基础医学与临床2011，Vol.31Issue(6)：602-608,7.
Haxl基因及其特异性小干扰RNA对人胚肾293细胞凋亡的影响
The effect of Haxl gene and its specific small interference RNAs on the apoptosis of 293 cells
摘要
Abstract
Objective To construct the Haxl gene specific siRNA (sihax) for detection of sihax mediated Haxl gene repression and to study the influence of Haxl and Haxl siRNAs on cell apoptosis. Methods The oligo sequences of Haxl siRNAs were selected, and the constructed eukaryotic expression vectors were transfected into 293 cells. The influence of Haxl and its siRNAs on cell apoptosis was evaluated by a number of approaches such as inverted fluorescence microscope, flow cytometric analysis, annexin V-EGFP/PI double staining, and standard RTPCR. Results Haxl specific siRNAs, pBS/U6-sihaxA and pBS/U6-sihaxB, were successfully constructed. Analysis by RT-PCR and inverted fluorescence microscopy demonstrated that the inhibitory effect induced by sihaxB was stronger than that of sihaxA. Over-expression of Haxl significantly inhibited the early apoptosis of 293 cells. Inhibiting the endogenous Haxl gene expression by by sihaxA and sihaxB markedly increased late apoptotic cells from 9. 03％ ±0. 473％ to 10. 8％ ±0. 513％ (P ＜0. 05) and 16. 6％ ±0. 858％ (P ＜0. 01 ), respectively. In the same time, sibaxB also reduced the early apoptotic cells from 37. 3％ ± 0. 5％ to 32％ ± 1.77％ ( P ＜ 0. 01 ). Conclusion The sequence conser ration of the targeting site may influence the inhibitory effect induced by Haxl siRNA. Haxl sene might actively involve in the multiple regulatory processes related to cell apoptosis. This study provides some valuable information for the functional study on Haxl gene in the future.关键词
Haxl基因/小干扰RNA/293细胞/凋亡Key words
Haxl/ siRNA/ 293 cells/ apoptosis分类
医药卫生引用本文复制引用
龙琳,刘力..Haxl基因及其特异性小干扰RNA对人胚肾293细胞凋亡的影响[J].基础医学与临床,2011,31(6):602-608,7.基金项目
国家自然科学基金(30871283) （30871283）
2010年中国医学科学院基础医学研究所所院长基金(2010PYZ17) （2010PYZ17）
