茯苓酸抑制人结肠癌细胞系HT-29增殖  

Pachymic acid inhibits proliferation of human colon cancer cell line HT-29

作　　者：冯娜欣 王鹏[1] 何明璇 刘锦燕[1] FENG Naxin;WANG Peng;HE Mingxuan;LIU Jinyan(Department of Oncology,the No.2 Hospital of Baoding,Baoding 071051,China)

机构地区：[1]保定市第二医院肿瘤内科,河北保定071051

出　　处：《基础医学与临床》2023年第11期1673-1679,共7页Basic and Clinical Medicine

基　　金：保定市科技计划(2041ZF205)。

摘　　要：目的探讨茯苓酸(PA)对人结肠癌细胞系HT-29增殖和凋亡的影响及可能的分子机制。方法将细胞分为空白组、20、40和80μmol/L PA组。MTT法检测HT-29细胞增殖;流式细胞仪测定HT-29细胞周期和凋亡;Western blot检测HT-29细胞中细胞周期、凋亡以及Akt/MDM2/p53信号通路相关蛋白质表达。构建HT-29细胞的肿瘤异种移植模型,随后将成瘤裸鼠随机分为对照组、腹腔注射10、30和60 mg/kg PA组(每组6只),测量肿瘤体积、质量;免疫组化测定Ki-67、cleaved caspase-3蛋白表达;Western blot检测肿瘤组织中Akt/MDM2/p53信号通路相关蛋白表达。结果各剂量PA均可呈浓度依赖性显著降低HT-29细胞增殖活力,S、G_(2)/M期细胞比例,细胞中B细胞淋巴瘤基因-2(Bcl-2)、细胞周期蛋白D1(cyclin D1)、细胞周期蛋白依赖性激酶2(CDK2)蛋白表达水平和p-Akt/Akt、p-MDM2/MDM2比值(P<0.05),同时升高细胞凋亡率、G_(0)/G_(1)期细胞比例以及细胞中Bcl-2相关X蛋白(Bax)、切割型胱天蛋白酶-3(cleaved caspase-3)、p53蛋白表达水平,差异均有统计学意义(P<0.05)。各剂量PA均可显著降低裸鼠肿瘤体积、质量以及肿瘤组织中增殖细胞核抗原(Ki-67)蛋白表达水平和p-Akt/Akt、p-MDM2/MDM2比值(P<0.05),同时升高肿瘤组织中cleaved caspase-3、p53蛋白表达水平,差异均有统计学意义(P<0.05)。结论PA可诱导结肠癌细胞凋亡和细胞周期阻滞并抑制细胞增殖,其机制可能与调节Akt/MDM2/p53信号通路有关。Objective To investigate the impacts of pachymic acid(PA)on proliferation and apoptosis of human colon cancer cell line(HT-29)and its potential mechanism.Methods The cells were divided into blank group,20,40 and 80μmol/L PA groups.The proliferation of HT-29 cells was detected by MTT method;The cell cycle and apoptosis of HT-29 cells were detected by flow cytometry;The expression of cell cycle,apoptosis and Akt/MDM2/p53 signal pathway related proteins in HT-29 cells were detected by Western blot.The tumor xeno-transplantation model with HT-29 cells was constructed,and then the tumorigenic nude mice were randomly divided into control group,intra-peritoneal injection of 10,30 and 60 mg/kg PA groups(6 in each group),the tumor volume and weight were measured;The expression of Ki-67 and of cleared caspase-3 proteins in tumor tissue was detected by immuno-histochemistry;The expression of Akt/MDM2/p53 signal pathway related protein in tumor tissue was detected by Western blot.Results PA of all doses obviously inhibited proliferation of HT-29 cells,the proportion of cells in S phase and in G_(2)/M phase was deceased and the expression of B cell lymphoma gene-2(Bcl-2),cyclin D1,cycle protein dependent activating enzyme 2(CDK2)proteins all decreased,The ratios of p-Akt/Akt and p-MDM2/MDM2 in a concentration-dependent manner increased(P<0.05).The apoptosis rate,the ratio of cells in G_(0)/G_(1)phase and the protein expression of Bax,cleaved caspase-3 and p53 also increased(P<0.05).PA in all dose groups obviously reduced the tumor volume,weight,the expression of Ki-67 protein and the ratios of p-Akt/Akt,p-MDM2/MDM2 in tumor tissue of nude mice(P<0.05),and increased the protein expression of cleaved caspase-3 and p53 in tumor tissue(P<0.05).Conclusions PA can induce apoptosis and cell cycle arrest of colon cancer cells and inhibit cell proliferation.Its mechanism may be related to the regulation of Akt/MDM2/p53 signal pathway.

关 键 词：结肠癌 茯苓酸 凋亡 细胞周期 Akt/MDM2/p53信号通路 

分 类 号：R735[医药卫生—肿瘤]