miR-199b-3p与CRIM1在胃癌细胞中的靶向关系及其功能  

The targeting relationship and function of miR-199b-3p and CRIM1 in gastric cancer cells

作　　者：王秦西 张炯[1] 车康明 WANG Qinxi;ZHANG Jiong;CHE Kangming(Department of Surgical Oncology,Tianshui First People's Hospital,Tianshui,Gansu 741000,China)

机构地区：[1]甘肃省天水市第一人民医院肿瘤外科,甘肃天水741000

出　　处：《中国普通外科杂志》2024年第10期1679-1687,共9页China Journal of General Surgery

摘　　要：背景与目的:研究发现,miR-199b-3p在胃癌组织中表达下调,而半胱氨酸丰富跨膜BMP调节因子1(CRIM1)在胃癌组织中表达上调。目前miR-199b-3p在胃癌细胞生物学行为中的作用机制尚未明确,其与CRIM1之间是否有关联也不清楚。因此,本研究探讨是否miR-199b-3p与CRIM1存在相互作用以及对胃癌细胞功能的影响。方法:采用qRT-PCR与免疫组化方法检测胃癌组织与癌旁组织中miR-199b-3p和CRIM1的表达。以胃癌MGC803细胞为研究对象,观察过表达miR-199b-3p(miR-199b-3p模拟物)与敲低CRIM1(si-CRIM1)后,细胞增殖能力、侵袭/迁移能力以及凋亡率的变化;应用生物信息学方法预测和双荧光素酶报告基因实验分析miR-199b-3p和CRIM1的靶向关系,并用Western blot方法验证。结果:qRT-PCR结果显示,与癌旁组织比较,胃癌组织中miR-199b-3p表达水平降低,而CRIM1增加(均P<0.05);免疫组化实验结果显示,CRIM1在癌组织中呈阳性表达,在癌旁组织呈阴性表达。过表达miR-199b-3p或敲低CRIM1后,MGC803细胞的增殖与侵袭/迁移能力降低,凋亡率增加(均P<0.05)。生物信息学方法预测和双荧光素酶报告基因实验显示miR-199b-3p与CRIM1存在靶向关系;Western blot实验结果表明,转染miR-199b-3p模拟物后,CRIM1表达降低(P<0.05)。结论:CRIM1是miR-199b-3p的靶基因,miR-199b-3p能够通过靶向调控CRIM1的活性进而抑制胃癌细胞的增殖、侵袭、迁移,并促进胃癌细胞凋亡,miR-199b-3p/CRIM1通路可作为胃癌治疗的潜在靶点进一步研究。Background and Aims:Studies have shown that miR-199b-3p is downregulated in gastric cancer tissues,while cysteine-rich transmembrane BMP regulator 1(CRIM1)is upregulated in these tissues.However,the role and mechanism of miR-199b-3p in the biological behavior of gastric cancer cells are still unclear,as is its potential association with CRIM1.Therefore,this study was conducted to investigate whether there is an interaction between miR-199b-3p and CRIM1 and how they affect the function of gastric cancer cells.Methods:qRT-PCR and immunohistochemistry were used to detect the expression levels of miR-199b-3p and CRIM1 in gastric cancer tissues and adjacent non-cancerous tissues.Gastric cancer MGC803 cells were used to assess changes in cell proliferation,invasion/migration abilities,and apoptosis rates after overexpression of miR-199b-3p(using miR-199b-3p mimics)or knockdown of CRIM1(using siCRIM1).Bioinformatics analysis was used to predict the targeting relationship between miR-199b-3p and CRIM1,which was further validated by dual-luciferase reporter assay and confirmed through Western blot analysis.Results:The results of qRT-PCR indicated that,compared to adjacent non-cancerous tissues,miR-199b-3p expression was significantly lower in gastric cancer tissues,while CRIM1 expression was higher(both P<0.05).Immunohistochemistry results demonstrated positive expression of CRIM1 in cancerous tissues,while it was negative in non-cancerous tissues.Overexpression of miR-199b-3p or CRIM1 knockdown resulted in decreased proliferation and invasion/migration abilities of MGC803 cells,along with increased apoptosis rates(all P<0.05).Bioinformatics prediction and dual-luciferase reporter assays confirmed that CRIM1 is a target of miR-199b-3p.Western blot analysis showed that CRIM1 expression was significantly reduced after transfection with miR-199b-3p mimics(P<0.05).Conclusion:CRIM1 is a target gene of miR-199b-3p,which can inhibit the proliferation,invasion,and migration of gastric cancer cells while promoting apoptosis by tar

关 键 词：胃肿瘤 微RNAS CRIM1 细胞增殖 肿瘤侵润 

分 类 号：R735.2[医药卫生—肿瘤]