滋肾固髓汤通过调控p38MAPK/p53信号通路诱导结直肠癌HCT-116细胞凋亡  

Zishen Gusui Decoction induces apoptosis of colorectal cancer HCT-116 cells by regulating p38MAPK/p53 signaling pathway

作　　者：吴亭[1] 张云芳[1] 邱晓堂[1] 陈学武[1] 姜靖雯[1] 邢明远 苏乙花 WU Ting;ZHANG Yunfang;QIU Xiaotang;CHEN Xuewu;JIANG Jingwen;XING Mingyuan;SU Yihua(Department of Oncology,Hainan Hospital of Traditional Chinese Medicine,Hainan Haikou 570203,China)

机构地区：[1]海南省中医院肿瘤科,海南海口570203

出　　处：《现代肿瘤医学》2023年第9期1608-1613,共6页Journal of Modern Oncology

基　　金：海南省自然科学基金委员会基础与应用基础研究计划(编号:821MS0837)。

摘　　要：目的:探讨滋肾固髓汤对结直肠癌HCT-116细胞凋亡的影响及其分子机制。方法:制备滋肾固髓汤含药血清,首先采用不同体积分数滋肾固髓汤含药血清处理HCT-116细胞,噻唑蓝(MTT)检测细胞增殖活性;再将HCT-116细胞分为空白组、滋肾固髓汤低、中、高剂量组、SB203580(p38MAPK抑制剂)组和高剂量滋肾固髓汤+SB203580组,分别加入相应药物进行干预后,Hoechst 33258染色观察细胞凋亡的形态学变化;Annexin V-FITC流式细胞术检测细胞的凋亡水平;Western blot检测细胞中p38MAPK、磷酸化(p)-p38MAPK(Thr180/Tyr182)、鼠双微染色体2(MDM2)、p53、Cleaved caspase-3、Bax和Bcl-2等蛋白表达水平。结果:滋肾固髓汤可抑制HCT-116细胞增殖活性,并呈时间和浓度依赖性。不同浓度滋肾固髓汤处理可诱导HCT-116细胞呈现核聚集、皱缩或碎裂等凋亡形态变化,促进细胞凋亡,上调p-p38MAPK、p53、Bax和Cleaved caspase-3等蛋白表达水平,下调MDM2和Bcl-2蛋白表达水平。SB203580抑制滋肾固髓汤诱导HCT-116细胞凋亡及p38MAPK/p53信号通路活化的作用。结论:滋肾固髓汤通过激活p38MAPK/p53信号通路诱导人结直肠癌HCT-116细胞凋亡。Objective:To investigate the effect of Zishen Gusui Decoction on apoptosis of colorectal cancer HCT-116 cells and its molecular mechanism.Methods:Zishen Gushu Decoction containing serum was prepared.Firstly,HCT-116 cells were treated with different volume fraction of Zishen Gusui Decoction,and the proliferation activity of HCT-116 cells was detected by methyl thiazolyl-tetrazolium(MTT).Then HCT-116 cells were divided into blank group,Zishen Gushu Decoction low,medium and high dose groups,SB203580(p38MAPK inhibitor)group and Zishen Gushu Decoction high dose+SB203580 group.After the intervention with the corresponding drugs,Hoechst 33258 staining was used to observe the morphological changes of cell apoptosis.Annexin V-FITC flow cytometry was used to determine the apoptosis rate of the cells.Western blot was used to detect the protein expression levels of p38MAPK,phosphorylated(p)-p38MAPK(Thr180/Tyr182),mouse dimicrosomal 2(MDM2),p53,Cleaved caspase-3,Bax and Bcl-2.Results:Zishen Gusui Decoction could inhibit the proliferation of HCT-116 cells in a time-dependent and concentration-dependent manner.Treatment with different concentrations of Zishen Gusui Decoction could induce apoptotic morphological changes of HCT-116 cells such as nuclear aggregation,shrinkage or fragmentation,promote cell apoptosis,up-regulate the protein expression levels of p-p38MAPK,p53,Bax and Cleaved caspase-3 and down-regulate the protein expression levels of MDM2 and Bcl-2.However,SB203580 inhibited the apoptosis of HCT-116 cells and the activation of p38MAPK/p53 signaling pathway induced by Zishen Gushu Decoction.Conclusion:Zishen Gusui Decoction can induce apoptosis of HCT-116 cells by activating p38MAPK/p53 signaling pathway.

关 键 词：滋肾固髓汤 结直肠癌 细胞凋亡 p38MAPK/p53信号通路 

分 类 号：R735.35[医药卫生—肿瘤]