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 |
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引用本文: | 王宁,杨春菊,何丽囡,樊保敏,曾广智,尹俊林.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[J].云南民族大学学报(自然科学版),2019(4):313-319. |
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作者姓名: | 王宁 杨春菊 何丽囡 樊保敏 曾广智 尹俊林 |
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作者单位: | 云南民族大学云南民族大学-香港浸会大学传统天然药物研发联合实验室 |
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摘 要: | 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CHAC1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.
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关 键 词: | 弥散性大B细胞淋巴瘤 青蒿素 CHAC1 转染 基因沉默 |
Construction of SU-DHL-8 cell line with silenced CHAC1 gene and its effect on cytotoxic activity of artemisinin |
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Institution: | ,YMU-HKBU Joint Laboratory of Traditional Natural Medicine, Yunnan Minzu University |
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Abstract: | To study the effect of cation transport regulator-like protein 1(CHAC1) gene on the cytotoxic activity of artemisinin, the SU-DHL-8 cell line with silenced CHAC1 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 CHAC1 gene and protein were studies. Small interfering RNA(siRNA) was used to silence CHAC1 gene in SU-DHL-8 cell line. The SU-DHL-8 cell line with silenced CHAC1 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 CHAC1 gene on the cytotoxic activity of artemisinin. A stable transfected SU-DHL-8 cell line with stable silenced CHAC1 gene was successfully constructed, and the effect of CHAC1 gene on the proliferation of lymphoma cell line SU-DHL-8 inhibited by artemisinin was preliminarily investigated. |
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Keywords: | diffuse large B-cell lymphoma artemisinin CHAC1 transfection gene silencing |
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