Department of Radiotherapy, Eastern Hepatobiliary Surgery Hospital, ShangHai, China.
The First Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Shanghai, China.
J Cell Biochem. 2018 Nov;119(10):8325-8335. doi: 10.1002/jcb.26883. Epub 2018 Jul 3.
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.
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。