SiRNA沉默TRPC1基因对肺腺癌A549细胞增殖和侵袭的影响  

Knocking down TRPC1 expression by siRNA inhibits proliferation and invasiveness of human lung adenocarcinoma cell A549 in vitro

作　　者：张骏[1] 朱震[1] 李雄伟[1] 张忠夫[1] 

机构地区：[1]解放军第一一七医院外科,杭州310013

出　　处：《中华医学杂志》2013年第28期2241-2243,共3页National Medical Journal of China

摘　　要：目的应用RNA干扰技术抑制瞬时受体电位通道（TRPCI）基因的表达，观察TRPCI对肺腺癌A549细胞增殖和侵袭能力的影响。方法将经化学修饰的TRPCI特异性小干扰性RNA（siRNA）转染A549细胞，采用荧光定量PCR和蛋白质印迹法检测细胞TRPCImRNA和蛋白表达水平。通过MTr、细胞周期检测和侵袭实验，观察A549细胞恶性生物学行为的改变。结果靶向TRPCI基因的siRNA明显抑制了目的基因的表达。体外实验表明，靶向TRPCI基因的siRNA转染A459细胞后，细胞增殖抑制率达34．7％，而非特异性siRNA未能影响细胞增殖。细胞周期检测结果显示靶向TRPCI基因的siRNA导致了细胞周期G0／G1期阻滞（转染NC—siRNA48h后，A549细胞处于G0／G1期的细胞比例为54．7％±5．8％，而转染TRPCI-siRNA的细胞处于G0／G1期的细胞比例为71．0％±7．0％，P〈0．05）。同时，当TRPCI基因表达被抑制时，A459细胞侵袭能力明显下降（P〈0．05）。结论抑制TRPCI基因的表达可显著抑制肺癌细胞的增殖和侵袭能力。Objective To evaluate the effects of down-regulated expression of transient receptor potential canonical （TRPC1） by RNA interference （RNAi） on proliferation and invasiveness of human lung adenoearcinoma cell A549 in vitro. Methods A549 cells were transfected with chemically synthesized small interfering RNA （siRNA） targeting TRPC1 gene. The mRNA and protein of TRPC1 were analyzed by real- time polymerase chain reaction （PCR） and Western blot respectively. To assess malignant phenotypes of transfected A549 cells, the assays of methyl thiazolyl tetrazolium （MTT） , cell cycle and cell invasion were performed. Results siRNA targeting TRPC1 dramatically suppressed TRPC1 expression. In vitro study showed that siRNA targeting TRPC1 significantly inhibited cell proliferation of A549 ceils with an inhibitory rate of 34. 7% while negative control siRNA had no effect on cell proliferation. Flow eytometric analysis showed that siRNA targeting TRPC1 increased the number of ceils in G0/Gl phase （P 〈 0. 05）. Moreover, a knockdown of TRPC1 expression effectively inhibited cell invasiveness in A549 cells （P 〈 0. 05 ） . Conclusion Knocking down TRPC1 expression can inhibit proliferation and invasiveness of A549 cells in vitro.

关 键 词：肺肿瘤 TRPC1 RNA 小分子干扰 

分 类 号：R734.2[医药卫生—肿瘤]