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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 of glutathione peroxidase 1 (GPX1) in colorectal cancer (CRC) and its effect on the proliferation, migration and invasion of CRC cell lines HT29 and LOVO Impact. Methods: From June 2015 to March 2016, 60 cases of clinical CRC tissues and adjacent non-cancerous tissues resected by general surgery in Binjiang Branch of Haikou People’s Hospital were collected. Real-time quantitative PCR (qPCR) GPX1 m RNA levels in tissues. The GPX1-knockout cell line was constructed by lentivirus-carrying sh RNA infection in HT29 cell line overexpressing GPX1 m RNA. GPX1-overexpressing cell lines were constructed by transient transfection in the LOVO cell line expressing low GPX1 m RNA GPX1 m RNA and protein levels were verified. MTS assay was used to detect the changes of CRC cell proliferation. The changes of invasion and migration of CRC cells were detected by scratch assay and Transwell invasion assay respectively. The expressions of E-cadherin and vimentin were detected by Western blotting. Results: The expression of GPX1 m RNA in CRC tissues was significantly lower than that in adjacent noncancerous tissues (0.051 ± 0.024 vs 0.142 ± 0.051, P <0.01). After GPX1 knockdown, the proliferation of HT29 cells was significantly enhanced (12.901 ± 2.790 vs 6.617 ± 2.462, P <0.01), and the invasion ability was significantly enhanced [(384.7 ± 37.9 vs 209.2 ± 31.2, P <0.01] (0.139 ± 0.025) vs (0.251 ± 0.038) mm, P <0.01]. The expression of E-cadherin was down-regulated (P <0.01) and the expression of vimentin was up-regulated (P <0.05). The proliferation of LOVO cells overexpressing GPX1 was significantly decreased (P <0.01), migration and invasion were decreased (P <0.05), E-cadherin was upregulated (P <0.01) . CONCLUSION: GPX1 m RNA is low expressed in human CRC tissues. GPX1 negatively regulates the proliferation, migration and invasion of CRC cells, which may play a role in tumor suppressor in CRC.