摘要
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 analysis the expression of VEGF - C,NF - KB,bcl - 2 gene by inhibiting VEGF - C expression of tumor cells with RNA interference （RNAi） technique. Methods According to the encoding sequence mRNA of human VEGF - C,the target site for RNAi was designed, siRNA was transfected by liposome and the expression of VEGF - C, NF - KB, bcl - 2 mRNA was further observed by RT - PCR. Results The expression of VEGF - C of HeLa cell lines treated with siRNA for 24h,48h was significantly inhibited,with in24hSO. 63% ±0.24% （P〈O.O01） and in48h38.9% ＋0.85%（P〈0. O1）.The expression of NF-KB was also down-regulated in 24h,48h,with in 24h 37.55% ＋ 2.76% （P 〈0.05） and in 48h 30.5% ±3.82% （P =0. 056）. Thus the expression of bcl - 2 was also inhibited in 24h,48h,with in 24h 76. 95%±1 .91% （P〈0.01） and in 48h 64. 11% ＋2.96% （P〈0.05）.Conclusion After being treated with VEGF - C siRNA of the HeLa cells, the expression of VEGF - C is effectively inhibited, NF - KB may be down - regulated and anti - apoptosis bcl - 2 gene is effectively inhibited.
出处
《医学研究杂志》
2011年第11期58-62,共5页
Journal of Medical Research
基金
浙江省卫生厅科技基金资助项目(2008196B)
浙江省台州科技基金课题(08YK14)
