针对CD_(44)V_3的短发夹RNA抑制肺癌A549细胞体外增殖

Inhibitory effects of shRNA targeting CD_(44)V_3 on proliferation ability of human lung carcinoma cell A549 in vitro

目的:研究针对CD44V3的短发夹RNA(short hairpin RNA,shRNA)对体外培养的人肺腺癌A549细胞CD44V3表达及增殖能力的影响。方法:设计和构建带有U6启动子的、能产生针对CD44V3的shRNA的RNA干扰表达质粒,转染至A549细胞,以RT-PCR和Western blot检测A549细胞转染前后CD44V3表达,以MTT实验和软琼脂细胞集落形成实验比较A549细胞转染前后增殖能力。结果:和转染前相比,转染了RNA干扰表达质粒的A549细胞CD44V3mRNA和蛋白表达、以及增殖能力皆明显降低。结论:针对CD44V3的短发夹RNA能明显抑制人肺腺癌A549细胞体外增殖能力。

OBJECTIVE：To investigate the effect of short hairpin RNA （shRNA） targeting CD44 V3 on CD44 V3 expression and proliferation ability of human lung carcinoma cell line A549 in vitro. METHODS： RNA interference plasmid that include U6 promoter and can express shRNA of CD44 V3 was designed, constructed, and transfected into A549 cell line. CD44 V3 expression was examined by reverse transcription polymerase chain reaction （RT-PCR） and western blot. Proliferation ability was examined by MTT assay and clone formation assay. RESULTS： After plasmid transfected, CD44 V3 expression in A549 cell and proliferation ability of A549 cell decreased significantly. CONCLUSION： The shRNA targeting CD44 V3 could inhibit efficiently proliferation ability of human lung carcinoma cell A549 in vitro.