RNAi抑制ALK1对人胃腺癌SGC-7901细胞增殖的影响

摘要

Background and purpose: ALK1 (activin receptor-like kinase-1) can promote the proliferation of endothelium cell, and stimulate angiogenesis in tumors.This study aimed to investigate the effect of small interfering RNA (siRNA) on the inhibiting of ALK1 and the proliferation of gastric cancer cell line SGC-7901 after ALK1 silencing.Methods: Three small interference RNA that targeting ALK1 gene were designed and cloned into the pGenesil-1 plasmid.Then the pGenesil-l-ALKl-siRNA vector was transfectd into human gastric cancer cell line SGC-7901.The expression of ALK1 mRNA and protein were detected by Real-time PCR and Western blot respectively.The cell proliferation was measured by MTT and flow cytometry method.Results: Comparing to the negative group, the third ALK1 interference RNA could silence ALK1 gene expression in mRNA and protein level significantly.And after ALK1 silencing, the proliferation of SGC-7901 was inhibited.Conclusion: ALK1 takes a pivotal role in the cancer proliferation and may be a promising target in cancer therapy.%背景与目的:人活化素受体样激酶-1(activin receptor-like kinase-1,ALK1)可促进内皮细胞增殖,与肿瘤内血管生成密切相关.本研究采用RNA干扰技术(RNA interference,RNAi)抑制ALK1表达,观察抑制效果以及抑制ALK1表达后对人胃癌细胞增殖的影响.方法:设计3条靶向ALK1基因的小干扰RNA(small interfering RNA,siRNA),克隆到真核表达质粒pGenesil-1,转染人胃癌细胞株SGC-7901.分别采用Real-time PCR和Western blot技术检测胃癌细胞ALK1 mRNA及其蛋白表达的变化,并检测细胞周期及生长曲线.结果:与阴性对照相比,第3条ALK1干扰RNA在mRNA及蛋白水平均可明显下调ALK1基因表达,并抑制SGC-7901细胞增殖.结论:RNAi技术导致的ALK1表达下调并抑制胃癌细胞增殖,ALK1可作为胃癌抗肿瘤治疗的理想靶点.