Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, PR China.
Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, PR China.
Kaohsiung J Med Sci. 2018 Oct;34(10):547-555. doi: 10.1016/j.kjms.2018.05.003. Epub 2018 Jun 12.
Hepatocellular carcinoma (HCC) is a frequently occurred malignancy worldwide with a high mortality. The treatment for HCC is still controversial. Emerging evidences have demonstrated that microRNAs (miRs) play a role in HCC. This study aims to investigate the effects of lentiviral-mediated miRNA-26b (miR-26b) on the proliferation and metastasis of HCC cells. The normal hepatic cell line HL-7702 and HCC cell lines HepG2 (without metastatic potential), SMMC-7721 (with low metastatic potential) and MHCC97H (with high metastatic potential) were purchased for our experiment. The lentiviral-mediated miR-26b overexpression (miR-26b-LV) and low expression (sh-miR-26b) were constructed to transfect the cells. The miR-26b expression and expressions of Karyopherin α-2 (KPNA2), matrix metalloproteinase 1 (MMP-1), MMP-7 and MMP-14 were determined by RT-qPCR and western blot analysis. The proliferation and metastasis of transfected HCC cells were detected by MTT and Transwell assay respectively. The miR-26b expressions were decreased significantly in MHCC97H cells. With lentiviral-mediated miR-26b overexpression, the proliferation and migration of HepG2, MHCC97H and SMMC-7721 cells were decreased significantly. The RT-qPCR and western blot analysis results revealed that the mRNA and protein expressions of KPNA2, MMP-1, MMP-7 and MMP-14 were decreased by lentiviral-mediated miR-26b overexpression. All the above indexes in the HepG2, MHCC97H and SMMC-7721 cells treated by sh-miR-26b exhibited opposite trends. These results show that overexpressed miR-26b could inhibit the proliferation and metastasis of HCC cells significantly, which provides a novel target and theoretical foundation for the treatment of HCC.
肝细胞癌（HCC）是一种全球范围内常见的恶性肿瘤，死亡率较高。HCC 的治疗仍存在争议。新出现的证据表明 microRNAs（miRs）在 HCC 中发挥作用。本研究旨在探讨慢病毒介导的 miRNA-26b（miR-26b）对 HCC 细胞增殖和转移的影响。本实验购买了正常肝细胞系 HL-7702 和 HCC 细胞系 HepG2（无转移潜能）、SMMC-7721（低转移潜能）和 MHCC97H（高转移潜能）。构建了慢病毒介导的 miR-26b 过表达（miR-26b-LV）和低表达（sh-miR-26b）转染细胞。通过 RT-qPCR 和 Western blot 分析测定 miR-26b 表达及核孔蛋白α-2（KPNA2）、基质金属蛋白酶 1（MMP-1）、MMP-7 和 MMP-14 的表达。通过 MTT 和 Transwell 测定转染 HCC 细胞的增殖和转移。MHCC97H 细胞中 miR-26b 表达明显降低。通过慢病毒介导的 miR-26b 过表达，HepG2、MHCC97H 和 SMMC-7721 细胞的增殖和迁移明显减少。RT-qPCR 和 Western blot 分析结果显示，慢病毒介导的 miR-26b 过表达降低了 KPNA2、MMP-1、MMP-7 和 MMP-14 的 mRNA 和蛋白表达。sh-miR-26b 处理的 HepG2、MHCC97H 和 SMMC-7721 细胞的所有上述指标均表现出相反的趋势。这些结果表明，过表达 miR-26b 可显著抑制 HCC 细胞的增殖和转移，为 HCC 的治疗提供了新的靶点和理论基础。
