학술논문
MEX3A对结肠癌细胞恶性表型的影响及其机制研究 / Effects of MEX3A on malignant phenotype of colon cancer cells and its mechanism of action
Document Type
Academic Journal
Author
周俊邑; 沙晓锋; 柳叶; 韩卓莹; 丁姝; 黄豪; 罗超; 仲小敏; ZHOU Junyi; SHA Xiaofeng; LIU Ye; HAN Zhuoying; DING Shu; HUANG Hao; LUO Chao; ZHONG Xiaomin
Source
现代肿瘤医学 / Journal of Modern Oncology. 32(2):226-233
Subject
Language
Chinese
ISSN
1672-4992
Abstract
目的:探讨MEX3A在结肠癌组织中的表达及其对结肠癌细胞恶性表型的影响并探索其机制.方法:TCGA数据库分析MEX3A在结肠癌组织及正常组织中的差异表达及其与患者临床病理分期和预后的相关性,并通过免疫组化进一步验证.采用慢病毒对结肠癌细胞中MEX3A的表达进行敲低,检测细胞的增殖、凋亡、迁移及侵袭能力.裸鼠皮下荷瘤后对肿瘤生长进行测量记录.Flag融合表达质粒过表达MEX3A后进行COIP实验.采用质谱进行蛋白质定性检测(Shotgun)与MEX3A互作的蛋白质.生物信息学分析后进行COIP验证,对互作蛋白表达进行挽救实验验证.结果:MEX3A在结肠癌组织中表达显著上调且与患者临床病理特征及预后相关.采用慢病毒敲减MEX3A表达后,结肠癌细胞增殖能力显著被抑制,凋亡则显著增加.细胞转移能力检测结果显示,结肠癌细胞的迁移及侵袭能力显著被抑制.小鼠体内实验结果表明,敲减MEX3A表达后,肿瘤在体内的生长明显被抑制.蛋白质定性检测及COIP验证结果显示,EIF2AK2、LAMB3及MSI2与MEX3A蛋白存在相互作用.在敲减MEX3A表达后分别过表达EIF2AK2、LAMB3及MSI2后细胞增殖能力被不同程度恢复,其中MSI2过表达后细胞增殖能力恢复最明显.MTT及细胞迁移能力验证结果显示过表达MSI2后,细胞增殖及迁移能力显著恢复.结论:MEX3A在结肠癌组织中表达上调且与患者预后呈负相关,其通过与MSI2相互作用促进结肠癌细胞的恶性表型.
Objective:To investigate the expression of MEX3A in colon cancer tissue and its effect on the malig-nant phenotype of colon cancer cells and explore its mechanism.Methods:The TCGA database analyzed the differen-tial expression of MEX3A in colon cancer tissues and normal tissues and its correlation with the clinicopathological stage and prognosis of patients,and the results were further verified by immunohistochemistry.Lentiviral was used to knock down the expression of MEX3A in colon cancer cells to detect the proliferation,apoptosis,migration and inva-sion ability of cells.Tumor growth was measured and recorded after subcutaneous tumors in nude mice.COIP experi-ments were performed after Flag fusion expression plasmid overexpression MEX3A.Qualitative detection of proteins was performed using mass spectrometry(Shotgun)interacting with MEX3A.After bioinformatics analysis,COIP veri-fication was performed,and rescue experiments were performed to verify the expression of interactive proteins.Re-sults:The expression of MEX3A in colon cancer tissue was significantly upregulated and correlated with the clinico-pathological features and prognosis.After lentiviral knockdown MEX3A expression,the proliferation ability of colon cancer cells was significantly inhibited,and apoptosis was significantly increased.The results of cell metastasis ability test showed that the migration and invasion ability of colon cancer cells were significantly inhibited.The results of in vivo experiments in mice showed that the growth of tumors in vivo was significantly inhibited after knocking out MEX3A expression.Qualitative protein detection and COIP verification results showed that EIF2AK2,LAMB3 and MSI2 interacted with MEX3A proteins.After the expression of knock-down MEX3A,the cell proliferation ability was restored to different degrees after overexpression of EIF2AK2,LAMB3 and MSI2,among which the cell proliferation a-bility was the most obvious after MSI2 overexpression.The results of MTT and cell migration ability verification showed that after overexpression of MSI2,cell proliferation and migration ability were significantly restored.Conclu-sion:The expression of MEX3A in colon cancer tissue is upregulated and the prognosis of patients is negatively corre-lated with MEX3A high expression,and it promotes the malignant phenotype of colon cancer cells by interacting with MSI2.
Objective:To investigate the expression of MEX3A in colon cancer tissue and its effect on the malig-nant phenotype of colon cancer cells and explore its mechanism.Methods:The TCGA database analyzed the differen-tial expression of MEX3A in colon cancer tissues and normal tissues and its correlation with the clinicopathological stage and prognosis of patients,and the results were further verified by immunohistochemistry.Lentiviral was used to knock down the expression of MEX3A in colon cancer cells to detect the proliferation,apoptosis,migration and inva-sion ability of cells.Tumor growth was measured and recorded after subcutaneous tumors in nude mice.COIP experi-ments were performed after Flag fusion expression plasmid overexpression MEX3A.Qualitative detection of proteins was performed using mass spectrometry(Shotgun)interacting with MEX3A.After bioinformatics analysis,COIP veri-fication was performed,and rescue experiments were performed to verify the expression of interactive proteins.Re-sults:The expression of MEX3A in colon cancer tissue was significantly upregulated and correlated with the clinico-pathological features and prognosis.After lentiviral knockdown MEX3A expression,the proliferation ability of colon cancer cells was significantly inhibited,and apoptosis was significantly increased.The results of cell metastasis ability test showed that the migration and invasion ability of colon cancer cells were significantly inhibited.The results of in vivo experiments in mice showed that the growth of tumors in vivo was significantly inhibited after knocking out MEX3A expression.Qualitative protein detection and COIP verification results showed that EIF2AK2,LAMB3 and MSI2 interacted with MEX3A proteins.After the expression of knock-down MEX3A,the cell proliferation ability was restored to different degrees after overexpression of EIF2AK2,LAMB3 and MSI2,among which the cell proliferation a-bility was the most obvious after MSI2 overexpression.The results of MTT and cell migration ability verification showed that after overexpression of MSI2,cell proliferation and migration ability were significantly restored.Conclu-sion:The expression of MEX3A in colon cancer tissue is upregulated and the prognosis of patients is negatively corre-lated with MEX3A high expression,and it promotes the malignant phenotype of colon cancer cells by interacting with MSI2.