中国肿瘤生物治疗杂志
中國腫瘤生物治療雜誌
중국종류생물치료잡지
CHINESE JOURNAL OF CANCER BIOTHERAPY
2010年
1期
67-70
,共4页
王桂华%罗学来%孙黎%邓豫%王珅%李兆明%李小兰%陶德定%胡俊波%龚建平
王桂華%囉學來%孫黎%鄧豫%王珅%李兆明%李小蘭%陶德定%鬍俊波%龔建平
왕계화%라학래%손려%산예%왕신%리조명%리소란%도덕정%호준파%공건평
结肠肿瘤%GRIM-19%凋亡%Annexin-V/PI%线粒体膜电位
結腸腫瘤%GRIM-19%凋亡%Annexin-V/PI%線粒體膜電位
결장종류%GRIM-19%조망%Annexin-V/PI%선립체막전위
colon cancer%GRIM-19%apoptosis%Annexin-V/PI%mitochondrial membrane potential
目的:研究干扰素/维甲酸诱导死亡基因(retinoid-interferon-induced mortality,GRIM-19)对结肠癌SW480细胞凋亡的影响.方法:构建GRIM-19真核表达载体(pCMV-Flag-GRIM-19),转染入SW480细胞中,Western blotting检测GRIM-19及凋亡相关蛋白Bal-xl的表达,采用Annexin-V/PI和线粒体膜电位JC-1染色检测SW480细胞的凋亡.结果:成功构建pCMV-Flag-GRIM-19真核表达载体.pCMV-Flag-GRIM-19质粒转染SW480细胞后,GRIM-19的表达上调,凋亡抑制蛋白Bcl-xl的表达则下调.转染空质粒pCMV-Flag组SW480细胞凋亡率为(7.7±1.39)%,转染pCMV-Flag-GRIM-19质粒后SW480细胞凋亡率为(15.0±2.52)%(P<0.05).线粒体膜电位检测显示转染空质粒pCMV-Flag组SW480细胞膜电位降低细胞为(7.5±2.09)%,而转染pCMV-Flag-GRIM-19后细胞线粒体膜电位降低细胞为(17.5±3.07)%(P<0.05).结论:GRIM-19体外转染可有效诱导结肠癌SW480细胞凋亡.
目的:研究榦擾素/維甲痠誘導死亡基因(retinoid-interferon-induced mortality,GRIM-19)對結腸癌SW480細胞凋亡的影響.方法:構建GRIM-19真覈錶達載體(pCMV-Flag-GRIM-19),轉染入SW480細胞中,Western blotting檢測GRIM-19及凋亡相關蛋白Bal-xl的錶達,採用Annexin-V/PI和線粒體膜電位JC-1染色檢測SW480細胞的凋亡.結果:成功構建pCMV-Flag-GRIM-19真覈錶達載體.pCMV-Flag-GRIM-19質粒轉染SW480細胞後,GRIM-19的錶達上調,凋亡抑製蛋白Bcl-xl的錶達則下調.轉染空質粒pCMV-Flag組SW480細胞凋亡率為(7.7±1.39)%,轉染pCMV-Flag-GRIM-19質粒後SW480細胞凋亡率為(15.0±2.52)%(P<0.05).線粒體膜電位檢測顯示轉染空質粒pCMV-Flag組SW480細胞膜電位降低細胞為(7.5±2.09)%,而轉染pCMV-Flag-GRIM-19後細胞線粒體膜電位降低細胞為(17.5±3.07)%(P<0.05).結論:GRIM-19體外轉染可有效誘導結腸癌SW480細胞凋亡.
목적:연구간우소/유갑산유도사망기인(retinoid-interferon-induced mortality,GRIM-19)대결장암SW480세포조망적영향.방법:구건GRIM-19진핵표체재체(pCMV-Flag-GRIM-19),전염입SW480세포중,Western blotting검측GRIM-19급조망상관단백Bal-xl적표체,채용Annexin-V/PI화선립체막전위JC-1염색검측SW480세포적조망.결과:성공구건pCMV-Flag-GRIM-19진핵표체재체.pCMV-Flag-GRIM-19질립전염SW480세포후,GRIM-19적표체상조,조망억제단백Bcl-xl적표체칙하조.전염공질립pCMV-Flag조SW480세포조망솔위(7.7±1.39)%,전염pCMV-Flag-GRIM-19질립후SW480세포조망솔위(15.0±2.52)%(P<0.05).선립체막전위검측현시전염공질립pCMV-Flag조SW480세포막전위강저세포위(7.5±2.09)%,이전염pCMV-Flag-GRIM-19후세포선립체막전위강저세포위(17.5±3.07)%(P<0.05).결론:GRIM-19체외전염가유효유도결장암SW480세포조망.
Objective: To investigate the effect of retinoid-interferon-induced mortality (GRIM-19) gene on the apoptosis of colon cancer. Methods: A GRIM-19 eukaryotic expression vector (pCMV-Flag-GRIM-19) was constructed and transfected into SW480 cells. Expressions of GRIM-19 and apoptosis-related proteins were detected by Western blotting analysis. Apoptosis of SW480 cells was measured by Annexin-V/PI assay and mitochondrial membrane potential JC-1 staining. Results: The GRIM-19 eukaryotic expression vector pCMV-Flag-GRIM-19 was successfully constructed. Expression of GRIM-19 in SW480 cells was up-regulated and that of apoptosis-related protein Bcl-xl was down-regulated after transfection with pCMV-Flag-GRIM-19. Apoptosis rate was (7.7±1.39)% in SW480 cells transfected with pCMV-Flag empty vector and (15.0 ± 2.52)% in pCMV-Flag-GRIM-19 transfected cells (P<0.05). Mitochondrial membrane potential was decreased in (7.5±2.09)% of pCMV-Flag transfected cells and (17.5±3.07)% of pCMV-Flag-GRIM-19 transfected cells (P<0.05). Conclusion: In vitro GRIM-19 transfection can effectively induce apoptosis of colon cancer SW480 cells.
