siRNA阻断NF-κB信号通路抑制胃癌SGC-7901细胞的增殖及侵袭

Inhibitory effects of siRNA on proliferation and invasiveness of gastric cancer SGC-7901 cells by blocking NF-κB signaling pathway

目的采用RNA干扰(RNA interference,RNAi)技术下调胃癌细胞株SGC-7901中NF-κB p65基因的表达,探讨其对肿瘤细胞增殖活性和侵袭能力的影响。方法利用阳离子脂质体LipofectamineTM2000将化学合成的人NF-κB p65的小干扰RNA(small interference RNA,siRNA)转染入胃癌细胞株SGC-7901中。采用RT-PCR法测定细胞内NF-κB p65mRNA的表达;酶联免疫吸附测定法(ELISA)检测NF-κB亚单位p65的DNA结合活性的改变;采用MTT法测定细胞增殖活性的变化;利用Transwell侵袭实验检测细胞体外侵袭能力的变化。结果与对照组比较,化学合成的人NF-κB p65siRNA能有效地抑制SGC-7901细胞中NF-κB p65mRNA的表达(P<0.05);RelAsiRNA组的p65亚单位与DNA结合活性明显低于对照组(P<0.05),并且RelA siRNA组中SGC-7901细胞的体外侵袭力减弱,增殖活性降低。结论特异的siRNA可以有效阻断NF-κB信号通路,影响人胃癌细胞的增殖活性和体外侵袭能力。

Objective To investigate the inhibitory effects of NF-κB p65 gene targeted small interference RNA(siRNA) on the proliferation and invasiveness of gastric cancer cell line SGC-7901.Methods Chemically synthesized small interference RNA(siRNA) directed against human NF-κB p65 was transfected into gastric carcinoma cell SGC-7901 using cationic liposome LipofectamineTM 2000 as the transfecting agent.The expression of NF-κB p65 gene was detected by RT-PCR.The DNA binding activity of NF-κB subunit p65 was detected by ELISA.Transwell invasiveness test was used to examine the changes of invasive ability of SGC-7901.The proliferation capability was determined by MTT assay.Results RT-PCR results indicated that chemically synthesized siRNA directed against human NF-κB p65 could knock down the transcription and expression of NF-κB p65 gene.The difference between the RelA siRNA group and control group was significant(P<0.05).After transfection,the DNA binding activity of NF-κB p65 in RelA siRNA group was much lower than that of the control group(P<0.05).The invasive and proliferative abilities of SGC-7901 decreased substantially.Conclusion NF-κB p65 siRNA effectively down-regulates the expression of NF-κB p65 gene and decreases the proliferation and invasiveness of SGC-7901cells.