INHBA在胃癌组织中的表达及对胃癌细胞增殖、侵袭转移能力的影响及其机制
Expression of INHBA in gastric cancer tissues and its effect on proliferation, invasion and metastasis of gastric cancer cells
摘要目的:探讨抑制素Beta A(INHBA)在胃癌组织中表达与临床病理资料相关性及对胃癌细胞增殖、侵袭转移能力的影响及机制。方法:利用GEPIA公共数据库验证INHBA在胃癌及癌旁组织中mRNA表达情况及与患者病理分期及预后的关系,利用Kaplan-Meier Plotter在线数据库分析INHBA与患者临床预后的关系;病理标本行免疫组化验证INHBA在胃癌与癌旁组织中蛋白表达水平及其表达水平与患者临床病理分期相关性;体外实验中,利用四唑盐(MTT)比色法检测细胞增殖能力,利用划痕实验检测细胞迁移能力,利用transwell小室实验检测细胞侵袭转移能力,利用蛋白免疫印迹法(Western blot)检测INHBA表达与上皮-间质样转化(EMT)相关蛋白的表达相关性。结果:GEPIA数据库及Kaplan-Meier Plotter在线数据库分析显示,与正常组织相比,INHBA mRNA在多种癌症组织中高表达,在胃癌组织中显著高表达( P<0.05);INHBA mRNA的高表达与胃癌临床分期及不良预后相关( P<0.05);免疫组化结果显示,胃癌组织中INHBA的染色评分显著高于癌旁组织( P<0.05),并且其表达水平与患者临床TNM分期有关( P<0.05);MTT、划痕实验、transwell小室结果显示,INHBA过表达可以促进胃癌细胞增殖、迁移和侵袭转移能力,而干扰INHBA的表达可以抑制细胞增殖、迁移和侵袭转移能力;Western blot结果显示,INHBA过表达后CDH1的表达量明显下调,而CDH2的表达量上调;抑制INHBA表达后CDH1的表达量明显上调,而CDH2的表达量下调。 结论:INHBA在胃癌组织中较癌旁组织高表达,其表达水平与患者肿瘤进展及不良预后相关,INHBA可以促进胃癌MGC-803细胞系的增殖、迁移及侵袭能力,其机制可能与INHBA促进细胞EMT有关。
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abstractsObjective:To investigate the relationship between the expression of inhibin subunit Beta A (INHBA) and the clinicopathological data and its effect on proliferation, invasion and metastasis of gastric cancer cells and its possible mechanisms.Methods:Using Gene Expression Profiling Interactive Analysis (GEPIA) public database to verify the expression of INHBA mRNA in gastric cancer and adjacent tissues and its relationship with pathological stage and prognosis; the relationship between INHBA and clinical prognosis was analyzed using the Kaplan-Meier Plotter online database. Immunohistochemistry was used to confirm the correlation between protein expression level of INHBA in gastric cancer and adjacent tissues and clinical pathological staging. In vitro, the cell proliferation was detected by tetrazolium salt (MTT) method; the cell migration ability was detected by scratch test, and cell invasion and metastasis ability was detected by transwell chamber assay. The expression of INHBA and epithelial-mesenchymal transition (EMT) related proteins was detected by Western blot. Results:The GEPIA database and Kaplan-Meier Plotter online database analysis showed that INHBA mRNA was highly expressed in various cancer tissues and significantly higher in gastric cancer tissues than normal tissues ( P<0.05). The high expression of INHBA mRNA was associated with clinical stage and poor prognosis of gastric cancer ( P<0.05). The results of immunohistochemistry showed that the staining score of INHBA in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.05), and its expression level was correlated with clinical tumor node metastasis (TNM) stage ( P<0.05). The results of MTT, scratch test and transwell chamber showed that INHBA overexpression could promote the proliferation, migration, invasion and metastasis of gastric cancer cells, while interference with INHBA expression could inhibit the proliferation, migration, invasion and metastasis of gastric cancer cells; Western blot results showed that the expression of CDH1 was down regulated and the expression of CDH2 was up-regulated after INHBA overexpression. The expression of CDH1 was up-regulated and the expression of CDH2 was down-regulated after INHBA overexpression was inhibited. Conclusions:INHBA is highly expressed in gastric cancer tissues compared with adjacent tissues. The expression level of INHBA is related to tumor progression and poor prognosis. INHBA can promote the proliferation, migration and invasion of gastric cancer MGC-803 cell line and its mechanism may be related to INHBA promoting cell EMT.
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