白花蛇舌草乙醇提取物对MHCC97-H、MHCC97-L肝癌细胞体内外增殖的影响  

Effects of Ethanol Extracts of Oldenlandia Diffusa on Proliferation of MHCC97-H and MHCC97-L Hepatocarcinoma Cells In Vitro and In Vivo

作　　者：张宇灵 张树辉[1] ZHANG Yuling;ZHANG Shuhui(Department of Pathology,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院病理科,上海200437

出　　处：《中药药理与临床》2023年第8期77-83,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:81172311);上海中医药大学中西医高原学科项目(编号:30304114325)。

摘　　要：目的:探讨白花蛇舌草乙醇提取物对肝癌细胞MHCC97-H、MHCC97-L增殖的影响。方法:通过CCK-8增殖实验、平板克隆实验、细胞周期及凋亡检测观察白花蛇舌草乙醇提取物对细胞增殖的影响;通过球形成实验、肝癌干细胞标志物EpCAM、CD133检测观察白花蛇舌草乙醇提取物对肝癌干细胞亚群的影响。通过构建MHCC97-H肝癌细胞裸鼠移植瘤模型,观察白花蛇舌草乙醇提取物在体内对肝癌细胞生长的影响,通过构建移植瘤指数生长模型,评估其抗肝癌疗效。结果:白花蛇舌草乙醇提取物呈时间、浓度依赖性抑制肝癌细胞MHCC97-H、MHCC97-L体外增殖。与正常对照组比较,4 mg/mL白花蛇舌草乙醇提取物可降低肝癌细胞克隆形成率(P<0.01),明显增加G_(0)/G_(1)期细胞比例(P<0.01),明显增加凋亡细胞比例(P<0.01),明显降低细胞成球率及EpCAM^(+)、CD133^(+)干细胞亚群比例(P<0.01)。体内实验表明,白花蛇舌草乙醇提取物0.52 g/kg组MHCC97-H移植瘤体积在治疗期内小于生理盐水组,通过构建移植瘤指数增长模型可得生理盐水组与白花蛇舌草乙醇提取物组移植瘤生长速率分别为7.9%,6.8%,白花蛇舌草乙醇提取物0.52 g/kg组肿瘤生长延迟2 d。结论:白花蛇舌草乙醇提取物可能通过阻滞MHCC97-H、MHCC97-L细胞周期于G_(0)/G_(1)期、诱导其凋亡、抑制细胞自我更新能力、抑制EpCAM^(+)、CD133^(+)干细胞亚群生长以抑制肝癌细胞体内外增殖。Objective:To investigate the effects of ethanol extracts of Oldenlandia diffusa on the proliferation of MHCC97-H and MHCC97-L hepatocarcinoma cells in vitro and in vivo.Methods:The effect of ethanol extracts of Oldenlandia diffusa on cell proliferation was detected by CCK-8 assay,plate clone formation assay,and the cell cycle distribution and apoptosis experiments.The effect of ethanol extracts of Oldenlandia diffusa on liver cancer stem cell subsets was determined by sphere-formation assay and detection of liver cancer stem cell markers EpCAM and CD133.The MHCC97-H cell transplanted tumor model in nude mice was established to study the effect of ethanol extracts of Oldenlandia diffusa on the growth of hepatoma cells in vivo,and the tumor exponential growth model was established to explore its anti-cancer effects.Results:The ethanol extracts of Oldenlandia difusa inhibited the proliferation of MHCC97-Hand MHCC97-L cells in vitro in a concentration-and time-dependent manner.Compared to the control group,4 mg/mL ethanol extracts of Oldenlandia difusa significantly decreased the cloning efficiency(P<0.01).The ratio of G_(0)/G_(1)phase cells was increased significantly(P<0.01),and the proportion of apoptotic cells was in creased significantly(P<0.01).The proportion of cell sphericity and EpCAM^(+)/CD133^(+)liver cancer stem cell subsets was decreased significantly(P<0.01).The in vivo experiments showed that the volume of MHCC97-H xenografts in the ethanol extracts of Oldenlandia difusa group was smaller than that in the normal saline group during the treatment period.By constructing the exponential growth model of transplanted tumor,the growth rates of xenografts in the normal saline group and the ethanol extracts of Oldenlandia difusa group were 7.9%and 6.8%,respectively,and the tumor growth was delayed for 2 days in the ethanol extracts of Oldenlandia diffusa group.Conclusion:The ethanol extracts of Oldenlandia difusa may inhibit the proliferation of hepatoma cells in vitro and in vitro by blocking cell cycles of M

关 键 词：白花蛇舌草乙醇提取物 肝细胞癌 增殖 移植瘤指数增长模型 

分 类 号：R285[医药卫生—中药学]