首页|β-谷甾醇对结肠癌细胞HCT116增殖和凋亡的影响

β-谷甾醇对结肠癌细胞HCT116增殖和凋亡的影响

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目的 探究β-谷甾醇对结肠癌细胞HCT116增殖和凋亡的影响及其对磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,PKB,又称Akt)信号通路的调控作用.方法 体外培养结肠癌细胞HCT116 并分为β-谷甾醇高(240μmol/L)、中(120μmol/L)、低剂量(60μmol/L)组,设置空白对照组(0μmol/L).应用不同浓度的β-谷甾醇处理HCT116 细胞.24h后应用细胞计数试剂盒 8(cell counting kit-8,CCK-8)法检测β-谷甾醇对HCT116 细胞增殖的影响,并在显微镜下观察细胞的形态变化;应用Hoechst 33342 细胞核染色法检测β-谷甾醇对HCT116 细胞凋亡的影响;应用细胞克隆集落形成实验检测 HCT116 细胞的集落形成能力;应用蛋白质印迹技术检测 HCT116 细胞中PI3K、磷酸化Akt、Akt、抗凋亡蛋白Bcl-2、促凋亡蛋白Bax等相关蛋白的表达情况;利用AutoDock软件实现β-谷甾醇与PI3K和Akt蛋白的分子对接.结果 与对照组比较,β-谷甾醇可浓度依赖性地抑制结肠癌细胞HCT116 的增殖,抑制细胞的集落形成能力,促进细胞的凋亡;抑制HCT116 细胞中磷酸化Akt、PI3K、Bcl-2 蛋白的表达,促进Bax蛋白的表达;β-谷甾醇与PI3K、Akt蛋白的结合均较为稳定.结论 β-谷甾醇可通过抑制PI3K/Akt信号通路调节HCT116 细胞的增殖与凋亡.
Effects of β-sitosterol on proliferation and apoptosis of colon cancer cell HCT116
Objective To investigate the effects of β-sitosterol on the proliferation and apoptosis of colon cancer cell HCT116,and its regulation of on phosphoinositide 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Methods Cultivated colon cancer cells HCT116 in vitro and divided them into β-sitosterol High(240μmol/L)、medium(120μmol/L)and low-dose(60μmol/L)groups,set control group(0μmol/L).Applied different concentrations of β-sitosterol treatment of HCT116 cells.And 24h later,the cell proliferation and activity were determined by cell counting kit-8(CCK-8)method,the morphological changes observed under a microscope;Cell apoptosis was observed by Hoechst 33342 nuclear staining;Used cell colony formation assy to detect the colony forming ability of HCT116 cells;and Western blot was used to evaluate the expression of PI3K,p-Akt,Akt,Bcl-2 and Bax in cells.Used AutoDock software for molecular docking of β-sitosterol with Akt and PI3K.Results Compared with the control group,β-sitosterol could inhibit the proliferation of colon cancer HCT116 cells in a concentration dependent manner,inhibit their colony forming ability and promote cell apoptosis and inhibit the expression of p-Akt、PI3K、and Bcl-2 proteins in HCT116 cells and promotes the expression of Bax protein.The binding of β-sitosterol with PI3K and Akt proteins is relatively stable.Conclusion β-sitosterol may regulate the proliferation and apoptosis of HCT116 through inhibiting PI3K/Akt signaling pathway.

β-sitosterolColon cancerProliferationApoptosisSignaling pathway

陈喜、李若男、李晶、赵李娜、徐志立

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辽宁中医药大学药学院,辽宁大连 116600

β-谷甾醇 结肠癌 增殖 凋亡 信号通路

2024

中国现代医生
中国医学科学院

中国现代医生

影响因子:1.571
ISSN:1673-9701
年,卷(期):2024.62(9)
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