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抑制JTV1基因表达对大黄素引起的K562细胞凋亡的影响及其机制

ISSN号：1004-5503
期刊名称：中国生物制品学杂志
时间：2011
页码：634-638
分类：Q255[生物学—细胞生物学]
作者机构：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016
相关基金：国家自然科学基金（30300449）; 国家中医药管理局（02-03ZP52）; 重庆医科大学课题（XBYB2007104）
相关项目：APL诊疗新靶点的筛选与研究

关键词： JTV1基因, K562细胞, 细胞凋亡, BCL-2基因, C-MYC基因, BAX基因, JTV1 gene, K562 cells, Cell apoptosis, Bcl-2 gene, C-myc gene, Bax gene

中文摘要：

目的探讨抑制JTV1基因表达对大黄素引起的人白血病K562细胞系凋亡的影响及其机制。方法将pGene-Sil-1-JTV1-1.1 siRNA重组质粒、pGeneSil-1-N.1阴性对照质粒及pGeneSil-1空载体分别转染人K562细胞,通过集落形成试验检测细胞的增殖能力;各组转染K562细胞经80μmol/L大黄素处理后,流式细胞术分析细胞周期和细胞凋亡率,RT-PCR法检测细胞中凋亡相关基因Bcl-2、Bax、C-myc转录水平的变化,Western blot法检测Bcl-2、Bax、C-myc翻译水平的变化。结果抑制JTV1基因的表达后,K562细胞的增殖能力明显提高;抑制JTV1基因表达可使经80μmol/L大黄素处理的K562细胞G1期细胞比例下降,并减轻大黄素引起的细胞凋亡,同时,细胞Bax基因mRNA的转录水平和蛋白表达水平均明显下降,而Bcl-2基因和C-myc基因mRNA的转录水平和蛋白表达水平则明显上调。结论抑制JTV1基因表达可能通过上调Bcl-2和C-myc基因的表达,下调Bax基因的表达,促进K562细胞的增殖,并阻止细胞凋亡。

英文摘要：

Objective To investigate the effect of inhibiting expression of tumor-suppressing gene JTV1 on the emodin-induced apoptosis of human chronic myeloid 1eukemia（CML） K562 cell line as well as the relevant mechanism.Methods K562 cells were transfected with recombinant plasmids pGeneSil-1-JTV1-1.1 siRNA,pGeneSil-1-N.1 as negative control and empty vector pGeneSil-1 respectively and determined for proliferation level by colony formation test.The transfected K562 cells in various groups were treated with 80 μmol/L emodin for 72 h,then analyzed for cell cycle and apoptosis rate by flow cytometry,for transcription levels of apoptosis-related Bax,Bcl-2 and C-myc genes by RT-PCR,and for translation levels of the genes by Western blot.Results After the expression of JTV1 gene was inhibited,the proliferation level of K562 cells increased significantly,the percentage of K562 cells treated with 80 μmol/L emodin at G1 phase decreased,while the emodin-induced apoptosis was relived.Meanwhile,both the mRNA transcription and protein expression levels of Bax gene decreased significantly,while those of Bcl-2 and C-myc genes increased significantly.Conclusion The inhibition of JTV1 gene expression might promoted the proliferation and arrest the apoptosis of K562 cells by up-regulating the expressions of Bcl-2 and C-myc genes and down-regulating the expression of Bax gene.

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