敲低DR5表达对rmhTRAIL联合5-FU干预结肠癌细胞增殖及凋亡的影响  

Effects of DR5 knockdown on proliferation and apoptosis of human colorectal cancer cells intervened by rmhTRAIL combined with 5-FU

作　　者：孙同友[1] 周硕 梁秀军[3] 武慧杰 廉蕊[1] SUN Tongyou;ZHOU Shuo;LIANG Xiujun;WU Huijie;LIAN Rui(Department of Radiotherapy and Chemotherapy,Chengde Central Hospital,Chengde 067000,China)

机构地区：[1]承德市中心医院放化疗科,河北承德067000 [2]承德医学院研究生院 [3]承德医学院基础医学研究所

出　　处：《山东医药》2022年第10期6-9,14,共5页Shandong Medical Journal

基　　金：河北省卫生健康委科研基金项目(20211025);河北省教育厅科技项目(QN2016031);承德市科技计划项目(201804A078)。

摘　　要：目的探讨敲低死亡受体5(DR5)表达对重组变构人肿瘤坏死因子相关凋亡诱导配体(rmhTRAIL)联合5氟尿嘧啶(5-FU)抑制人结直肠癌(CRC)HCT116细胞增殖及凋亡的影响。方法体外培养人CRC HCT116细胞,用DR5 shRNA质粒转染,构建敲低DR5的CRC HCT116/shRNA细胞系(HCT116/KD);MTS法检测rmhTRAIL、5-FU及两者联用对HCT116/KD细胞增殖的影响;流式细胞术检测细胞凋亡;Western blotting法检测细胞凋亡信号通路中凋亡相关蛋白Caspase-3、Caspase-8、Caspase-9、PARP和PARP裂解片段表达。结果成功构建敲低DR5的CRC HCT116/KD细胞系;MTS及流式细胞术结果显示,敲低DR5表达后,rmhTRAIL单药及与5-FU联用对CRC HCT116细胞的增殖抑制及凋亡诱导作用减弱;Western blotting结果显示,rmhTRAIL单药及其联合5-FU作用于HCT116/KD细胞,Caspase-3、Caspase-9和PARP表达未见明显下降(P均>0.05)。结论敲低DR5表达后rmhTRAIL单药及其联合5-FU干预HCT116细胞,细胞增殖受抑减弱,凋亡减少。Objective To investigate the effects of knockdown of death receptor 5(DR5)expression on the prolifera⁃tion and apoptosis of human colorectal cancer(CRC)HCT116 cells intervened by recombinant mutant human tumor necro⁃sis factor-related apoptosis-inducing ligand(rmhTRAIL)combined with 5-fluorouracil(5-FU).Methods Human CRC HCT116 cells were cultured in vitro and transfected with DR5 shRNA plasmid to construct CRC HCT116/shRNA cell line(HCT116/KD)with DR5 knockdown.MTS assay was used to detect the effects of rmhTRAIL,5-FU and their combination on the proliferation of HCT116/KD cells.Flow cytometry was used to detect their apoptosis.The expression levels of apop⁃tosis signaling pathway-related proteins Caspase-3,Caspase-8,Caspase-9,poly-ADP-ribose polymerase(PARP)and PARP cleavage fragments were detected by Western blotting.Results CRC HCT116/KD cell line was successfully con⁃structed.The results of MTS and flow cytometry showed that the proliferation inhibition and apoptosis induction of CRC HCT116 cells treated by rmhTRAIL and its combination with 5-FU were weakened after DR5 was knocked down.Western blotting showed that the expression levels of Caspase-3,Caspase-9 and PARP did not decrease significantly in CRC HCT116/KD cells treated with rmhTRAIL or rmhTRAIL in combination with 5-FU(all P>0.05).Conclusions When HCT116 cells are treated with rmhTRAIL or rmhTRAIL combined with 5-FU,the cell proliferation inhibition is weakend and apoptosis is reduced after DR5 is knocked down.

关 键 词：结直肠癌 肿瘤坏死因子相关凋亡诱导配体 细胞凋亡 5-氟尿嘧啶 死亡受体 

分 类 号：R73.3[医药卫生—肿瘤]