Institute of Liver Disease, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China.
Department of Liver Disease, Hospital for Infectious Diseases of Pudong District, Shanghai 201299, P.R. China.
Mol Med Rep. 2022 Apr;25(4). doi: 10.3892/mmr.2022.12628. Epub 2022 Feb 4.
The aim of the present study was to investigate the molecular mechanisms of atractylon in the inhibition of invasion and migration of hepatic cancer cells. High‑throughput sequencing was used to compare the expression of long non‑coding (lnc)RNAs between hepatic carcinoma and healthy controls. A competing endogenous RNA network was constructed. The top significantly differentially expressed lncRNAs were screened and verified by reverse transcription‑quantitative PCR and . Small interfering (si)RNA against thymopoietin‑antisense 1 (TMPO‑AS1) or coiled‑coil domain‑containing 183‑antisense 1 (CCDC183‑AS1) overexpression (oe) vectors were transfected into cells following atractylon treatment. Wound healing and Matrigel assays were used to determine the effects of migration and invasion, respectively. Western blot analysis was used to detect the expression levels of invasion‑ and migration‑related proteins, including N‑cadherin, E‑cadherin and MMP‑2. Flow cytometry analysis was used to detect apoptosis. Based on transcriptome sequencing and analysis, the top seven upregulated [(FAM201A, RP11‑640M9.2, AL589743.1, TMEM51‑AS1, clathrin heavy chain‑like 1 (CLTCL1), TMPO‑AS1 and LINC00652] and top six downregulated lncRNAs (RP11‑465B22.5, CCDC183‑AS1, TCONS_00072529, RP11‑401F2.3, RP11‑290F20.1 and TCONS_00070568) were identified. Only TMPO‑AS1 and CCDC183‑AS1 were differently regulated by atractylon . The proliferative ability of HepG2 liver cancer cells decreased, whereas the apoptotic rate improved after atractylon treatment. Notably, the invasive and migratory ability of HepG2 cells significantly declined. In addition, siTMPO‑AS1 and oeCCDC183‑AS1 reduced the effect of atractylon in vitro. Atractylon was demonstrated to regulate the expression of TMPO‑AS1 and CCDC183‑AS1 and inhibited the invasion and migration of liver cancer cells. Thus, TMPO‑AS1 and CCDC183‑AS1 may be potential targets for diagnosis and treatment of hepatic carcinoma.
本研究旨在探讨吴茱萸次碱抑制肝癌细胞侵袭和迁移的分子机制。采用高通量测序比较肝癌与健康对照组织中长链非编码(lnc)RNA 的表达。构建竞争内源性 RNA 网络。筛选并验证差异表达最显著的 lncRNA 采用逆转录-定量 PCR 和 Western blot 分析。用小干扰(si)RNA 敲低胸腺素-β-10 反义 RNA1(TMPO-AS1)或卷曲螺旋结构域 183 反义 RNA1(CCDC183-AS1)过表达(oe)载体,转染细胞,进行吴茱萸次碱处理。划痕愈合和基质胶实验分别用于检测迁移和侵袭。Western blot 分析用于检测侵袭和迁移相关蛋白(包括 N-钙黏蛋白、E-钙黏蛋白和 MMP-2)的表达水平。流式细胞术分析用于检测细胞凋亡。基于转录组测序和分析,确定了上调最显著的 7 个 lncRNA(FAM201A、RP11-640M9.2、AL589743.1、TMEM51-AS1、网格蛋白重链样 1(CLTCL1)、TMPO-AS1 和 LINC00652)和下调最显著的 6 个 lncRNA(RP11-465B22.5、CCDC183-AS1、TCONS_00072529、RP11-401F2.3、RP11-290F20.1 和 TCONS_00070568)。只有 TMPO-AS1 和 CCDC183-AS1 被吴茱萸次碱差异调节。吴茱萸次碱处理后 HepG2 肝癌细胞的增殖能力下降,凋亡率提高。此外,siTMPO-AS1 和 oeCCDC183-AS1 降低了吴茱萸次碱在体外的作用。吴茱萸次碱被证明可以调节 TMPO-AS1 和 CCDC183-AS1 的表达,并抑制肝癌细胞的侵袭和迁移。因此,TMPO-AS1 和 CCDC183-AS1 可能是肝癌诊断和治疗的潜在靶点。