CAJ | 학술논문

目的应用RNA干扰技术抑制瞬时受体电位通道(TRPC1)基因的表达,观察TRPC1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TRPC1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TRPC1 mRNA和蛋白表达水平.通过MTT、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TRPC1基因的siRNA明显抑制了目的基因的表达.体外实验表明,靶向TRPC1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TRPC1基因的siRNA导致了细胞周期G0/G1期阻滞(转染NC-siRNA 48 h后,A549细胞处于G0/G1期的细胞比例为54.7％±5.8％,而转染TRPC1-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TRPC1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TRPC1基因的表达可显著抑制肺癌细胞的增殖和侵袭能力.
목적 응용RNA간우기술억제순시수체전위통도(TRPC1)기인적표체,관찰TRPC1대폐선암A549세포증식화침습능력적영향.방법 장경화학수식적TRPC1특이성소간우성RNA(siRNA)전염A549세포,채용형광정량PCR화단백질인적법검측세포TRPC1 mRNA화단백표체수평.통과MTT、세포주기검측화침습실험,관찰A549세포악성생물학행위적개변.결과 파향TRPC1기인적siRNA명현억제료목적기인적표체.체외실험표명,파향TRPC1기인적siRNA전염A459세포후,세포증식억제솔체34.7％,이비특이성siRNA미능영향세포증식.세포주기검측결과현시파향TRPC1기인적siRNA도치료세포주기G0/G1기조체(전염NC-siRNA 48 h후,A549세포처우G0/G1기적세포비례위54.7％±5.8％,이전염TRPC1-siRNA적세포처우G0/G1기적세포비례위71.0％±7.0％,P＜0.05).동시,당TRPC1기인표체피억제시,A459세포침습능력명현하강(P＜0.05).결론 억제TRPC1기인적표체가현저억제폐암세포적증식화침습능력.
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 adenocarcinoma 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 realtime 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 cells with an inhibitory rate of 34.7％ while negative control siRNA had no effect on cell proliferation.Flow cytometric analysis showed that siRNA targeting TRPC1 increased the number of cells in G0/G1 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.