Molecular mechanism of atractylon in the invasion and migration of hepatic cancer cells based on high­throughput sequencing.

ABSTRACT

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. High­throughput sequencing was used to compare the expression of long non­coding (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 transcription­quantitative PCR in vitro and in vivo. Small interfering (si)RNA against thymopoietin­antisense 1 (TMPO­AS1) or coiled­coil domain­containing 183­antisense 1 (CCDC183­AS1) 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 migration­related proteins, including N­cadherin, E­cadherin and MMP­2. Flow cytometry analysis was used to detect apoptosis. Based on transcriptome sequencing and analysis, the top seven upregulated [(FAM201A, RP11­640M9.2, AL589743.1, TMEM51­AS1, clathrin heavy chain­like 1 (CLTCL1), TMPO­AS1 and LINC00652] and top six downregulated lncRNAs (RP11­465B22.5, CCDC183­AS1, TCONS_00072529, RP11­401F2.3, RP11­290F20.1 and TCONS_00070568) were identified. Only TMPO­AS1 and CCDC183­AS1 were differently regulated by atractylon in vivo. 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, siTMPO­AS1 and oeCCDC183­AS1 reduced the effect of atractylon in vitro. Atractylon was demonstrated to regulate the expression of TMPO­AS1 and CCDC183­AS1 and inhibited the invasion and migration of liver cancer cells. Thus, TMPO­AS1 and CCDC183­AS1 may be potential targets for diagnosis and treatment of hepatic carcinoma.