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非小细胞肺癌患者肿瘤组织和外周血样本中上调的miRNA-182-5p表达与下调的半胱天冬酶2表达相关。

Upregulated miRNA-182-5p expression in tumor tissue and peripheral blood samples from patients with non-small cell lung cancer is associated with downregulated Caspase 2 expression.

作者信息

Yang Lu, Dou Ye, Sui Zhuxin, Cheng Hui, Liu Xia, Wang Qinglu, Gao Peifu, Qu Yin'e, Xu Ming

机构信息

Department of Pathology and Pathophysiology, Qilu Medical University, Zibo, Shandong 255213, P.R. China.

Department of Human Anatomy, Histology and Embryology, Teaching Department of Basic Medicine, Qilu Medical University, Zibo, Shandong 255213, P.R. China.

出版信息

Exp Ther Med. 2020 Jan;19(1):603-610. doi: 10.3892/etm.2019.8074. Epub 2019 Oct 2.

Abstract

Lung cancer has the highest morbidity and mortality rates among all malignant tumors worldwide. Previous studies demonstrated that microRNA (miR)-182-5p may serve different roles in different types of cancer, including renal cell carcinoma and liver cancer. However, the functional role of miR-182-5p in non-small cell lung cancer (NSCLC) remains unknown. In the current study, the expression level of miR-182-5p in tumor tissue and peripheral blood samples obtained from patients with NSCLC was examined. The biological function of miR-182-5p on NSCLC cell proliferation was also investigated. Tissue and adjacent normal tissue samples were collected from 33 patients with NSCLC. In addition, peripheral blood samples were obtained from patients with NSCLC and 26 healthy control patients. The NSCLC cell line H1299 was used for all functional assays. Reverse transcription-quantitative polymerase chain reaction was used to determine the miR-182-5p or Caspase 2 (CASP2) mRNA expression levels in NSCLC tissue and peripheral blood samples, as well as in the NSCLC cell line. Western blotting was used to examine the protein expression level of CASP2 in tissue samples and cells, and ELISA was performed to measure the protein level of CASP2 in peripheral blood samples. MTT assay was performed to examine NSCLC cell proliferation. Flow cytometry was used to detect apoptosis. Dual-luciferase reporter assay was used to examine whether miRN182-5p directly interacts with CASP2. The current study demonstrated that miR-182-5p expression was upregulated in NSCLC tissue and peripheral blood samples from patients with NSCLC, which suggests that miR-182-5p, may serve a functional role in NSCLC. In addition, inhibition of miR-182-5p expression suppressed cell proliferation and enhanced cell apoptosis in NSCLC cells. CASP2 expression was downregulated in NSCLC tissue and peripheral blood samples from patients with NSCLC. The current study demonstrated that miR-182-5p may regulate NSCLC cell proliferation and apoptosis by regulating CASP2 expression as miR-182-5p directly binds with the 3'-untranslated region of CASP2, thereby regulating CASP2 expression.

摘要

在全球所有恶性肿瘤中,肺癌的发病率和死亡率最高。先前的研究表明,微小RNA(miR)-182-5p在不同类型的癌症中可能发挥不同作用,包括肾细胞癌和肝癌。然而,miR-182-5p在非小细胞肺癌(NSCLC)中的功能作用仍不清楚。在本研究中,检测了NSCLC患者肿瘤组织和外周血样本中miR-182-5p的表达水平。还研究了miR-182-5p对NSCLC细胞增殖的生物学功能。收集了33例NSCLC患者的组织及相邻正常组织样本。此外,还获取了NSCLC患者和26例健康对照患者的外周血样本。所有功能试验均使用NSCLC细胞系H1299。采用逆转录-定量聚合酶链反应来测定NSCLC组织、外周血样本以及NSCLC细胞系中miR-182-5p或半胱天冬酶2(CASP2)mRNA的表达水平。采用蛋白质印迹法检测组织样本和细胞中CASP2的蛋白表达水平,并采用酶联免疫吸附测定法测量外周血样本中CASP2的蛋白水平。采用MTT法检测NSCLC细胞增殖情况。采用流式细胞术检测细胞凋亡情况。采用双荧光素酶报告基因测定法检测miR-182-5p是否直接与CASP2相互作用。本研究表明,NSCLC患者的肿瘤组织和外周血样本中miR-182-5p表达上调,这表明miR-182-5p可能在NSCLC中发挥功能作用。此外,抑制miR-182-5p表达可抑制NSCLC细胞增殖并增强细胞凋亡。NSCLC患者的肿瘤组织和外周血样本中CASP2表达下调。本研究表明,miR-182-5p可能通过调节CASP2表达来调控NSCLC细胞增殖和凋亡,因为miR-182-5p直接与CASP2的3'-非翻译区结合,从而调控CASP2表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/499d/6923754/25023560a339/etm-19-01-0603-g00.jpg

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