Tang Shu-Juan, Yang Jing-Bo
Department of Gastroenterology, the Second Xiangya Hospital of Central South University, No. 139, Mid Renmin Road, Furong District, Changsha, Hunan, 410011, People's Republic of China.
Mol Cell Biochem. 2020 Oct;473(1-2):143-153. doi: 10.1007/s11010-020-03815-6. Epub 2020 Jun 30.
Recurrence and adverse events after hepatocellular carcinoma (HCC) treatment occur frequently even treated with the most efficient therapy for HCC, liver transplantation. Therefore, better understanding of HCC progression is required to advance the therapeutic strategy of HCC. This study aims to explore the effect and mechanism of small nucleolar RNA host gene 14 (SNHG14) on HCC cell invasion and migration. SNHG14 and miR-656-3p expression in HCC tissues and cells were examined by qRT-PCR. After co-transfection with sh-SNHG14, miR-656-3p inhibitor, miR-656-3p mimic, si-SIRT5, pcDNA3.1-SIRT5 and corresponding negative controls, HepG2 and MHCC97H cell proliferation, invasion and migration were detected. Then the expression levels of SNHG14, miR-656-3p and SIRT5 were measured by qRT-PCR and Western blot. Luciferases reporter gene assay and RNA pull down identified the relation between SNHG14 and miR-656-3p and between miR-656-3p and SIRT5. SNHG14 was upregulated and miR-656-3p was downregulated in HCC cells. Inhibition of SNHG14 could inhibit HepG2 and MHCC97H cell proliferation, invasion and migration. Upregulation of miR-656-3p or knockdown of SIRT5 significantly suppressed the biological process of HepG2 and MHCC97H cells. SNHG14 directly acted on miR-656-3p and SIRT5 was a target gene of miR-656-3p. miR-656-3p inhibitor or pcDNA3.1-SIRT5 could reverse the inhibition of sh-SNHG14 on cell proliferation, invasion and migration of HCC cells. SNHG14 promotes HCC cell invasion and migration through regulating miR-656-3p/SIRT5 axis.
即使采用肝癌最有效的治疗方法——肝移植进行治疗,肝细胞癌(HCC)治疗后的复发和不良事件仍频繁发生。因此,需要更好地了解HCC的进展情况,以推进HCC的治疗策略。本研究旨在探讨小核仁RNA宿主基因14(SNHG14)对HCC细胞侵袭和迁移的影响及机制。通过qRT-PCR检测HCC组织和细胞中SNHG14和miR-656-3p的表达。在与sh-SNHG14、miR-656-3p抑制剂、miR-656-3p模拟物、si-SIRT5、pcDNA3.1-SIRT5及相应阴性对照共转染后,检测HepG2和MHCC97H细胞的增殖、侵袭和迁移情况。然后通过qRT-PCR和蛋白质免疫印迹法检测SNHG14、miR-656-3p和SIRT5的表达水平。荧光素酶报告基因检测和RNA下拉实验确定了SNHG14与miR-656-3p之间以及miR-656-3p与SIRT5之间的关系。HCC细胞中SNHG14上调而miR-656-3p下调。抑制SNHG14可抑制HepG2和MHCC97H细胞的增殖、侵袭和迁移。上调miR-656-3p或敲低SIRT5可显著抑制HepG2和MHCC97H细胞的生物学过程。SNHG14直接作用于miR-656-3p,且SIRT5是miR-656-3p的靶基因。miR-656-3p抑制剂或pcDNA3.1-SIRT5可逆转sh-SNHG14对HCC细胞增殖、侵袭和迁移的抑制作用。SNHG14通过调节miR-656-3p/SIRT5轴促进HCC细胞的侵袭和迁移。