摘要
目的探讨Wnt3a对结肠癌SW480细胞增殖的调控效应及其作用机制。方法将培养好的结肠癌细胞SW480随机分为对照组和Wnt3a组,对照组仅给予DMEM培养基处理,Wnt3a组给予100 ng/m L Wnt3a培养。用MTT法检测细胞增殖情况,计算相对活细胞百分比;用平板克隆形成实验检测细胞克隆形成率(PE);用流式细胞术检测细胞周期分布;用Western blotting法检测细胞内醛脱氢酶1B1(ALDH1B1)蛋白表达,RT-PCR技术检测ALDH1B1 mRNA表达。结果 Wnt3a诱导24、48、72 h,Wnt3a组相对活细胞百分比高于对照组(P均<0.05)。Wnt3a诱导7 d,Wnt3a组PE高于对照组(P<0.05)。Wnt3a诱导24 h,Wnt3a组G1期细胞百分比低于对照组,G2期细胞百分比高于对照组(P均<0.05)。Wnt3a诱导48 h,Wnt3a组ALDH1B1蛋白、mRNA相对表达量高于对照组(P均<0.05)。结论 Wnt3a可能通过诱导ALDH1B1高表达促进结肠癌SW480细胞增殖。
Objective To investigate the potential role of Wnt3 a in the regulation of colon cancer cell proliferation and its regulatory mechanism. Methods Colon carcinoma cell line SW480 was randomly divided into two groups,the control group treated with DMEM,and the Wnt3 a group treated with 100 ng / m L Wnt3 a. The proliferation ability was measured by MTT. The relative percentage of living cells was also counted. Cell clone formation rate（ PE） was measured by plate clone formation assay. Cell cycle was analyzed by flow cytometry. RT-PCR was used to measure the expression level of ALDH1B1 mRNA. The expression level of aldehyde dehydrogenase（ ALDH） 1B1 protein was measured by Western blotting.Results After 24,48 and 72 h of induction by Wnt3 a,the relative percentage of living cells in the Wnt3 a group was significantly higher than that of the control group（ all P〈0. 05）. After 7 d of induction by Wnt3 a,the PE of the Wnt3 a group was significantly higher than that of the control group（ P〈0. 05）. The cell percentage of G1 phase in the Wnt3 a group was significantly lower,and the cell percentage of G2 phase was significantly higher than that of the control group after 24-hour treatment of Wnt3a（ all P〈0. 05）. After 48-hour treatment with Wnt3 a,the expression level of ALDH1B1 protein and mRNA in the Wnt3 a group was significantly higher than that of the control group（ all P〈0. 05）. Conclusion Wnt3 a promotes colon cancer cell proliferation by inducing the high expression of ALDH1B1.
作者
马松林
张姮
廖宇圣
徐丹
吴杰
MA Songlin ZHANG Heng LIAO Yusheng XU Dan WU Jie(Wuhan Central Hospital Affiliated to Huazhong University of Science and Technology, Wuhan 430014, China)
出处
《山东医药》
CAS
北大核心
2016年第37期18-20,共3页
Shandong Medical Journal
