Position: Home > Articles > Construction of Prx Ⅱ Knock-down L02 Cell line and Effect on Proliferation
Journal of Heilongjiang Bayi Agricultural University
2019
(1)
49-55
PrxⅡ基因敲降L02细胞系的构建及其对细胞增殖的影响
作 者：
满艺;甄兴;孔令祖;申贵男;韩英浩;金美华
单 位：
黑龙江八一农垦大学生命科学技术学院
关键词：
L02细胞系;PrxⅡ;细胞增殖;RNA干扰技术;慢病毒
摘 要：
构建PrxⅡ基因敲降L02细胞系并确定PrxⅡ基因对L02细胞增殖的影响。利用慢病毒介导的shRNA干扰技术、流式细胞术、荧光显微照相、蛋白质免疫印迹、MTT asssay等方法建立并检验PrxⅡ敲降L02细胞系及其对L02细胞的增殖影响。结果显示:选用5’-CCGCTTGTCTGAGGATACGTT-3’片段作为干扰PrxⅡ基因片段,利用慢病毒敲降L02细胞。同时处理嘌呤霉素筛选得到shPrxⅡ组细胞和Scramble组细胞,使用流式细胞术、荧光显微照相技术对shPrxⅡ组细胞和Scramble组细胞进行检测,结果均显示出GFP蛋白表达;此后利用蛋白质免疫印迹法检测出shPrxⅡ组细胞中PrxⅡ蛋白表达水平较Blank、Scram-ble组显著降低(P<0.05),Blank组与Scramble组细胞中PrxⅡ蛋白表达水平无明显差异。在MTT结果中显示shPrxⅡ组细胞增殖速率明显高于Scramble组,且具有统计学意义。成功构建PrxⅡ敲降的L02细胞系,发现PrxⅡ敲降后显著提高L02细胞的增殖速率。
译 名：
Construction of Prx Ⅱ Knock-down L02 Cell line and Effect on Proliferation
作 者：
Man Yi;Zhen Xing;Kong Lingzu;Shen Guinan;Han Yinghao;Jin Meihua;College of Life Science and Technology,Heilongjiang Bayi Agricultural University;
单 位：
Man Yi%Zhen Xing%Kong Lingzu%Shen Guinan%Han Yinghao%Jin Meihua%College of Life Science and Technology,Heilongjiang Bayi Agricultural University
关键词：
L02 cell line;;PrxⅡ;;cell proliferation;;RNA interference technology;;lentivirus
摘 要：
To construct PrxⅡ gene knock-down L02 cell line and determine the effect of PrxⅡ gene on L02 cell proliferation,the expression of PrxⅡ gene was knockdown by lentivirus-mediated shRNA interference,the transfection of lentivirus was detected by flow cytometry and fluorescence microscopy,the knockdown efficiency of PrxⅡ was detected by Western blot,and the effect of PrxⅡknockdown on the proliferation of L02 cells examined by MTT assay.The sequence 5'CCGCTTGTCTGAGGATACGTT-3 'was selected as the fragment of interfering PrxⅡ gene,and the cell of shPrxⅡ and Scramble groups were screened by puromycin treatment.The result detected by flow cytometry and fluorescence microscopy showed that GFP protein expressed.The expression of PrxⅡprotein in shPrxⅡ group was significantly lower than that in Blank group and Scramble group(P <0.05).PrxⅡ expression had no significant difference between Blank group and Scramble group.MTT result showed that cell proliferation rate of shPrxⅡ group was significantly higher than the Scramble group,and with statistical significance.PrxII gene knock-down L02 cell line was successfully constructed and PrxII knock-down significantly increased the proliferation rate of L02 cells.
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