ABL沉默对DOX和TRAIL诱导结肠癌细胞HT29凋亡的影响  

Effects of ABL silencing combined with TRAIL and DOX on apoptosis of colorectal cancer HT29 cells

作　　者：李雨雨 金由辛[1] 徐中娟 张书忙 索广力[2] 

机构地区：[1]上海大学生命科学学院,上海200444 [2]中国科学院苏州纳米技术与纳米仿生研究所,江苏苏州215123

出　　处：《上海大学学报（自然科学版）》2018年第1期134-141,共8页Journal of Shanghai University:Natural Science Edition

基　　金：中国科学院"百人计划A类"资助项目(Y5BES11001)

摘　　要：研究酪氨酸激酶Abelson(ABL)基因沉默与肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis inducing ligand,TRAIL)、阿霉素(doxorubicin,DOX)联合作用对结肠癌细胞株HT29凋亡的影响.利用shRNA慢病毒载体构建ABL沉默稳转细胞株,并用Western blot检测ABL表达水平;细胞计数试剂盒8(cell count kit 8,CCK8)实验检测细胞增殖和药物毒性;流式细胞仪检测细胞凋亡情况;Western blot检测凋亡相关蛋白PARP1,cleaved-caspase3的表达水平.结果表明:TRAIL,DOX均能够抑制结肠癌细胞的细胞活性,二者联合用药可在较低剂量诱导细胞凋亡;ABL基因沉默能够抑制结肠癌细胞增殖,但同时也抑制了TRAIL对HT29细胞的凋亡诱导作用,以及TRAIL和DOX对HT29细胞凋亡的协同作用.可见,DOX和TRAIL联合用药能够显著诱导HT29细胞发生凋亡;虽然ABL沉默能抑制HT29细胞增殖,但同时也抑制了TRAIL和DOX协同促进HT29细胞凋亡的作用.To investigate the effects of Abelson （ABL） tyrosine kinase silencing combined with tumor necrosis factor related apoptosis-inducing ligand （TRAIL） and doxorubicin （DOX） on apoptosis of human colon cancer cell line HT29. ABL was stably knocked down in HT29 cells using lent ivirus particles. The expression level was verified by Western blot. Cell proliferation and drug toxicity was detected by cell count kit 8 （CCK8） assay. Cell apoptosis was detected by flow cytometry. The expression level PARP1 and cleaved- caspase3 were determined by Western blot. The results showed that cell viability was reduced after treatment of TRAIL or DOX. Combination therapy can induce apoptosis rate at a lower dose. The colon cancer cell proliferation was inhibited by ABL gene silencing. However, apoptosis induced by TRAIL was inhibited, and the synergistic apoptotic effect of TRAIL and DOX on HT29 cell was also inhibited. Combination treatment can significantly induce HT29 cell apoptosis. Although ABL silencing can inhibit cell proliferation, the synergistic apoptotic effect of TRAIL and DOX on colorectal cancer HT29 cell was still inhibited by ABL silencing.

关 键 词：结肠癌 酪氨酸激酶Abelson(ABL) 基因沉默 肿瘤坏死因子相关凋亡诱导配体 阿霉素 细胞凋亡 

分 类 号：Q26[生物学—细胞生物学]