The effect of targeted regulation of LATS2 by LncRNA BCAR4 on proliferation, migration and apoptosis of HCC cells.

OBJECTIVE: This study explored and analyzed the effects of targeted regulation of LATS2 by LncRNA BCAR4 on the proliferation, migration and apoptosis of hepatocellular carcinoma (HCC). METHOD: We detected the expression of LncRNA, BCAR4 and LATS2 mRNA in liver hepatocellular carcinoma HepG2 cells and normal hepatocellular cells LO2 by RT-PCR. HepG2 cells were divided into BCAR4-siRNA, NC-siRNA and control groups. We detected the targeted regulation of LncRNA BCAR4 on LATS2 by luciferase gene assay, and measured the proliferation, migration and apoptosis of cells in each group by RT-PCR, MTT, Transwell and flow cytometry, respectively. RESULTS: The relative expression of LncRNA BCAR4 in HepG2 cells was critically higher than that in LO2 cells (P<0.05), while LATS2 mRNA in HepG2 cells was significantly less than that in LO2 cells (P<0.05). Compared with NC siRNA group, the content of luciferase in BCAR4 siRNA group was much higher (P<0.05); The relative expression of LncRNA BCAR4 in BCAR4 siRNA group decreased dramatically than that in NC-siRNA and control group (P<0.05), and the relative expression of LATS2 mRNA increased remarkably than that in NC-siRNA group and control group (P<0.05). The OD value of BCAR4 siRNA group was dramatically higher than that of NC-siRNA group and control group after 48 h and 72 h culture (P<0.05). The quantity of invaded cells in BCAR4 siRNA group was markedly less than that in NC-SIRNA group and control group (P<0.05). Cell apoptosis rate in BCAR4-siRNA group was significantly higher than that of NC-siRNA group and control group (P<0.05). CONCLUSION: LncRNA BCAR4 can regulate the LATS2 expression, and inhibiting the expression of LncRNA BCAR4 can inhibit proliferation, invasion of HepG2 cells and induce its apoptosis, which finding provides a certain reference for the targeted therapy of liver cancer.

Expression of Wnt3a in hepatocellular carcinoma and its effects on cell cycle and metastasis.

Invasion and metastasis are the primary causes of mortality from hepatocellular carcinoma (HCC). Effective inhibition against participants in the tumourigenesis and metastasis process is critical for treatment of HCC. Wnt3a is involved in the development and metastasis of many malignant tumours. However, the specific mechanisms of Wnt3a-mediated cell proliferation, invasion and metastasis in HCC remain unclear. In this study, we found that Wnt3a and its target gene c­Myc showed higher expression in tumour tissues than normal liver tissues in HCC patients; 71.8% of the cases studied had high Wnt3a and c­Myc expression levels (n=32); Wnt3a expression positively correlated with its target genes MMP­7 and c­Myc. Intriguingly, the expression of Wnt3a, MMP­7 and c­Myc is significantly correlated with Notch3 and Hes1 expression. In vitro experiments showed that Wnt3a was highly expressed in MHcc97H and SK­Hep­1 cells. Therefore, Wnt3a expression was silenced with siRNA, and then, MTT, flow cytometry, wound healing and Transwell assays were performed to analyse cell proliferation, cycle, migration and invasion. The results demonstrated that downregulation of Wnt3a expression inhibited cell viability and induced G0/G1 cell cycle arrest via decreased expression of cyclin D1 and c­Myc and increased expression of p21 and p27. In addition, deletion of Wnt3a significantly inhibited migration and invasion by downregulating MMP­2/-7/-9 expression via the MAPK (p38, ERK1/2 and JNK) pathway. In conclusion, our data show that Wnt3a is involved in HCC development. Wnt3a may be an effective target for treatment of HCC.