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LncRNA H19/miR-193a-3p 轴通过靶向 PSEN1 修饰肝癌细胞的放射抵抗和化疗耐受性。

The LncRNA H19/miR-193a-3p axis modifies the radio-resistance and chemotherapeutic tolerance of hepatocellular carcinoma cells by targeting PSEN1.

机构信息

Department of Radiotherapy, Eastern Hepatobiliary Surgery Hospital, ShangHai, China.

The First Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Shanghai, China.

出版信息

J Cell Biochem. 2018 Nov;119(10):8325-8335. doi: 10.1002/jcb.26883. Epub 2018 Jul 3.

Abstract

This study was designated to verify if the lncRNA H19/miR-193a-3p axis would play a regulatory role in the radio-/chemo-resistances of HCC cells through targeting PSEN1. Within the study, five human HCC cell lines were prepared, including Bel-7402, HepG2, Hep3b, QGY-7703, and SMMC-7721. Moreover, docetaxel (DT), paclitaxel (Pt), vinorelbine (Vb), and 5-fluorouracil (5-Fu) were managed as the chemo-therapeutics, and single-dose X-rays were performed as radio-therapies. Besides, lncRNA H19 and miR-193a-3p were detected by qRT-PCR and Western blot were implemented to quantify the expressional levels of PSEN1, Ku80, γ-H2AX, and RAD51. Luciferase reporter gene assay was advanced to verify the targeted relationship between lncRNA H19 and miR-193a-3p. As a consequence, QGY-7703 and Bel-7402 were, respectively, the most radiation-sensitive and radiation-proof cell lines, and Bel-7402 was associated with the highest resistances to DT, Pt, Vb, and 5-FU. The restrained lncRNA H19 and over-expressed miR-193a-3p expressions tended to significantly elevate the survival rate and proliferation of Bel-7402 cells, when they were exposed to radiation and subject to chemo-therapies. The lncRNA H19 was also found to directly target miR-193a-3p in inducing the HCC development. PSEN1 appeared to be subject to the modification of lncRNA H19 and miR-193a-3p in its acting on the survival rates and proliferative abilities of HCC cells. The lncRNA H19/miR-193a-3p/PSEN1 axis could be regarded as the treatment targets for HCC, so as to further improve the treatment efficacy of chemo- and radio-therapies for HCC.

摘要

本研究旨在验证长链非编码 RNA H19/miR-193a-3p 轴是否通过靶向 PSEN1 在 HCC 细胞的放射/化疗耐药中发挥调节作用。在这项研究中,制备了包括 Bel-7402、HepG2、Hep3b、QGY-7703 和 SMMC-7721 在内的五个人肝癌细胞系。此外,使用多西紫杉醇(DT)、紫杉醇(Pt)、长春瑞滨(Vb)和 5-氟尿嘧啶(5-Fu)作为化疗药物,单次 X 射线作为放射治疗。此外,通过 qRT-PCR 检测 lncRNA H19 和 miR-193a-3p,Western blot 用于定量 PSEN1、Ku80、γ-H2AX 和 RAD51 的表达水平。通过荧光素酶报告基因检测验证 lncRNA H19 与 miR-193a-3p 的靶向关系。结果表明,QGY-7703 和 Bel-7402 分别是最敏感和最耐药的细胞系,Bel-7402 对 DT、Pt、Vb 和 5-Fu 的耐药性最高。抑制 lncRNA H19 和过表达 miR-193a-3p 表达倾向于显著提高 Bel-7402 细胞在辐射和化疗暴露下的存活率和增殖率。还发现 lncRNA H19 通过直接靶向 miR-193a-3p 诱导 HCC 的发生。PSEN1 似乎受到 lncRNA H19 和 miR-193a-3p 修饰的影响,从而影响 HCC 细胞的存活率和增殖能力。lncRNA H19/miR-193a-3p/PSEN1 轴可作为 HCC 的治疗靶点,以进一步提高 HCC 化疗和放疗的治疗效果。

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