引用本文:王永富,郑紫恒,许春进.miR-145-5p在结直肠癌中的表达及对GTPBP2基因靶向调控的研究[J].中国临床新医学,2024,17(5):511-516.
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miR-145-5p在结直肠癌中的表达及对GTPBP2基因靶向调控的研究
王永富,郑紫恒,许春进
新乡医学院附属商丘市第一人民医院消化内科,河南 476100
摘要:
[摘要] 目的 探讨miR-145-5p在结直肠癌中的表达及对GTP结合蛋白2(GTPBP2)的调控作用。方法 选择2022年1月至12月在新乡医学院附属商丘市第一人民医院行结直肠癌根治性切除术的44例结直肠癌患者的癌组织及癌旁组织。选择人正常结直肠黏膜细胞FHC、结直肠癌细胞HCT116进行基础实验。采用荧光定量聚合酶链反应(qPCR)检测癌组织、癌旁组织及细胞的miR-145-5p和GTPBP2 mRNA的表达水平。采用Western blot法检测细胞GTPBP2蛋白表达水平。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。分析结直肠癌组织miR-145-5p表达水平与患者临床病理特征的关联性。结果 与癌旁组织相比,癌组织中miR-145-5p表达水平显著降低(t=8.189,P<0.001),GTPBP2 mRNA表达水平显著升高(t=11.230,P<0.001)。Pearson相关性分析结果显示,结直肠癌组织miR-145-5p表达水平与GTPBP2 mRNA表达水平呈负相关(r=-0.503,P<0.001)。miR-145-5p表达水平与结直肠癌远处转移、淋巴结转移以及TNM分期存在显著关联(P<0.05),但与患者性别、年龄、病灶部位、临床T分期、肿瘤直径的关联性不显著(P>0.05)。与FHC细胞相比,HCT116细胞miR-145-5p表达水平显著降低(P<0.001),GTPBP2 mRNA和蛋白表达水平显著升高(P<0.001)。过表达miR-145-5p可下调HCT116细胞GTPBP2 mRNA和蛋白的表达。双荧光素酶报告基因实验结果提示miR-145-5p和GTPBP2基因之间存在靶向关系。结论 miR-145-5p在结直肠癌中呈低表达,并与远处转移、淋巴结转移和TNM分期存在显著关联,其可能通过靶向调控GTPBP2表达而影响结直肠癌的发生、发展。
关键词:  结直肠癌  miR-145-5p  GTP结合蛋白2  靶向调控
DOI:10.3969/j.issn.1674-3806.2024.05.07
分类号:
基金项目:河南省医学科技攻关计划联合共建项目(编号:LHGJ20221053)
A study on expression of miR-145-5p in colorectal cancer and its targeted regulation on GTPBP2 gene
WANG Yongfu, ZHENG Ziheng, XU Chunjin
Department of Gastroenterology, Shangqiu First People′s Hospital, Xinxiang Medical College, Henan 476100, China
Abstract:
[Abstract] Objective To explore the expression of miR-145-5p in colorectal cancer and its regulatory effect on guanosine triphosphate(GTP)-binding protein 2(GTPBP2). Methods The cancer tissues and cancer-adjacent tissues of 44 patients with colorectal cancer who underwent radical resection of colorectal cancer in Shangqiu First People′s Hospital, Xinxiang Medical College from January 2022 to December 2022 were selected. Normal human colorectal mucosa cell FHC and colorectal cancer cell HCT116 were selected for basic experiments. Fluorescent quantitative polymerase chain reaction(qPCR) was used to detect the expression levels of miR-145-5p and GTPBP2 mRNA in cancer tissues, cancer-adjacent tissues and the cells. Western blot method was used to detect the expression level of GTPBP2 protein in the cells. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-145-5p and GTPBP2 gene. The correlation between the expression level of miR-145-5p in colorectal cancer tissues and the clinicopathological features of the patients was analyzed. Results Compared with that in the cancer-adjacent tissues, the expression level of miR-145-5p in cancer tissues was significantly decreased(t=8.189, P<0.001), and the mRNA expression level of GTPBP2 was significantly increased(t=11.230, P<0.001). The results of Pearson correlation analysis showed that the expression level of miR-145-5p in colorectal cancer tissues was negatively correlated with the mRNA expression level of GTPBP2(r=-0.503, P<0.001). The expression level of miR-145-5p was significantly associated with distant metastasis, lymph node metastasis and TNM stage of colorectal cancer(P<0.05), but was not significantly associated with the patients′ gender, age, lesion location, clinical T stage and tumor diameter(P>0.05). Compared with that in the FHC cells, the expression level of miR-145-5p in the HCT116 cells was significantly decreased(P<0.001), and the mRNA and protein expression levels of GTPBP2 were significantly increased(P<0.001). Overexpression of miR-145-5p could down-regulate the expressions of GTPBP2 mRNA and protein in the HCT116 cells. The results of dual luciferase reporter gene assay suggested a targeting relationship between miR-145-5p and GTPBP2 gene. Conclusion The expression of miR-145-5p is low in colorectal cancer and miR-145-5p is significantly associated with distant metastasis, lymph node metastasis, and TNM stage and it may affect the occurrence and development of colorectal cancer through targeted regulation of GTPBP2 expression.
Key words:  Colorectal cancer  miR-145-5p  Guanosine triphosphate(GTP)-binding protein 2(GTPBP2)  Targeted regulation