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目的通过RNA干扰技术抑制宫颈癌细胞VEGF-C表达,探讨干扰后VEGF-C、NF-κB、bcl-2基因的表达。方法根据人VEGF-C mRNA编码序列,设计RNA干扰的靶点,并用脂质体转染人宫颈癌HeLa细胞,通过RT-PCR法观察转染后肿瘤细胞VEGF-C、NF-κB、bcl-2基因的变化。结果转染siRNA 24h、48h可以使HeLa细胞VEGF-C mRNA含量降低,在24h降低80.63%±0.24%(P<0.001);在48h降低38.9%±0.85%(P<0.01);NF-κB mRNA含量也分别降低,在24h降低37.55±2.76%(P<0.05);在48h降低30.5%±3.82%(P=0.056);bcl-2mRNA含量同时分别降低,在24h降低76.95%±1.91%,(P<0.01);在48h降低64.11%±2.96%,(P<0.05))。结论脂质体介导的VEGF-C siRNA转染HeLa细胞后,可以有效抑制VEGF-C的表达;可能通过下调转录因子NF-κB,抑制抗凋亡基因bcl-2的表达。
Objective To investigate the expression of VEGF-C, NF-κB and bcl-2 gene in cervical cancer cells by RNAi technique. Methods According to the coding sequence of human VEGF-C mRNA, the target of RNA interference was designed and transfected into HeLa cells by lipofectamine. The expression of VEGF-C, NF-κB and bcl- 2 gene changes. Results The expression of VEGF-C mRNA in HeLa cells decreased by 80.63% ± 0.24% (P <0.001) at 48 h and decreased by 38.9% ± 0.85% at 48 h (P <0.01) (P <0.05), decreased by 30.5% ± 3.82% at 48h (P = 0.056), while the content of bcl-2 mRNA decreased at the same time, decreased by 76.95% ± 1.91% at 24h P <0.01), decreased by 64.11% ± 2.96% at 48h (P <0.05). Conclusion VEGF-C siRNA can effectively inhibit the expression of VEGF-C after transfection with VEGF-C siRNA. It may inhibit the expression of anti-apoptotic gene bcl-2 by down-regulating the transcription factor NF-κB.