加减半夏泻心汤对胃癌细胞HCG-27细胞凋亡及其生物学行为的影响和机制初探  

Effect and mechanism of banxia Xiexin Decoction on apoptosis and biological behavior of HCG-27 cells

作　　者：王震凯 陈丽丽 陈婉珍 刘翔[1] 刘丹 杨晓倩 周冬梅 姜志欣 WANG Zhenkai;CHEN Lili;CHEN Wanzhen;LIU Xiang;LIU Dan;YANG Xiaoqian;ZHOU Dongmei;JIANG Zhixin(Endoscopy Center,Nanjing Hospital of Traditional Chinese Medicine,Nanjing,Jiangsu 210008,China;Health Center of Pengjie Town,Luqiao District,Taizhou,Zhejiang 318057,China;Jiangsu Health Development Research Center,Nanjing,Jiangsu 210010,China)

机构地区：[1]南京市中医院内镜中心,江苏南京210008 [2]浙江省台州市路桥区蓬街镇卫生院,浙江台州318057 [3]江苏省卫生健康发展研究中心,江苏南京210010

出　　处：《医药前沿》2023年第6期1-6,共6页Journal of Frontiers of Medicine

基　　金：加减半夏泻心汤治疗胃癌前病变的临床观察及干预机制研究,2021年度江苏省开放课题(JSHD2021025)。

摘　　要：目的:研究加减半夏泻心汤药物血清对胃癌细胞HCG-27细胞凋亡和Wnt信号通路中Wnt1和β-catenin蛋白表达的影响。方法:采用噻唑蓝(MTT)比色法检测不同浓度(5%、10%和20%)加减半夏泻心汤药物血清对HCG-27细胞增殖能力的影响;利用Annexin V-FITC和流式细胞术检测其对细胞凋亡的影响;利用软琼脂集落形成实验检测其对细胞非锚定依赖性生长能力的影响;采用蛋白免疫印迹法(Western-blot)和qRT-PCR检测其对Wnt信号通路中Wnt1和β-catenin蛋白表达的影响。结果:MTT结果显示,加减半夏泻心汤药物血清能够抑制HCG-27细胞的增殖能力,随药物血清浓度升高而增强;Annexin V-FITC和流式细胞术显示,经过10%和20%浓度的药物血清处理后的HCG-27细胞凋亡率显著高于空白对照组,差异有统计学意义(P<0.05);软琼脂集落形成实验提示,10%和20%浓度的药物血清均能显著抑制胃癌细胞非锚定依赖性生长能力,且随着药物浓度的增加而增高,差异有统计学意义(P<0.05);10%和20%浓度的药物血清的Wnt1和β-catenin蛋白表达均下降,差异有统计学意义(P<0.05)。结论:加减半夏泻心汤能够抑制HCG-27细胞增殖和非锚定依赖性生长能力,诱导细胞凋亡,其作用机制可能是通过抑制经典Wnt信号通路进行调控。Objective To investigate the effects of banxia Xiexin Decoction on the apoptosis of HCG-27 cells and the expression of Wnt1 andβ-catenin protein in Wnt signaling pathway.Methods MTT(thiazolyl blue)colorimetric assay was used to detect the effects of banxia Xiexin Decoction serum(5%,10%and 20%)on the proliferation of HCG-27 cells.AnnexinⅤ-FITC and flow cytometry were used to determine the effect of AnnexinⅤ-FITC on apoptosis.Soft AGAR colony formation assay was used to detect the effect of colony formation on unanchored cell growth.The expression of Wnt1 andβ-catenin in Wnt signaling pathway was detected by Western-blot and QRT-PCR.Results MTT results showed that adding or reducing banxia Xiexin Decoction serum could inhibit the proliferation of HCG-27 cells,which increased with the increase of drug serum concentration.AnnexinⅤ-FITC and flow cytometry showed increased apoptosis rate of HCG-27 cells treated with 10%and 20%banxia Xiexin Decoction serum compared with blank control group(P<0.05 and P<0.001,respectively).The soft AGAR colony formation experiment showed that both 10%and 20%banxia Xiexin Decoction serum could significantly inhibit the non-anchoring growth of gastric cancer cells,which increased with the increase of banxia Xiexin Decoction serum concentration(P<0.05 and P<0.001,respectively).The expression of Wnt1 andβ-catenin decreased(P<0.05 and P<0.001,respectively).Conclusion Banxia Xiexin Decoction can inhibit the proliferation and non-anchoring growth of HCG-27 cells and induce apoptosis by inhibiting classical Wnt signaling pathway.

关 键 词：半夏泻心汤 胃癌 细胞凋亡 WNT信号通路 

分 类 号：R735.2[医药卫生—肿瘤]