Long-chain non-coding RNA ZNF674-AS1 regulates proliferation and apoptosis of hepatoma cells via Wnt signaling pathway

Objective To detect the expression of long-chain non-coding RNA ZNF674-AS1 in hepatocarcinoma and explore its effect on the proliferation and apoptosis of hepatocellular carcinoma cells. Methods Clinical liver cancer specimens (10 cases) and paracancerous specimens (10 cases) were collected and the expression level of ZNF674-AS1 in tissues was quantified by reverse transcriptase-polymerase chain reaction (RT-PCR). A ZNF674-AS1 stable knockout cell line was constructed by small interfering RNA (siRNA) in hepatocellular carcinoma cell lines HepG2 and QGY7701, and cell counting kit-8 (CCK-8) kit was used to detect the effect of ZNF674-AS1 on proliferation of hepatoma cells and detect apoptosis-related gene B cell lymphoma/leukemia-2 associated X protein (bax), B cell lymphoma/leukemia-2 (bcl-2) protein expression level. The apoptosis levels of hepatoma cells in the blank control group and ZNF674-AS1 knockout group were detected by TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometry. At the same time, the expression level of Wnt/β-catenin signaling pathway-related proteins was detected by immunoblotting. Results The expression of ZNF674-AS1 in liver cancer tissues was about 9.6 times higher than that in adjacent tissues (P<0.05). After inhibition of ZNF674-AS1 by siRNA, the proliferation of HepG2 and QGY7701 hepatoma cells was significantly inhibited. The number of clone formation in NC-siRNA group and ZNF674-AS1 siRNA group was 282.13±2.13 vs. 86.00±10.21 for HepG2; 225.52±1.93 vs. 64.00±6.82 for QGY7701 (P<0.05), respectively. In addition, bax expression was significantly up-regulated in both cell lines (for HepG2, 2.35±0.23 vs. 11.35±1.32; for QGY7701, 3.47±1.45 vs. 14.22±1.58), while bcl-2 expression level was inhibited (for HepG2, 12.17±0.39 vs. 4.22±1.92; for QGY7701, 13.52±1.92 vs. 3.89±1.42). TUNEL and flow cytometry results revealed that knockdown of ZNF674-AS1 in HepG2 and QGY7701 hepatoma cells increased the apoptotic rate by 24.56-fold and 14.88-fold, respectively (P<0.05). After knockdown of ZNF674-AS1, the expression of Wnt/β-catenin signaling pathway protein was inhibited in both cell lines (P<0.05). Conclusion Inhibition of ZNF674-AS1 expression in hepatoma cells can inhibit the proliferation of hepatoma cells and promote apoptosis. The mechanism may be related to ZNF674-AS1-mediated Wnt/β-catenin signaling pathway. Key words: Carcinoma, hepatocellular; ZNF674-AS1; Proliferation; Apoptosis; Wnt/β-catenin