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目的探讨应用RNA干扰技术抑制胃癌细胞组织因子(TF)基因表达对胃癌侵袭能力的影响。方法构建pSUPER/TF-siRNA重组载体,并将其转染入胃癌细胞SGC-7901中;RT-PCR和Western blot检测转染前后TF的表达水平;应用Boyden小室检测对胃癌细胞的侵袭能力。结果双酶切鉴定重组载体构建成功。RT-PCR和Western blot检测显示pSUPER/TF-siRNA1重组载体明显抑制SGC-7901细胞TF基因的表达(P<0.05)。Boyden小室体外侵袭实验证实,转染pSUPER/TF-siRNA1质粒的SGC-7901穿膜细胞均数较其他无干扰组的细胞均数明显减少(P<0.05)。结论以TF基因作为靶点应用RNA干扰技术抑制其表达可降低胃癌细胞体外侵袭移能力,故其有望成为胃癌治疗的一个新靶点。
Objective To investigate the effect of RNA interference on the invasion of gastric cancer cells by inhibiting the expression of tissue factor (TF) gene in gastric cancer cells. Methods The pSUPER / TF-siRNA recombinant vector was constructed and transfected into SGC-7901 cells. The expression of TF was detected by RT-PCR and Western blot. The invasive capacity of gastric cancer cells was detected by Boyden chamber. Results Double enzyme digestion of the recombinant vector was successfully constructed. RT-PCR and Western blot showed that pSUPER / TF-siRNA1 recombinant vector significantly inhibited TF gene expression in SGC-7901 cells (P <0.05). Boyden chamber invasion assay confirmed that transfection of pSUPER / TF-siRNA1 plasmid SGC-7901 cells transmembrane cells were significantly reduced compared with other non-interfering cells (P <0.05). Conclusion The use of TF gene as a target to suppress RNAi expression can reduce the invasion and metastasis potential of gastric cancer cells, so it is expected to become a new therapeutic target for gastric cancer.