CHAC1基因沉默SU-DHL-8细胞株的构建及其对青蒿素细胞毒活性的影响

Construction of SU-DHL-8 cell line with silenced CHAC1 gene and its effect on cytotoxic activity of artemisinin

应用慢病毒短发夹RNA(short hairpin RNA,shRNA)转染的方式构建了阳离子转运蛋白样蛋白1(cation transport regulator-like protein 1,CHAC1)基因沉默SU-DHL-8细胞株模型,用于研究CHAC1基因对青蒿素细胞毒活性的影响.研究了青蒿素对弥散性大B细胞淋巴瘤SU-DHL-8细胞增殖的抑制作用及作用后CHAC 1基因及蛋白的表达情况.应用小干扰RNA(small interfering RNA,siRNA)沉默SU-DHL-8细胞株内的CHAC1基因.采用慢病毒转染的方法构建CHAC1基因沉默的SU-DHL-8细胞株.采用实时荧光定量核酸扩增检测系统(real-time quantitative PCR detecting system,qPCR)、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT)活性测定法和蛋白免疫印迹(Westen blot)的方法检测沉默效果以及CHAC1基因对青蒿素细胞毒活性的影响.成功构建了稳定沉默CHAC1基因的稳转细胞株,并初步探讨了CHAC1基因对青蒿素抑制弥散性大B细胞淋巴瘤SU-DHL-8细胞增殖的影响.

To study the effect of cation transport regulator-like protein 1(CHAC 1)gene on the cytotoxic activity of artemisinin,the SU-DHL-8 cell line with silenced CHAC 1 gene was constructed by using lentiviral shRNA transfection.The inhibitory effect of artemisinin on the proliferation of lymphoma cell line SU-DHL-8 and the expression of CHAC 1 gene and protein were studies.Small interfering RNA(siRNA)was used to silence CHAC 1 gene in SU-DHL-8 cell line.The SU-DHL-8 cell line with silenced CHAC 1 gene was constructed by lentiviral transfection.The Real-time Quantitative PCR Detecting System(qPCR),3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)activity assay and Western blot were used to detect the silencing effect and the effect of CHAC 1 gene on the cytotoxic activity of artemisinin.A stable transfected SU-DHL-8 cell line with stable silenced CHAC 1 gene was successfully constructed,and the effect of CHAC 1 gene on the proliferation of lymphoma cell line SU-DHL-8 inhibited by artemisinin was preliminarily investigated.