麦冬皂苷B对结肠癌细胞增殖、凋亡的影响及其机制研究

Effect of Ophiopogonin B on Proliferation and Apoptosis of Colon Cancer Cells and Its Mechanism

目的探讨麦冬皂苷B对结肠癌细胞增殖、凋亡的影响及其机制。方法选取本院2017年1月~2019年1月收治的25例结肠癌患者的癌组织及相应癌旁组织;将结肠癌细胞HT29分为对照(NC)组、不同浓度麦冬皂苷B组、miR-NC组、miR-142-3p组、anti-miR-NC组、anti-miR-142-3p组、麦冬皂苷B+anti-miR-NC组、麦冬皂苷B+anti-miR-142-3p组。细胞计数试剂盒8(CCK-8)检测细胞增殖;蛋白质印迹(Western blot)法检测细胞周期蛋白D1(cyclin D1)、裂解的半胱氨酸天冬氨酸蛋白酶3(cleaved-caspase-3)蛋白表达;流式细胞术检测细胞凋亡;实时荧光定量PCR(RT-qPCR)检测miR-142-3p表达水平。结果麦冬皂苷B处理的结肠癌细胞HT29中cyclin D1表达水平显著降低,cleaved-caspase-3表达水平显著升高,细胞凋亡率显著升高,miR-142-3p表达水平显著升高(P<0.05)。与癌旁组织相比,结肠癌组织中miR-142-3p表达水平显著降低(P<0.05);mi R-142-3p高表达可抑制结肠癌细胞增殖、促进细胞凋亡;mi R-142-3p低表达可促进结肠癌细胞增殖、抑制细胞凋亡;mi R-142-3p低表达可抑制麦冬皂苷B对结肠癌细胞HT29增殖和凋亡的影响。结论麦冬皂苷B通过上调miR-142-3p的表达抑制结肠癌细胞增殖、促进细胞凋亡。

Objective To investigate the effects and mechanism of saponin B on proliferation and apoptosis of colon cancer cells. Methods The cancerous tissues and corresponding adjacent tissues of 25 patients with colon cancer treated in our hospital from January 2017 to January 2019 were collected. The colon cancer cells(HT29) were divided into control(NC) group, groups treated with different concentrations of ophiopogonin B, miR-NC group, miR-142-3 p group, anti-miR-NC group, anti-miR-142-3 p group, ophiopogonin B+anti-miR-NC group and ophiopogonin B+antimiR-142-3 p group. The cell proliferation was detected by cell counting kit 8(CCK-8);expression levels of cyclin D1 and cleaved cysteine aspartic protease-3(cleaved-caspase-3) were detected by Western blot;the cell apoptosis was detected by flow cytometry;the expression level of miR-142-3 p was detected by real-time fluorescent quantitative PCR(RT-qPCR). Results The expression level of cyclin D1 in the colon cancer cells(HT29) was significantly reduced, the expression level of cleaved-caspase-3 significantly increased, the apoptosis rate significantly increased, and the expression level of miR-142-3 p significantly increased(P<0.05). Compared with adjacent tissues, the expression of miR-142-3 p in colon cancer tissue was significantly reduced(P<0.05). Overexpression of miR-142-3 p could inhibit proliferation of colon cancer cells and promote apoptosis. Low expression of miR-142-3 p could promote proliferation of colon cancer cells and inhibit apoptosis. Low expression of miR-142-3 p could inhibit the effect of ophiopogonin B on the proliferation and apoptosis of colon cancer cells HT29. Conclusion Ophiopogonin B can inhibit proliferation of colon cancer cells and promote apoptosis by up-regulating the expression of miR-142-3 p.