The Affiliated Xinghua People's Hospital, Medical School of Yangzhou University, Yangzhou, Jiangsu, China.
Anal Cell Pathol (Amst). 2020 Dec 17;2020:4182092. doi: 10.1155/2020/4182092. eCollection 2020.
To investigate the expression of long noncoding RNA lysyl oxidase-like 1-antisense 1 (LOXL1-AS1) in hepatocellular carcinoma tissues and its effect on cell proliferation, migration, and invasion.
Quantitative real-time PCR was used to analyze the expression of LOXL1-AS1 RNA in tumor tissues, adjacent normal tissues, and cell lines. MTT assay, colony formation assay, flow cytometry analysis, transwell assays, and lentivirus-mediated RNA interference (RNAi) technology were used to evaluate cell proliferation and migration.
In the present study, we observed that the expression level of LOXL1-AS1 in hepatocellular carcinoma tissue was significantly higher than that in adjacent nontumor tissues, and its expression in three hepatic carcinoma cell lines was obviously higher than that in a normal cell line. In addition, in the Hep-G2 cell line, LOXL1-AS1 downregulation significantly inhibited cell proliferation in the light of the MTT and colony formation assays in vitro, which was consistent with animal experiment in vivo. What is more, cell migration was also inhibited in vitro in Matrigel Transwell Assay by LOXL1-AS1 knockdown, which might be partly attributed to the reduction of MMP-2 and MMP-9 protein expressions. Finally, cell cycle analysis revealed that knockdown of LOXL1-AS1 induced significantly a G0/G1 phase cell cycle arrest, which might be partly attributed to the downregulation of Cdc2, Cdc25A, and cyclin B1 protein expression.
In conclusion, we demonstrated that reduced LOXL1-AS1 expression could inhibit hepatocellular carcinoma cell proliferation, migration, and invasion. The application of RNAi targeting LOXL1-AS1 might be a potential treatment strategy in advanced cases.
探讨赖氨酰氧化酶样蛋白 1 反义 RNA1(LOXL1-AS1)在肝癌组织中的表达及其对细胞增殖、迁移和侵袭的影响。
采用实时定量 PCR 分析肿瘤组织、相邻正常组织和细胞系中 LOXL1-AS1 RNA 的表达。MTT assay、集落形成 assay、流式细胞术分析、Transwell assay 和慢病毒介导的 RNA 干扰(RNAi)技术用于评估细胞增殖和迁移。
在本研究中,我们观察到肝癌组织中 LOXL1-AS1 的表达水平明显高于相邻非肿瘤组织,且在三种肝癌细胞系中的表达明显高于正常细胞系。此外,在 Hep-G2 细胞系中,LOXL1-AS1 下调显著抑制了体外 MTT 和集落形成实验中的细胞增殖,这与体内动物实验结果一致。更重要的是,LOXL1-AS1 敲低在 Matrigel Transwell 实验中也抑制了细胞迁移,这可能部分归因于 MMP-2 和 MMP-9 蛋白表达的降低。最后,细胞周期分析显示,LOXL1-AS1 敲低诱导了明显的 G0/G1 期细胞周期停滞,这可能部分归因于 Cdc2、Cdc25A 和 cyclin B1 蛋白表达的下调。
总之,我们证明了降低 LOXL1-AS1 表达可以抑制肝癌细胞的增殖、迁移和侵袭。靶向 LOXL1-AS1 的 RNAi 应用可能是晚期病例的一种潜在治疗策略。