| 摘要: |
| ［摘要］ 目的 探讨miR-145-5p在结直肠癌组织和结直肠癌细胞中的表达及其通过GTP结合蛋白2（GTPBP2）调节结直肠癌的机制。方法 采用实时荧光定量PCR（qPCR）法检测44例结直肠癌组织及对应癌旁组织、结直肠癌细胞（HCT116）及人正常结直肠黏膜细胞(FHC）中miR-145-5p和GTPBP2 mRNA的表达水平。采用双荧光素酶报告基因实验验证miR-145-5p与GTPBP2基因的靶向关系。将miR-145-5p模拟物（miR-145-5p mimic）、miR-145-5p模拟物阴性对照（mimic-NC）、GTPBP2过表达载体（pcDNA3.1-GTPBP2）、空载体vector（pcDNA3.1-Vector）转染HCT116细胞，分为空白组、mimics-NC+pcDNA3.1-Vector组、miR-145-5p mimics组、pcDNA3.1-GTPBP2组和pcDNA3.1-GTPBP2+miR-145-5p mimics组。采用qPCR法检测各组细胞miR-145-5p、GTPBP2 mRNA的表达水平，Western blot法检测各组细胞GTPBP2的表达水平。结果 miR-145-5p在结直肠癌组织及结直肠癌细胞中呈低表达，并且在结直肠癌组织中的表达水平与结直肠癌患者的淋巴结转移、远处转移、TNM分期相关（均P＜0.05）；GTPBP2 mRNA在结直肠癌组织及结直肠癌细胞中呈高表达；GTPBP2在结直肠癌细胞中呈高表达。miR-145-5p和GTPBP2 mRNA在结直肠癌组织中表达呈负相关（r=-0.5028，P=0.0005）；双荧光素酶报告基因实验提示miR-145-5p和GTPBP2基因之间存在靶向关系。过表达miR-145-5p可上调HCT116细胞中miR-145-5p表达，而下调GTPBP2 mRNA及GTPBP2表达。结论 miR-145-5p在结直肠癌中低表达，过表达miR-145-5p可下调HCT116细胞中GTPBP2 mRNA及GTPBP2表达水平，提示miR-145-5p可能通过靶向调控GTPBP2表达影响结直肠癌的发生和发展。 |
| 关键词: ［］ 结直肠癌 HCT116细胞 miR-145-5p GTPBP2 表达水平 |
| DOI： |
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| 基金项目:河南省医学科技攻关计划联合共建项目(LHGJ20221053) |
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| The study on the expression of miR-145-5p in colorectal cancer and its regulation mechanism on GTPBP2 gene |
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wangyongfu1,2,3,2,4
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1.The First People&2.#39;3.&4.s Hospital of Shangqiu City, Affiliated to Xinxiang Medical university
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| Abstract: |
| Abstract] Objective To explore the expression of miR-145-5p in colorectal cancer tissues and cells, as well as its mechanism of regulating colorectal cancer through the GTP binding protein 2 (GTPBP2). Methods The real-time quantitative PCR (qPCR) was used to detect the expression levels of miR-145-5p and GTPBP2 mRNA in 44 colorectal cancer tissues and their corresponding paracancerous tissues, colorectal cancer cells（HCT116）, and human normal colorectal mucosa cells(FHC）. The dual luciferase reporter gene experiments were used to verify the targeting relationship between miR-145-5p and GTPBP2 gene. miR-145-5p mimics and its negative control（mimic-NC） , GTPBP2 overexpression plasmid vector (pcDNA3.1-GTPBP2) and its blank vector (pcDNA3.1-Vector) were transfected into HCT116 cells, and divided into blank group, mimics-NC+pcDNA3.1-Vector group, miR-145-5p mimics group, pcDNA3.1-GTPBP2 group and pcDNA3.1-GTPBP2+miR-145-5p mimics group.The qPCR was used to detect the expression levels of miR-145-5p and GTPBP2 mRNA in each group, and Western blot was used to detect the expression level of GTPBP2 in each group. Results The expression level of miR-145-5p in colorectal cancer tissues and cells was significantly lower than that in paracancerous tissues and human normal colorectal mucosa cellss , and its expression level in colorectal cancer tissues was related to lymph node metastasis, distant metastasis, TNM stage in colorectal cancer patients (all P < 0.05).The expression level of GTPBP2 mRNA in colorectal cancer tissues and cells was significantly higher than that in paracancerous tissues and human normal colorectal mucosa cellss ; The expression level of GTPBP2 in colorectal cancer cells was significantly higher than that in human normal colorectal mucosa cellss . The expression level of miR-145-5p and GTPBP2 mRNA in colorectal cancer tissues showed negative correlation (r = -0.5028, P = 0.0005).The dual luciferase reporter gene experiments confirmed the targeting relationship between miR-145-5p and GTPBP2. Overexpression of miR-145-5p could upregulate the expression level of miR-145-5p in HCT116 cells, while the expression level of GTPBP2 mRNA and GTPBP2 were downregulated. Conclusion The expression level of miR-145-5p is low in colorectal cancer. Overexpression of miR-145-5p can downregulate the expression level of GTPBP2 mRNA and GTPBP2,suggesting that miR-145-5p may affect the occurrence and development of colorectal cancer by regulating GTPBP2 expression. |
| Key words: Colorectal cancer HCT116 cell miR-145-5p GTPBP2 expression level |
