中国癌症杂志 ›› 2021, Vol. 31 ›› Issue (5): 390-396.doi: 10.19401/j.cnki.1007-3639.2021.05.005

• 论著 • 上一篇    下一篇

miR-19b-3p靶向MTUS1调控甲状腺髓样癌细胞凋亡的分子机制

陈 国 1 ,韩鹏黎 2 ,陈柯茹 2 ,李明闯 1 ,张青松 1 ,陈 征 1 ,董汉华 1 ,钱跃军 1 ,吕 晶 1   

  1. 1. 郑州大学附属郑州中心医院甲状腺外科,河南 郑州 450007 ;
    2. 郑州大学附属郑州中心医院转化医学中心,河南 郑州 450007
  • 出版日期:2021-05-30 发布日期:2021-05-31
  • 通信作者: 吕 晶 E-mail: 903909426@qq.com
  • 基金资助:
    河南省科技攻关项目(182102310233)。

Molecular mechanism of miR-19b-3p regulating apoptosis of medullary thyroid cancer cells by targeting MTUS1

CHEN Guo 1 , HAN Pengli 2 , CHEN Keru 2 , LI Mingchuang 1 , ZHANG Qingsong 1 , CHEN Zheng 1 , DONG Hanhua 1 , QIAN Yuejun 1 , LÜ Jing #br#   

  1. 1. Department of Thyroid Surgery, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450007, Henan Province, China; 2. Translational Medicine Center, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450007, Henan Province, China
  • Published:2021-05-30 Online:2021-05-31
  • Contact: LÜ Jing E-mail: 903909426@qq.com

摘要: 背景与目的:miR-19b-3p在胃癌、前列腺癌等恶性肿瘤中均呈高表达,其可能作为肿瘤早期诊断及评估患者预后的潜在生物标志物。利用TargetScan靶基因预测软件筛选出的微管相关肿瘤抑制基因1(microtubule-associated tumor suppressor gene 1,MTUS1)可能为miR-19b-3p的靶基因,发现MTUS1的3’非翻译区(3’-untranslated region,3’-UTR)可能是miR-19b-3p的结合位点。探讨miR-19b-3p是否通过靶向MTUS1表达影响甲状腺髓样癌(medullary thyroid cancer,MTC)细胞凋亡。方法:MTC-TT细胞分为anti-miR-19b-3p组、anti-miR-NC组、pcDNA-MTUS1组、pcDNA组、anti-miR-19b-3p+si-MTUS1组、anti-miR-19b-3p+si-NC组、miR-19b-3p组和miR-NC组。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)分别检测miR-19b-3p和MTUS1的表达水平,采用双荧光素酶报告基因实验验证MTUS1是否为miR-19b-3p的靶基因;采用Western blot检测细胞中Bcl-2、Bax和cleaved caspase-3的表达水平。结果:miR-19b-3p在MTC组织及细胞系中的表达水平显著升高,而MTUS1的表达水平显著降低。转染anti-miR-19b-3p或pcDNA-MTUS1后可显著增强MTC细胞凋亡能力,促进Bax、cleaved caspase-3的表达,抑制Bcl-2的表达。双荧光素酶报告实验证实MTUS1是miR-19b-3p的靶基因。抑制MTUS1表达逆转了抑制miR-19b-3p表达对MTC-TT细胞凋亡的作用。结论:沉默miR-19b-3p表达可上调MTUS1表达从而诱导MTC细胞凋亡。

关键词: miR-19b-3p, 微管相关肿瘤抑制基因1, 甲状腺髓样癌, 凋亡

Abstract: Background and purpose: miR-19b-3p is highly expressed in malignant tumors such as gastric cancer and prostate cancer, which may be used as a potential biomarker for early diagnosis of tumor and evaluation of prognosis of patients. The microtubule-associated tumor suppressor gene 1 (MTUS1) may be miR-19b-3p target gene screened by using TargetScan target gene prediction software, and the 3’-untranslated region (3’-UTR) of MTUS1 may be the binding site of miR-19b-3p. This study aimed to investigate whether miR-19b-3p regulating the expression of MTUS1 and its effect on apoptosis of medullary thyroid cancer (MTC) cells. Methods: MTC-TT cells were divided into anti-miR-19b-3p group, anti-miR-NC group, pcDNA-MTUS1 group, pcDNA group, anti-miR-19b-3p+si-MTUS1 group, anti-miR-19b-3p+si-NC group, miR-19b-3p group and miR-NC group. Expression of miR-19b-3p and MTUS1 were detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Double luciferase reporter gene assay was performed to confirm whether MTUS1 was the target gene of miR-19b-3p. The expressions of Bcl-2, Bax and cleaved caspase-3 in cells were detected by Western blot. Results: Expression of miR-19b-3p was significantly increased in MTC tissues and cell lines, while the expression level of MTUS1 was significantly decreased. Transfection of anti-miR-19b-3p or pcDNA-MTUS1 significantly promoted the expression of Bax and cleaved caspase-3, while inhibiting Bcl-2 expression. The dual luciferase reporter assay confirmed that MTUS1 was target gene of miR-19b-3p. Inhibition of MTUS1 expression reversed the effect of inhibition of miR-19b-3p expression on apoptosis of MTC-TT cells. Conclusion: Silencing miR-19b-3p can induce apoptosis of MTC TT cells by up-regulating MTUS1 expression.

Key words: miR-19b-3p, Microtubule-associated tumor suppressor gene 1, Medullary thyroid cancer, Apoptosis