TGF-β1反义寡核苷酸对食管鳞癌细胞EC9706增殖及凋亡的影响  

TGF-β1 antisense oligonucleotide ASODN promotes cell proliferation but inhibits apoptosis in human esophageal squamous cell carcinoma cell line EC9706

作　　者：张红燕[1] 李珊珊[1] 孙洋[1] 王新华[1] 阎爱华[2] 王小军[1] 

机构地区：[1]郑州大学第一附属医院病理科河南省肿瘤病理重点实验室,河南省郑州市450052 [2]郑州大学基础医学院癌前研究室,河南省郑州市450052

出　　处：《世界华人消化杂志》2009年第34期3480-3485,共6页World Chinese Journal of Digestology

基　　金：河南省重点科技攻关基金资助项目;No.072102310042~~

摘　　要：目的:探讨转染TGF-β1反义寡核苷酸(TGF-β1-ASODN)对食管鳞癌细胞EC9706增殖及凋亡的影响.方法:将化学合成的TGF-β1-ASODN转染食管鳞癌细胞EC9706,采用RT-PCR和流式细胞术检测转染效率;观察TGF-β1-ASODN转染后细胞形态学的改变;采用MTT和流式细胞术检测TGF-β1-ASODN转染后对细胞增殖及凋亡的影响.结果:转染TGF-β1-ASODN可有效抑制EC9706细胞中TGF-β1的活性,其mRNA及蛋白的表达均明显低于转染前的水平(0.25±0.07vs0.43±0.09;35.35%vs41.38%,均P<0.05);TGF-β1-ASODN可明显促进细胞增殖、抑制细胞凋亡,转染后细胞生长拥挤失去正常形态,细胞存活率高于转染前(109.4%vs100.0%,P<0.05),G1期、S期细胞百分比低于转染前,G2期细胞百分比则高于转染前,细胞凋亡率低于转染前(62.9%vs66.5%;21.3%vs23.7%;14.8%vs9.8%;0.69%vs0.96%,均P<0.05).结论:TGF-β1-ASODN可高效特异地将TGF-β1基因沉默,并解除其抑制细胞增殖、阻滞细胞周期、促进细胞凋亡的作用.AIM： To investigate the effects of transforming growth factor-beta 1 antisense oligonucleotide ASODN （TGF-~I-ASODN） on cell proliferation and apoptosis in human esophageal squamous cell carcinoma cell line EC9706. METHODS： EC9706 cells were transfected with chemically synthesized TGF-β1-ASODN. The expression of TGF-β1 mRNA and protein was detected by reverse transcription-polymerase chain reaction （RT-PCR） and flow cytometry, respectively. Cell morphological changes were observed. The proliferation and apoptosis of EC9706 cells were measured by methyl thiazolyl tetrazolium （MTT） assay and flow cytometry, respectively. RESULTS： After TGF-β1-ASODN transfec- tion, the expression levels of TGF-β1 mRNA and protein in EC9706 cells were significantly lower than those in untransfected EC9706 cells （0.25 ± 0.07 vs 0.43 ± 0.09 and 35.35% vs 41.38%, respectively; both P 〈 0.05）. TGF-β1-ASODN transfection could stimulate proliferation and inhibit apoptosis of EC9706 cells. After transfec- tion, EC9706 cells lost their normal morphology. Compared with untransfected cells, the survival rate increased （109.4% vs 100.0%, P 〈 0.05）, the percentages of cells in G1 and S phases decreased （62.9% vs 66.5% and 21.3% vs 23.7%, respec- tively; both P 〈 0.05）; the percentage of cells in G2 phase rose （14.8% vs 9.8%, P 〈 0.05）, and the apoptosis rate dedined in cells transfected with TGF-β1-ASODN （0.69% vs 0.96%, P 〈 0.05）. CONCLUSION： TGF-β1-ASODN can silence the expression of the TGF-β1 gene efficiently and specially, and antagonize TGF-β1-rnediated pro- liferation inhibition, cell cycle arrest and apopto- sis in EC9706 cells.

关 键 词：食管鳞癌 转化生长因子Β1 增殖 凋亡 

分 类 号：R735.1[医药卫生—肿瘤] R575.2[医药卫生—临床医学]