal transducers and activator of transcription 3 (STAT3) to study the effect of siRNA on STAT3 gene expression in human gastric cancer cell line HGC- 27 cells proliferation effect. Methods Two STAT3-siRNA fragments were designed and synthesized. HGC-27 cells were transfected by liposome. STAT3 mRNA and protein expression in HGC-27 cells were detected by RT-PCR and Western blotting. STAT3- Effect of siRNA on proliferation of HGC-27 cells. Results Enzyme digestion and sequencing confirmed that STAT3-siRNA1 and STAT3-siRNA 2 expression vectors were constructed successfully. The results of RT-PCR and Western blot showed that the expression of STAT3 mRNA and protein decreased after transfection compared with the blank control group and si-NC transfected control group, the difference was statistically significant (P <0.01) STAT3-siRNA1 interference effect is more obvious. The results of MTT showed that the proliferation of HGC-27 cells was significantly decreased after transfected with STAT3-siRNA1, which was significantly different from that of the control group transfected with si-NC (P <0.01). Conclusion The STAT3-siRNA expression vector constructed in this study can effectively inhibit STAT3 expression in gastric cancer cell line HGC-27 and inhibit the growth of gastric cancer cells.