目的:观察过表达MAWBP对结直肠癌细胞增殖、侵袭、迁移的影响及其相关分子机制。方法:将caco-2、HT-29细胞分为过表达MAWBP组和空载组,用慢病毒载体构建稳定过表达MAWBP的结直肠癌细胞株,Western blot和q-PCR实验验证细胞中MAWBP的表达水平;MTT实验检测细胞的增殖能力;Transwell实验检测细胞的侵袭和迁移能力;Western blot实验检测细胞中EMT以及MAPK通路标志蛋白的表达情况。结果:过表达MAWBP组中MAWBP的蛋白和mRNA的表达水平较空载组升高;MTT和Transwell实验分别提示过表达组细胞的增殖和迁移侵袭率较空载组降低(P〈0.05);过表达组细胞中EMT标志蛋白E-cadherin表达上调,Snail表达下调,MAPK通路标志蛋白p-ERK和p-38表达水平下调。结论:过表达MAWBP可抑制结直肠癌细胞的增殖、侵袭和迁移的能力,这种抑制作用可能是通过抑制MAPK通路和调控上皮细胞间充质转化过程而产生。
Objective To study the effects and molecular mechanism of MAWBP in the proliferation,invasion and migration of human colorectal cancer cell lines. Methods Colorectal cancer cell lines, caco-2 and HT-29, were divided into MAWBP over-expressed group and control group. MAWBP was stablely over-expressed in both cell lines by lentivirus, and the positive clones were screened by puromycin. The expression of MAWBP gene at m RNA and protein levels was evaluated by RT-PCR and western blot. Cell proliferation was detected by MTT. Cell migration and invasion were detected by transwell experiment. The EMT related protein levels of Snail,E-cadherin, and MAPK pathway protein of ERK, p38, p-ERK, p-p38 were determined by western blot analysis.Results Compared with the control group, the expression of MAWBP was significantly higher in HT-29-MAWBP and caco2-MAWBP group. The cell proliferation, invasion and migration ability significantly reduced in the HT-29-MAWBP group. The expression of Snail, p-ERK and p-p38 significantly decreased, while the Ecadherin increased in HT-29-MAWBP and caco2-MAWBP group. Conclusions Over- expression of MAWBP exerts an inhibitory effect on the proliferation, invasion and migration of colorectal caner cells via inhibit the MAPK pathway and regulate the EMT process.