siRNA干扰Gal-3对Eca-109食管癌细胞株CyclinD1、p21、p27蛋白表达及细胞增殖的影响

Effects of Galectin-3 small interfering RNA on the expressions of Cyclin D1,p21,p27 protein and the effect on cell proliferation of Eca109 esophageal carcinoma cell line

目的探讨siRNA干扰Galectin-3 mRNA后,食管癌Eca-109细胞株增殖活性的影响及Ga-lectin-3、CyclinD1、p21、p27蛋白的表达。方法将Galectin-3-siRNA转染食管癌Eca-109细胞株后,分别用Real-timePCR、MTT、流式细胞术和免疫细胞化学分别检测转染效率、细胞增殖活性、细胞周期分布及Ga-lectin-3、CyclinD1、p21、p27蛋白的表达。结果食管癌Eca-109细胞增殖活性明显减少,主要分布在G1期,Galectin-3和CyclinD1蛋白表达下降,p21和p27蛋白的表达升高。结论运用siRNA可有效抑制Ga-lectin-3蛋白的表达及细胞增殖活性,Galectin-3参与食管癌Eca-109细胞株的增殖与CyclinD1、p21、p27蛋白关系密切,Galectin-3可成为治疗食管癌的靶基因。

Objective To observe the changes of cell proliferation activity and expressions of Galectin-3,CyclinD1,p21 and p27 protein in Eca-109 cell line after transfected by Galectin-3 small interfering RNA（siRNA）.Methods The Galectin-3 siRNA chain was constructed and transfected into Eca-109 cell line by lipofectamine 2000,then Real-time PCR,MTT,Flow cytometry and immuocytochemistry were respectively used to detect the transfection efficiency of siRNA,the cell proliferation activity,cell cycle distribution and the expression levels of Galectin-3,Cyclin D1,p21 and p27protein in Eca-109 cell line.Results The cell proliferation activity of Eca-109 cell line was significantly decreased,and cell distribution was mainly at G1 stage,and the expression levels of Galectin-3 and Cyclin D1 were decreased,however,the expression levels of p21 and p27 protein were increased.Conclusion The application of siRNA can effectively inhibit the expression of Galectin-3 and cell proliferation activity of Eca-109 cell line,and Galectin-3 is involved in the proliferation of Eca-109 cell line and is closely correlated with the expressions of Cyclin D1,p21,p27 protein,which may become the target gene for treating esophageal cancer.