β-谷甾醇对结肠癌细胞HCT116增殖和凋亡的影响

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.