摘要
目的:探讨谷胱甘肽过氧化物酶1(glutathione peroxidase 1,GPX1)在结直肠癌(colorectal cancer,CRC)组织中的表达水平及其对CRC细胞株HT29和LOVO细胞增殖、迁移及侵袭能力的影响。方法:收集2015年6月至2016年3月间海口市人民医院滨江分院普外科手术切除的60例临床CRC组织及癌旁组织,应用实时荧光定量PCR(q PCR)技术检测CRC组织及癌旁组织中GPX1 m RNA水平。在高表达GPX1 m RNA的HT29细胞株用慢病毒携带sh RNA感染的方法构建敲除GPX1的细胞株,瞬时转染法在低表达GPX1 m RNA的LOVO细胞系构建过表达GPX1的细胞株,并在GPX1 m RNA及蛋白水平进行验证。通过MTS法检测CRC细胞的增殖能力的变化,用划痕实验、Transwell侵袭实验分别检测CRC细胞迁移侵袭能力的变化,同时用Western blotting检测E-钙黏蛋白和波形蛋白表达的变化。结果:CRC组织中GPX1 m RNA表达水平显著低于癌旁组织(0.051±0.024 vs 0.142±0.051,P<0.01)。敲除GPX1后,HT29细胞的增殖能力显著增强(12.901±2.790 vs 6.617±2.462,P<0.01)、侵袭能力显著增强[(384.7±37.9)vs(209.2±31.2)个,P<0.01]、迁移能力显著增强[(0.139±0.025)vs(0.251±0.038)mm,P<0.01]、E-钙黏蛋白表达下调(P<0.01)、波形蛋白表达上调(P<0.05)。过表达GPX1的LOVO细胞的增殖能力显著降低(P<0.01)、迁移和侵袭能力下降(均P<0.05)、E-钙黏蛋白上调(P<0.01)、波形蛋白表达下调(P<0.01)。结论:GPX1 m RNA在人CRC组织中低表达,GPX1负向调控CRC细胞的增殖、迁移和侵袭,其在CRC中可能发挥抑癌作用。
Objective：To investigate the expression level of glutathione peroxidase 1（GPX1）in colorectal cancer（CRC） tissues,and its effect on the proliferation and migration of CRC HT29 and LOVO cell lines.Methods：Colorectal cancer tissues and para-cancerous tissues that resected from 60 CRC patients at Binjiang Branch of Haikou People＇s Hospital from June,2015 to March,2016 were collected for this study.Real-time fluorescence quantitative PCR was used to detect the expression level of GPX1 m RNA in sample tissues.HT29 cell line expressing higher GPX1 m RNA was transfected with lentivirus sh RNA to stably knock-down GPX1 expression,while LOVO cell line expressing lower GPX1 m RNA was subjected to transient transfection method to stably overexpressing GPX1,and the protein and m RNA expression level of GPX1 were verified in cells.The change in cell proliferation ability was detected by MTS assay,and the invasion and migration ability was detected by Transwell invasion assay and wound healing assay.The changes in expression of E-cadherin and vimentin were detected by Western blotting.Results：GPX1 m RNA expression was significantly lower in CRC tissues compared with para-cancerous tissues（0.051±0.044 vs 0.142±0.051,P〈0.01）.After knock-down of GPX1：the proliferation ability of HT29 cells was significantly improved（12.901±2.790 vs 6.617±2.462,P〈0.01）,the invasion and migration ability of HT29 cells were significantly enhanced（invasion：[384.7±37.9] vs [209.2± 31.2],P〈0.01;migration：[0.139±0.025] mm vs [0.251±0.038] mm,P〈0.01）;and there was a down-regulation of E-cadherin（P〈0.01） and an up-regulation of vimentin（P〈0.01）.After overexpression of GPX1：the proliferation ability of LOVO cell was significantly decreased（P〈0.01）;the invasion and migration ability of LOVO cell was significantly decreased（all P〈0.01）;and there was an up-regulation of E-cadherin（P〈0.01） and a down-regulation of vimentin（P〈0.01）.Conclusion：GPX1 m RNA was low-expressed in human colorectal cancer tissues.GPX1 may inhibit the proliferation,invasion and migration of CRC cell lines,and may play an important anti-carcinoma role in CRC.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2017年第9期995-1001,共7页
Chinese Journal of Cancer Biotherapy
关键词
谷胱甘肽过氧化物酶-1
结直肠癌
增殖
侵袭
迁移
glutathione peroxidase 1（GPX1）
colorectal cancer
proliferation
invasion
metastasis
