Wang Zhenchang, Huang DanDan, Huang Jingjing, Nie Kunmei, Li Xiaofan, Yang Xiaojin
Department of Spleen and Stomach Liver Disease, International Zhuang Hospital District of Guangxi University of Chinese Medicine, Nanning, Guangxi, People's Republic of China.
Basic Medical Science College, North Sichuan Medical College, Nanchong, People's Republic of China.
Onco Targets Ther. 2020 Jul 3;13:6539-6551. doi: 10.2147/OTT.S250355. eCollection 2020.
Previous evidence have shown that long non-coding RNA (lncRNA) TMPO antisense RNA 1 (TMPO-AS1) is involved in the aggressiveness of several cancers. Nevertheless, the precise functions of TMOP-AS1 in hepatocellular carcinoma (HCC) are still unresolved.
The expressions of TMPO-AS1 and miR-320a were detected in HCC tissues and cells by qRT-RCR. The cell growth, migration and invasion were detected by colony formation, wound healing assay and Transwell assay, respectively. The targeting relation between miR-320a and TMPO-AS1 was predicted by bioinformatics analysis and identified by luciferase reporter gene as well as FISH assay. The expression of SERPINE1 MRNA Binding Protein 1 (SERBP1) was detected by Western blot. The growth of HCC cell was analyzed using transplanted tumor model.
Currently, we revealed that TMPO-AS1 was overexpressed in clinical HCC samples and a panel of HCC cell lines. Clinically, a higher level of TMPO-AS1 was connected to the advanced stage of HCC and worse prognosis of patients. Depletion of TMPO-AS1 repressed HCC cell viability, migration ability and invasiveness. Nevertheless, upregulation of TMPO-AS1 caused opposite results. Further studies revealed that lncRNA TMPO-AS1 was largely located in the cytoplasm of HCC cell and sponge miR-320a, resulting in increasing the level of SERBP1 in HCC cell. Finally, TMPO-AS1 silencing suppressed tumor growth of HCC cell in vivo.
Collectively, our results suggested that TMPO-AS1 was a promoting factor for the aggressive behaviors of HCC cell.
先前的证据表明,长链非编码RNA(lncRNA)TMPO反义RNA 1(TMPO-AS1)参与了多种癌症的侵袭性。然而,TMOP-AS1在肝细胞癌(HCC)中的具体功能仍未明确。
采用qRT-RCR检测HCC组织和细胞中TMPO-AS1和miR-320a的表达。分别通过集落形成、伤口愈合试验和Transwell试验检测细胞的生长、迁移和侵袭能力。通过生物信息学分析预测miR-320a与TMPO-AS1之间的靶向关系,并通过荧光素酶报告基因以及FISH试验进行鉴定。采用蛋白质免疫印迹法检测丝氨酸蛋白酶抑制剂1 mRNA结合蛋白1(SERBP1)的表达。利用移植瘤模型分析HCC细胞的生长情况。
目前,我们发现TMPO-AS1在临床HCC样本和一组HCC细胞系中高表达。临床上,TMPO-AS1水平较高与HCC晚期及患者预后较差相关。TMPO-AS1的缺失抑制了HCC细胞的活力、迁移能力和侵袭性。然而,TMPO-AS1的上调则产生相反的结果。进一步研究表明,lncRNA TMPO-AS1主要位于HCC细胞的细胞质中,并可吸附miR-320a,导致HCC细胞中SERBP1水平升高。最后,TMPO-AS1沉默抑制了体内HCC细胞的肿瘤生长。
总体而言,我们的结果表明TMPO-AS1是HCC细胞侵袭性行为的促进因子。