Lin Rui-Xin, Zhan Guo-Feng, Wu Jia-Cheng, Fang He, Yang Shu-Li
Department of Hepato-Biliary-Pancreatic Surgery, The Second Hospital of Jilin University, Changchun, 130041, People's Republic of China.
Department of Obstetrics and Gynecology, The Second Hospital of Jilin University, No. 218, Ziqiang Street, Changchun, 130041, People's Republic of China.
Dig Dis Sci. 2022 Mar;67(3):936-946. doi: 10.1007/s10620-021-06920-8. Epub 2021 Mar 29.
To explore how lncRNA SNHG14 modulates the biological features of hepatocellular carcinoma (HCC) cells by regulating SOX9 via mediating miR-206.
HCC tissues were collected to perform the quantitative reverse transcriptase polymerase chain reaction to determine the expressions of SNHG14, miR-206, and SOX9. HCC cell line SMCC7721 was selected for co-transfection by si-SNHG14/miR-206 inhibitor/si-SOX9, followed by the measurement of cell proliferation using Cell Count Kit-8 (CCK-8) assay and clone formation assay. The migration and invasion were evaluated by wound healing test and Transwell assay. The apoptotic rate was determined by flow cytometry. Levels of the apoptosis-related proteins were measured through Western blotting.
SNHG14 and SOX9 were up-regulated in HCC tumor tissues compared with adjacent normal tissues, with decreased miR-206 expression. Moreover, SNHG14 expression was significantly associated with the TNM stage, lymphatic metastasis, and histological differentiation of HCC patients. Besides, inverse correlations between SNHG14 and miR-206, as well as between miR-206 and SOX9, were noted. The dual luciferase reporter gene assay, RIP, and RNA pull-down experiments also revealed the targeting relationship between SNHG14 and miR-206 or between miR-206 and SOX9. Silencing SNHG14 and SOX9 inhibited the proliferation, invasion, and migration of HCC cells, with increased apoptosis, which was all abolished by silencing miR-206.
Inhibition of SNHG14 suppresses SOX9 by up-regulating miR-206, to further inhibit the proliferation, migration, and invasion of HCC cells with the promoted apoptosis, which is a novel target for the treatment of HCC.
探讨长链非编码RNA SNHG14如何通过介导miR-206调控SOX9来调节肝癌（HCC）细胞的生物学特性。
收集HCC组织，进行定量逆转录聚合酶链反应以测定SNHG14、miR-206和SOX9的表达。选择HCC细胞系SMCC7721进行si-SNHG14/miR-206抑制剂/si-SOX9共转染，随后使用细胞计数试剂盒-8（CCK-8）法和克隆形成试验测量细胞增殖。通过伤口愈合试验和Transwell试验评估迁移和侵袭能力。通过流式细胞术测定凋亡率。通过蛋白质印迹法测量凋亡相关蛋白的水平。
与相邻正常组织相比，HCC肿瘤组织中SNHG14和SOX9上调，miR-206表达降低。此外，SNHG14表达与HCC患者的TNM分期、淋巴结转移和组织学分化显著相关。此外，还发现SNHG14与miR-206之间以及miR-206与SOX9之间呈负相关。双荧光素酶报告基因试验、RNA免疫沉淀和RNA下拉实验也揭示了SNHG14与miR-206之间或miR-206与SOX9之间的靶向关系。沉默SNHG14和SOX9可抑制HCC细胞的增殖、侵袭和迁移，同时增加凋亡，而沉默miR-206可消除所有这些作用。
抑制SNHG14可通过上调miR-206抑制SOX9，从而进一步抑制HCC细胞的增殖、迁移和侵袭，并促进凋亡，这是治疗HCC的一个新靶点。
