收稿日期: 2016-04-09
网络出版日期: 2018-03-05
基金资助
中国科学院``百人计划 A 类''资助项目(Y5BES11001)
Effects of ABL silencing combined with TRAIL and DOX on apoptosis of colorectal cancer HT29 cells
Received date: 2016-04-09
Online published: 2018-03-05
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.
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
李雨雨, 金由辛, 徐中娟, 张书忙, 索广力 . ABL 沉默对 DOX 和 TRAIL 诱导结肠癌细胞 HT29 凋亡的影响[J]. 上海大学学报(自然科学版), 2018 , 24(1) : 134 -141 . DOI: 10.12066/j.issn.1007-2861.1771
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.
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