摘要
目的探究抑制剂PD98059阻断MAPK/ERK信号通路对结直肠癌细胞HT29增殖抑制和凋亡促进的作用。方法体外培养人正常结肠黏膜上皮细胞NCM460和结直肠癌细胞HT29,检测细胞中ERK mRNA及蛋白的表达;分别以25 mmol/L、50 mmol/L、100 mmol/L、200 mmol/L、300 mmol/L及400 mmol/L的PD98059处理HT29细胞,并设立对照组和空白组,检测HT29细胞增殖、周期和凋亡情况及相关蛋白的变化。结果与NCM460细胞相比,HT29细胞中ERK mRNA和蛋白表达水平均上升(均P <0.05);添加抑制剂PD98059后,HT29细胞增殖活性降低(P <0.05),且浓度为0~100 mmol/L时,抑制效率较高;与对照组相比,添加抑制剂PD98059后,HT29细胞生长停滞于G0/G1期,凋亡率、p21和caspase-3的表达水平均升高(均P <0.05),ERK mRNA和蛋白、cycling D1的表达水平均降低(均P <0.05),具有剂量依赖性。结论结直肠癌细胞中MAPK/ERK信号通路被激活,抑制剂PD98059可阻断MAPK/ERK通路抑制HT29细胞的增殖,促进其凋亡,可能与调控cycling D1、p21及caspase-3表达水平有关。
Objectives To investigate the effect of inhibitor PD98059 blocking MAPK/ERK pathway on inhibiting proliferation and promoting apoptosis of colorectal cancer cells. Methods Human normal colonic mucosal epithelial cells NCM460 and colorectal cancer cells HT29 were cultured in vitro. The expression of ERK was detected. HT29 cells were treated with 25, 50,100, 200, 300 and 400 mmol/L PD98059, respectively. And control group and blank group were set up. Changes in proliferation, cycle, apoptosis, and related proteins were detected. Results Compared with NCM460 cells, expression of ERK mRNA and protein significantly increased in HT29 cells(P < 0.05). The proliferation of HT29 cells significantly decreased with inhibitor PD98059(P < 0.05), with the higher inhibition effect when concentration is of 0~100 mmol/L. Compared with the control, HT29 cells growth was blocked in G0/G1 phase with inhibitor PD98059, and the apoptosis rate, the expression of p21 and caspase-3 significantly increased(P < 0.05), while the expression of ERK mRNA and protein, and cycling D1 significantly decreased(P <0.05) in a dose-dependence manner. Conclusion MAPK/ERK signaling pathway is activated in colorectal cancer cells. The inhibitor PD98059 can block MAPK/ERK pathway, inhibit proliferation of HT29 cells, and promote apoptosis, which may be related to regulating cycling D1, p21 and caspase-3 levels.
作者
杨扬
夏冬琴
王维
Yang Yang;Xia Dongqin;Wang Wei(Department of Traditional Chinese Medicine and Oncology,Chongqing University Cancer Hospital,Chongqing 400030,China)
出处
《结直肠肛门外科》
2019年第6期657-661,667,共6页
Journal of Colorectal & Anal Surgery
基金
重庆市卫生和计划生育委员会科研课题(20170300182)
