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miR-101对肝细胞癌预后的临床影响及抗miR-101的致癌机制

Clinical effects of miR-101 on prognosis of hepatocellular carcinoma and carcinogenic mechanism of anti-miR-101.

作者信息

Lv Xuecheng, Li Jinghua, Yang Bingnan

机构信息

Department of Pharmacy, The First People's Hospital of Shangqiu, Shangqiu, Henan 476100, P.R. China.

Department of Cardiovascular Surgery, Henan Provincial People's Hospital, Zhengzhou, Henan 450000, P.R. China.

出版信息

Oncol Rep. 2016 Oct;36(4):2184-92. doi: 10.3892/or.2016.4980. Epub 2016 Jul 28.

Abstract

The aim of this study was to verify whether anti-miR-101 participates in the treatment of hepatocellular carcinoma (HCC) as a small-molecule antitumor agent, and to explore the effect on phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Patients who received consecutive hepatectomies were followed-up, and miR-101 expressions in their tumor and paracancerous tissues were detected. Correlation between miR-101 expression and clinical pathological factors and prognosis was studied. High‑throughput sequencing was used to detect the genetic and microRNA (miRNA) levels of tumor tissues. Expression of anti-miR-101 in different HCC cell lines was determined, and those of desired genes and proteins were detected by qRT-PCR and western blotting to obtain the target gene. miR-101 was significantly upregulated in HCC patients compared with that in paracancerous tissues. High miR-101 expression, vascular invasion, tumor size ≥7 cm and late pathological stage were the risk factors of recurrence-free survival rate. High miR-101 expression was the independent prognostic factor of total and recurrence-free survival rates. CXCL12, IL6R, FOXO3 and PTEN were screened as desired genes, and only PTEN was expressed significantly differently in three cell lines. miR-101 could bind 3'-UTR of WT-PTEN with reduced fluorescent intensity, suggesting that PTEN was the target gene. SMMC-7721, HepG2 and Huh7 were eligible cell lines for miR-101 studies. miR-101 was an applicable molecular marker of HCC. Anti-miR-101 regulated the transcription of PTEN and may promote cell proliferation, differentiation and apoptosis by regulating downstream genes with PTEN. The regulatory effects of anti-miR-101 on PTEN provide valuable evidence for finding novel miRNA drugs.

摘要

本研究旨在验证抗 miR-101 作为一种小分子抗肿瘤药物是否参与肝细胞癌(HCC)的治疗,并探讨其对 10 号染色体缺失的磷酸酶和张力蛋白同源物(PTEN)的影响。对接受连续肝切除术的患者进行随访,检测其肿瘤组织和癌旁组织中 miR-101 的表达。研究 miR-101 表达与临床病理因素及预后的相关性。采用高通量测序检测肿瘤组织的基因和微小 RNA(miRNA)水平。测定抗 miR-101 在不同 HCC 细胞系中的表达,并通过 qRT-PCR 和蛋白质印迹法检测相关基因和蛋白的表达以获得靶基因。与癌旁组织相比,HCC 患者中 miR-101 显著上调。miR-101 高表达、血管侵犯、肿瘤大小≥7 cm 和病理分期较晚是无复发生存率的危险因素。miR-101 高表达是总生存率和无复发生存率的独立预后因素。筛选出 CXCL12、IL6R、FOXO3 和 PTEN 作为相关基因,其中只有 PTEN 在三种细胞系中的表达有显著差异。miR-101 可与 WT-PTEN 的 3'-UTR 结合,荧光强度降低,提示 PTEN 是靶基因。SMMC-7721、HepG2 和 Huh7 是适用于 miR-101 研究的细胞系。miR-101 是 HCC 适用的分子标志物。抗 miR-101 调节 PTEN 的转录,并可能通过调节 PTEN 的下游基因促进细胞增殖、分化和凋亡。抗 miR-101 对 PTEN 的调节作用为寻找新型 miRNA 药物提供了有价值的证据。

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