hFXYD6反义核酸对胆管癌细胞体外增殖和侵袭能力的影响  

Effect of human antisense FXYD6 on the proliferation and invasion of human cholangiocarcinoma cells in vitro

作　　者：窦春青[1] 岳鑫[2] 周宁新[1] 

机构地区：[1]中国人民解放军总医院全军肝胆外科研究所,北京市100853 [2]北京大学神经科学研究所,北京市100083

出　　处：《世界华人消化杂志》2007年第9期929-935,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金;No.30471699

摘　　要：目的:研究hFXYD6基因反义核酸对人胆管癌QBC939细胞体外增殖和侵袭能力的影响.方法:构建hFXYD6基因反义核酸真核表达载体pcDNA3.1(-)/hFXYD6(-)并转染人胆管癌QBC939细胞,同时设pcDNA3.1(-)空载体对照组和空白对照组.SYBR GreenⅠ荧光定量RT- PCR和免疫组化分别检测hFXYD6 mRNA及蛋白表达;MTT、平板克隆形成实验检测细胞体外增殖活性:流式细胞仪检测细胞周期;Transwell侵袭小室模型检测细胞体外侵袭能力.结果:与空白组和空载组比较,反义组细胞hFXYD6 mRNA及蛋白表达量降低;细胞群体倍增时间增加(46.8 h vs 34.5 h,35.3 h),细胞克隆形成率降低(24.3%±5.3% vs 61.0%±8.5%,58.0%±5.6%,P<0.001);细胞周期中G1期细胞比例明显升高(66.4%±2.9% vs 33.5%±2.3%,39.4%±3.7%,P<0.001),S期比例明显减少(18.6%±1.6% vs 36.2%±2.1%,34.1%±1.6%,P<0.001);Transwell侵袭小室中24 h穿膜细胞数无显著改变.空白组和空载组细胞间均无明显差异.结论:hFXYD6反义核酸抑制人胆管癌QBC939细胞体外增殖能力,但对其侵袭能力无明显作用.AIM： To investigate the effect of human antisense FXYD6 （FXYD domain containing ion transport regulator 6） on the proliferation and invasion of human cholangiocarcinoma cell line QBC939 in vitro. METHODS： Human cholangiocarcinoma cell line QBC939 was transfected with the plasmid expressing human antisense FXYD6. Meanwhile, the empty vector and non-transfection group were designed. The mRNA transcription level of hFXYD6 was assayed by real-time reverse transcriptase polymerase chain reaction （RT-PCR） with SYBR Green I, and the expression of hFXYD6 protein was detected by immunohistochemistry. MTT and the colony-forming assay were used to measure the ability of cell growth. The ceil cycle distribution was analyzed by flow cytometry and Transwell chamber model was employed to test the ability of cell invasion in vitro. RESULTS： In comparison with the cells transfected with empty vector or without transfection, QBC939 cells transfected with antisense hFXYD6 had a significant decrease in mRNA transcription and protein expression. The cell doubling time was augmented （46.8 h vs 34.5 h, 35.3 h）, whereas the colony formation was decreased （24.3% ± 5.3% vs 61.0% ± 8.5%, 58.0% ± 5.6%; P 〈 0.001）. The G1-phase cell population was obviously increased （66.4% ± 2.9% vs 33.5% ± 2.3%, 39.4% ± 3.7%, P 〈 0.001）, but the S-phase one was decreased （18.6% ± 1.6% vs 36.2% ± 2.1%, 34.1% ± 1.6%, P 〈 0.001）. The cells moved from the upper chamber into the lower one in Transwell chamber assay had no marked difference. Between the cells without transfection and transfected with empty vector, there were no significant differences in cell doubling time, colony forming ability, cell cycle distribution and the ability of cell invasion in vitro. CONCLUSION： Transfection of antisense hFXYD6 can inhibit the ability of cell proliferation, but it has no effect on the ability of cell invasion in human cholangiocarcinoma cell line QBC939 in vitro.

关 键 词：FXYD6 反义核酸 胆管癌 增殖 侵袭 荧光定量RT—PCR 免疫组化 流式细胞术 

分 类 号：R735.8[医药卫生—肿瘤]