【摘要】 目的通过RT-PCR方法观察TGF-β1在大肠癌组织中的mRNA表达及RNA干扰阻断TGF-β1表达对大肠癌细胞生长周期和侵袭力的影响。方法收集20例各期大肠癌手术标本抽提RNA,用半定量RT-PCR方法检测各组织中TGF-β1的mRNA表达。用小片段TGF-β1的双链RNA(siRNA)转染大肠癌HCT116细胞,半定量RT-PCR检测TGF-β1mRNA受抑程度,流式细胞仪观察转染后细胞周期变化,软琼脂克隆实验检测肿瘤细胞生长及成瘤性的变化。结果在20例各期大肠癌中TGF-β1检测12例(60%)存在mRNA过度表达。小片断RNA转染显著抑制了肿瘤细胞TGF-β1的表达,使细胞HCT 116的S期细胞较对照组明显增高59%~82.5%,进入G2期细胞减少;转染组细胞克隆形成率较对照组显著下降。结论TGF-β1mRNA在大肠癌组织中过度表达,通过RNA干扰技术可以快捷方便的阻断TGF-β1mRNA表达,使大肠癌细胞S期阻滞并抑制肿瘤细胞生长、降低其成瘤性。更多还原

【Abstract】 Objective To observe TGF-β1 mRNA expression in the tissue of colon cancer and the effects of inhibition of TGF-β1 on the growth and aggression of colon cancer cells.Methods The specimens of 20 patients with colon cancer were collected and total RNA was extracted for RT-PCR detection.HCT 116 colon cancer cells were transfected with small interfering dsRNAs(siRNA).The effects of the inhibition of TGF-β1 were tested by semi-quantitative RT-PCR and changes in cell cycle distribution were analyzed by flow cytometry,while the ability of aggression was detected by soft agar colony assay.Results The mRNA expressions of TGF-β1 were over-expressed in 12/20(60%) colon cancer tissues.The inhibition of the expression of TGF-β1 was significantly different after the transfection of siRNA against TGF-β1,and this induced the increase in S phase by 59%～82.5% and decrease in G2 phase,colony assay further demonstrated that siRNA against TGF-β1 led to a significant reduction in colony formation as compared with that of the control group.Conclusion The results suggest that mRNA over-expression of TGF-β1 is found in patients with colon cancer.And mRNA expression of TGF-β1 in the colon cancer cells is conveniently blocked by siRNA transfection.更多还原