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。
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.
作者
陈国
韩鹏黎
陈柯茹
李明闯
张青松
陈征
董汉华
钱跃军
吕晶
CHEN Guo;HAN Pengli;CHEN Keru;LI Mingchuang;ZHANG Qingsong;CHEN Zheng;DONG Hanhua;QIAN Yuejun;LÜJing(Department of Thyroid Surgery,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,Henan Province,China;Translational Medicine Center,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,Henan Province,China)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2021年第5期390-396,共7页
China Oncology
基金
河南省科技攻关项目(182102310233)。
