miR-182-5p在膀胱癌中的表达及其机制

Expression of miR-182-5p and its mechanism in bladder carcinoma

  • 摘要:
      目的  探究miR-182-5p及FOXO3在膀胱癌中的表达意义及其参与膀胱癌可能的机制。
      方法  收集2017年6月至2019年6月64例于郑州人民医院行手术切除膀胱癌患者的组织标本和临床资料,采用RT-qPCR检测膀胱癌组织及膀胱癌细胞中的miR-182-5p和FOXO3表达,按照其表达水平分为miR-182-5p高表达组和miR-182-5p低表达组、FOXO3高表达组和FOXO3低表达组,采用Western blot实验检测FOXO3蛋白的表达。采用Kaplan-Meier生存曲线分析miR-182-5p和FOXO3与膀胱癌患者预后的相关性。采用双荧光素酶报告实验分析miR-182-5p与FOXO3的靶向关系,并采用实时荧光定量PCR(RT-qPCR)和Western blot检测转染miR-182-5p抑制剂对FOXO3表达的影响;分别采用CCK-8、Transwell实验和流式细胞术检测转染miR-182-5p抑制剂对T24细胞增殖、侵袭和凋亡的影响。
      结果  miR-182-5p在膀胱癌组织中的表达量为2.27±0.66,显著高于癌旁组织的1.04±0.36,两者比较差异具有统计学意义(P<0.001),与肿瘤大小、TNM分期、淋巴结转移有关(P<0.05);FOXO3在膀胱癌组织中低表达,与肿瘤大小、分化程度、TNM分期及淋巴结转移有关(P<0.05);miR-182-5p高表达组、FOXO3低表达组的患者的生存率显著低于miR-182-5p低表达组、FOXO3高表达组(P=0.038,P=0.004)。T24细胞中miR-182-5p高表达(P<0.05)。抑制miR-182-5p上调FOXO3表达,抑制miR-182-5p表达能抑制T24细胞的增殖和侵袭,促进细胞凋亡(P<0.05)。
      结论  miR-182-5p在膀胱癌组织中高表达,而FOXO3低表达。miR-182-5p促进膀胱癌细胞的增殖和侵袭,抑制其凋亡,可能与负调控FOXO3有关。

     

    Abstract:
      Objective  To investigate the expression of miR-182-5p and FOXO3 in bladder carcinoma and decipher the mechanism of it.
      Methods  The expression of miR-182-5p and FOXO3 in bladder carcinoma and adjacent tissues samples from 64 bladder cancer patients at People’s Hospital of Zhengzhou, from June 2017 to June 2019 and in cell lines were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Based on the expression of miR-182-5p or FOXO3, the patients were assigned into miR-182-5p high and miR-182-5p low groups or FOXO3 high and FOXO3 low groups, respectively. The expression of FOXO3 protein was detected by Western blot. Kaplan-Meier survival curve was used to analyze the correlation between miR-182-5p or FOXO3 and the prognosis of bladder carcinoma patients. Dual luciferase reporter assays were used to validate the targeting of FOXO3 by miR-182-5p. Further, the effect of the miR-182-5p inhibitor on the expression of FOXO3 in bladder carcinoma cells was assessed by RT-qPCR and Western blot. The role of miR-182-5p on bladder carcinoma was determined by transfecting the T24 cells with miR-182-5p inhibitor and assessing the effect on cell proliferation, invasion, and apoptosis using the CCK-8 assay, the Transwell invasion assay, and flow cytometry, respectively.
      Results  The expression of miR-182-5p in bladder cancer tissues was higher (2.27±0.66) than that in adjacent tissues (1.04±0.36), and this difference was statistically significant (P<0.001). The expression of miR-182 was linked to tumor size, TNM stage, and lymph node metastasis. On the contrary, the expression of FOXO3 in bladder cancer tissues was downregulated and was closely associated with tumor size, degree of differentiation, TNM stage, and lymph node metastasis (P<0.05). The survival rates of bladder carcinoma patients in miR-182-5p high or FOXO3 low groups were significantly lower than those in the miR-182-5p low or FOXO3 high groups (P=0.038, P=0.004), respectively. The expression of miR-182-5p in T24 cells was significantly increased. Furthermore, the miR-182-5p inhibitor upregulated the expression of FOXO3 in T24 bladder carcinoma cells. However, the downregulation of miR-182-5p inhibited proliferation and invasion and promoted apoptosis in T24 cells (P<0.05).
      Conclusion  miR-182-5p expression is upregulated and FOXO3 expression is downregulated in bladder carcinoma tissues. miR-182-5p promotes cell proliferation and invasion and inhibits apoptosis in bladder carcinoma cells, which may be attributed to the negative regulation of FOXO3, an established tumor suppressor in bladder cancer.

     

/

返回文章
返回