Jarid1b对食管鳞癌KYSE-150细胞增殖及分化的影响

EFFECT OF JARID1BON THE PROLIFERATION AND DIFFERENTIATION OF ESOPHAGEAL SQUAMOUS CELL CARCINOMA KYSE-150CELLS

目的研究去甲基化酶Jarid1b对食管鳞癌细胞增殖及分化的影响,为食管鳞癌治疗提供新的靶点。方法采用实时荧光定量PCR(RT-qPCR)以及Western-blot方法检测低分化食管鳞癌细胞系KYSE-150和高分化食管鳞癌细胞系TE-1中去甲基化酶Jarid1b、细胞角蛋白10(CK10)、细胞角蛋白13(CK13)的表达。采用慢病毒转染的方式获得能够在药物诱导时过表达Jarid1b的KYSE-150稳转细胞系,应用CCK8细胞毒性实验检测过表达Jarid1b对该细胞系增殖能力的影响;采用RT-qPCR和Western-blot方法检测过表达Jarid1b后细胞系CK10和CK13表达量的变化。结果 RT-qPCR检测结果表明,和低分化KYSE-150细胞系相比,高分化TE-1细胞系中CK10和CK13高表达(t=22.07、38.44,P<0.05);Western-blot检测结果表明,和KYSE-150细胞系相比,TE-1细胞系中Jarid1b的蛋白表达水平明显增高。CCK8实验结果显示,与未诱导的KYSE-150细胞系相比,诱导KYSE-150细胞系过表达Jarid1b24、48h后,该细胞的增殖能力明显下降(t=10.94、16.71,P<0.05)。RT-qPCR和Western-blot方法检测结果表明,诱导KYSE-150细胞系过表达Jarid1b以后,与未诱导的KYSE-150细胞系相比CK10和CK13的表达明显增高(t=9.706、5.23,P<0.05)。结论去甲基化酶Jarid1b过表达可以抑制低分化食管鳞癌细胞KYSE-150的增殖并促进其分化。

Objective To investigate the effect of Jarid1b,a demethylase,on the proliferation and differentiation of esophageal squamous cell carcinoma（ESCC）cells,and to provide a new target for the treatment of ESCC. Methods RT-qPCR and Western-blot were used to measure the expression of Jarid1b,cytokeratin 10（CK10）,and cytokeratin 13（CK13）in poorly differentiated ESCC cell line KYSE-150 and well-differentiated ESCC cell line TE-1.KYSE-150 cells with stable overexpression of Jarid1b when induced by drugs were obtained by lentivirus transfection.CCK-8 cytotoxicity assay was used to analyze the effect of Jarid1b overexpression on the proliferative capacity of this cell line,and RT-qPCR and Western blot were used to measure the changes in the expression of CK10 and CK13 in this cell line after Jarid1b overexpression. Results RT-qPCR data showed that welldifferentiated TE-1 cells had higher expression of CK10 and CK13 than poorly differentiated KYSE-150 cells（t=22.07,38.44,P〈0.05）.Western blot showed that compared with KYSE-150 cells,TE-1 cells had a significant increase in the protein expression of Jarid1b.The CCK-8 assay showed that compared with the uninduced KYSE-150 cells,the induced KYSE-150 cells had a significant reduction in proliferative capacity at 24 and 48 hours after Jarid1b overexpression（t=10.94,16.71,P〈0.05）.RT-qPCR and Western blot showed that after Jarid1b overexpression,the induced KYSE-150 cells had significant increases in the expression of CK10 and CK13 compared with the uninduced KYSE-150 cells（t=9.706,5.23,P〈0.05）. Conclusion Overexpression of the demethylase Jarid1b can inhibit the proliferation and promote the differentiation of poorly differentiated ESCC KYSE-150 cells.