摘要
目的观察甘精胰岛素在体外对结肠癌LoVo细胞的影响并探讨其可能机制。方法使用不同浓度甘精胰岛素对结肠癌LoVo细胞进行干预,以人胰岛素进行对照,分别通过CCK8法和流式细胞术检测结肠癌细胞的增殖和凋亡,通过蛋白印迹法检测ERK1/2蛋白表达。结果经甘精胰岛素(20、50、100、150、200 n M)处理后LoVo细胞数量明显增加,与空白组比较,甘精胰岛素(100、150、200 n M)和阳性对照组人胰岛素(100 n M)有显著促增殖作用(P<0.05)。空白组早期细胞凋亡率为(13.26±1.34)%,甘精胰岛素(100、150、200 n M)处理后凋亡率分别减少到(11.52±2.18)%、(9.11±1.39)%和(6.08±1.92)%。加入ERK1/2抑制剂PD98059后甘精胰岛素未显示出诱导LoVo细胞增殖和减少凋亡的作用。甘精胰岛素上调ERK1/2磷酸化水平并呈时间和剂量依赖性,但ERK1/2总水平没有改变。结论甘精胰岛素在体外有诱导结肠癌LoVo细胞增殖的作用,其机制可能是通过激活ERK1/2信号转导通路实现的。
Objective To observe the roles of glargine on colon cancer LoVo cells in vitro and to investigate the potential mechanisms. Methods Colon cancer LoVo cells were treated with different concentrations of glargine and human insulin. Proliferation of the cells treated with glargine was assayed by CCK8. Apoptosis in the treated cells was measured by flow cytometry. The protein expression of ERK1 /2 in treated cells was determined by Western blot.Results Glargine( 20,50,100,150,200 n M) dramatically increased the LoVo cells number. Compared with the control,both glargine( 100,150,200 n M) and human insulin( 100 n M) as the positive control induced significant cells proliferation( P < 0. 05). The rate of cell early apoptosis in the control was 13. 26%,while the rate decreased to11. 52%,9. 11% and 6. 08% after the cells treated with glargine at 100,150 and 200 n M,respectively. After ERK1 /2 inhibitor PD98059 was added into LoVo cells,proliferation was not induced by glargine and early apoptosis rate was not innduced. An increase in glargine( 200 n M) stimulated p-ERK1 /2 occurred in a time and dose dependent manner. However,the total protein levels of ERK1 /2 remained unaffected throughout the course of these experiments. Conclusion Glargine can induce the proliferation of colon cancer LoVo cells in vitro. ERK1 /2 signal pathway activation may be the potential mechanism.
出处
《中国临床新医学》
2015年第10期897-901,共5页
CHINESE JOURNAL OF NEW CLINICAL MEDICINE
基金
国家自然科学基金资助项目(编号:81160107)
广西科学研究与技术开发项目(编号:桂科攻1140003B-69)
广西自然科学基金资助项目(编号:2010GXNSFB013083)
