siRNA阻断NF-κB信号通路抑制胃癌SGC-7901细胞的增殖及侵袭

摘要

目的 采用RNA干扰(RNAinterference,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LipofectamineTM 2000将化学合成的人NF-κB p65的小干扰RNA(small interference RNA,siRNA)转染人胃癌细胞株SGC- 7901中.采用RT-PCR法测定细胞内NF-κB p65 mRNA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 p65 siRNA能有效地抑制SGC-7901细胞中NF-κB p65 mRNA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.%To investigate the inhibitory effects of NF-kB p65 gene targeted small interference RNA (siRNA) on the proliferation and ivasiveness of gastric cancer cell line SGC-7901. Methods Chemically synthesized small interference RNA (siRNA) directed against human NF-kB p65 was transfected into gastric carcinoma cell SGC-7901 using cationic liposome LipofectamineTM 2000 as the transfecting agent. The expression of NF-kB p65 gene was detected by RT-PCR, The DNA binding activity of NF-kB 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-kB p65 could knock down the transcription and expression of NF-kB 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-kB p65 in RelA siRNA group was much lower than that of the control group (F< 0.05). The invasive and proliferative abilities of SGC-7901 decreased substantially. Conclusion NF-kB p65 siRNA effectively down-regulates the expression of NF-kB p65 gene and decreases the proliferation and invasiveness of SGC-790lcells