桔梗皂苷D对结直肠癌HT-29细胞增殖、凋亡、迁移和糖酵解的影响  

Effects of platycodon D on proliferation,apoptosis,migration and glycolysis of colorectal cancer HT-29 cell

作　　者：李鹏昊[1] 卜宪越 郝敬鹏[1] Li Penghao;Pu Xianyue;Hao Jingpeng(Department of Anorectal Surgery,the Second Hospital of Tianjin Medical University,Tianjin 300211,China)

机构地区：[1]天津医科大学第二医院肛肠外科,天津300211

出　　处：《国际生物医学工程杂志》2024年第5期423-429,共7页International Journal of Biomedical Engineering

摘　　要：目的探讨桔梗皂苷D对结直肠癌HT-29细胞增殖、凋亡、迁移和糖酵解的影响。方法选取人类HT-29细胞,随机分为对照组及桔梗皂苷D 25、50、100μmol/L组,桔梗皂苷D 25、50、100μmol/L组分别用25、50、100μmol/L桔梗皂苷D处理48 h,通过细胞计数试剂盒-8(CCK-8)和集落形成实验考察细胞增殖情况,采用流式细胞仪和免疫印迹实验考察细胞凋亡和凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和裂解型半胱氨酸天冬氨酸蛋白酶-3(cleaved Caspase-3)的表达,通过Transwell和免疫印迹实验考察细胞迁移情况,并检测葡萄糖消耗、乳酸产生以及HT-29细胞中丙酮酸激酶M2型(PKM2)和磷酸化PKM2(p-PKM2)表达水平。结果随着处理时间的递增,对照组和桔梗皂苷D 25、50、100μmol/L组HT-29细胞存活率逐渐降低;与对照组比较,桔梗皂苷D 25、50、100μmol/L组HT-29细胞存活率均降低(均P<0.001)。与对照组比较,桔梗皂苷D 25、50、100μmol/L组HT-29细胞的集落数由50分别降为36、34、16(均P<0.001)。与对照组相比,桔梗皂苷D 100μmol/L组HT-29细胞的凋亡率由39%提高至62%,Bax相对表达由0.1提高至0.9,cleaved Caspase-3由0.15提高至1.25(均P<0.001),Bcl-2由0.75降低至0.12(P<0.001)。与对照组相比,桔梗皂苷D 100μmol/L组HT-29细胞单位视野数量由48个降低为16个,蜗牛同源转录因子(Snail)相对表达由1.3降低至0.5,上皮钙黏蛋白(E-cadherin)相对表达由0.1提高至1.2,葡萄糖转运蛋白1(GLUT1)的相对表达由1.1降低为0.2,乳酸脱氢酶A(LDHA)的相对表达由1.2降低为0.2,己糖激酶2(HK2)的相对表达由1.00降低为0.15(均P<0.001)。与对照组相比,桔梗皂苷D 100μmol/L组HT-29细胞葡萄糖消耗量由对照组的4.8 mmol/L降低为1.9 mmol/L,乳酸产生量由9.2 mmol/L降低为3.8 mmol/L(均P<0.001)。与对照组相比,桔梗皂苷D 100μmol/L组PKM2的相对表达由1.0降低为0.1,p-PKM2相对水平由0.75降低为0.08(均P<0.001)。结论桔梗皂�Objective To investigate the effects of platycodon D on proliferation,apoptosis,migration and glycolysis of colorectal cancer HT-29 cell.Methods Human HT-29 cells were selected and randomly divided into the control group and platycodon D 25,50,and 100μmol/L groups.The HT-29 cells in the platycodon D 25,50,and 100μmol/L groups were treated with 25,50,and 100μmol/L platycodon D for 48 h.The cell proliferation of the cells was examined by cell counting Kit-8(CCK-8)and colony formation assay.The apoptosis of the cells was investigated using flow cytometry and Western Blot assays were used to examine the expression of apoptosis and apoptosis-related proteins,including B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),and cleaved cystein-asparate protease-3(cleaved Caspase-3).Transwell and Western Blot assays were used to examine cell migration,glucose consumption,and lactic acid production,as well as the expression levels of pyruvate kinase M2(PKM2)and phosphorylated PKM2(p-PKM2)in HT-29 cells.Results With the increase in treatment time,the proliferation rate of HT-29 cells in the control group and platycodon D 25,50,and 100μmol/L groups decreased gradually.Compared with the control group,the viability of HT-29 cells in the platycodon D 25,50,and 100μmol/L groups was decreased(all P<0.001).Compared with the control group,the colony number of HT-29 cells in the platycodon D 25μmol/L(36 vs.50),50μmol/L(34 vs.50),and 100μmol/L(16 vs.50)groups was decreased(all P<0.001),after treatment with platycodon D for 24 h.Compared with the control group,the apoptosis rate of HT-29 cells in the platycodon D 100μmol/L group was increased(62%vs.39%),the relative expression of Bax was increased(0.9 vs.0.1),the cleaved Caspase-3 was increased(1.25 vs.0.15),and the Bcl-2 decreased(0.12 vs.0.75)(all P<0.001).Compared with the control group,the per unit vision number of HT-29 cells in the platycodon D 100μmol/L group decreased(16 vs.48),the relative expression of Snail decreased(0.5 vs.1.3),the relative expression of E-cad

关 键 词：结直肠癌 桔梗皂苷D B淋巴细胞瘤-2 Bcl-2相关X蛋白 裂解型半胱氨酸天冬氨酸蛋白酶-3 丙酮酸激酶M2型 糖酵解 

分 类 号：R285[医药卫生—中药学]