In-vitro study of epithelial-mesenchymal transition mediated by HBX protein and M2 macrophages in hepatocellular carcinoma cells / 中华普通外科杂志

ABSTRACT

Objective To explore the synergetic effect of HBX protein and M2 macrophages in inflammatory microenvironment on invasion and metastasis of hepatocellular carcinoma cells.Methods Hep3B cells were infected with recombinant lentivirus carrying HBx gene,following co-culture with THP-1 original M2 macrophages.The cells were divided into six groups:two infected groups (Hep3B +and Hep3B + + M2),four non-infected groups (Hep3B-,Hep3B-+ LV5,Hep3B-+ M2,Hep3B-+LV5 + M2).Western blot (WB) was used to assess the expression changes of E-cadherin and N-cadherin,markers of epithelial-mesenchymal transition (EMT).The cellular location of EMT markers was observed by immunofluorescence confocal microscopy.Transwell assay was used to evaluate the invasion ability of Hep3B cells.Results HBX protein overexpressed in Hep3B cells by lentivirus infection.After 72 h co-culture with M2 macrophages,WB results showed that E-cadherin descreased significantly in Hep3B+ (0.42 ±0.11) when compared with Hep3B-(1.00 ±0.18) (t =4.762,P ＜0.05),while N-cadherin was significantly higher in Hep3B + (2.85 ± 0.44) than in Hep3B-(1.00 ± 0.17) (t =4.762,P ＜ 0.05).M2macrophages decreased E-cadherin expression in Hep3 B + + M2 (0.1 ± 0.13) compared with Hep3 B + (t =3.255,P ＜0.05),while N-cadherin expression increased in Hep3B+ + M2 (4.18 ± 0.52) (t=10.009,P ＜ 0.05).Non-Infected groups didn't change the markers of E-cadherin and N-cadherin.It was suggested that invasion ability of Hep3B increased by HBx overexpression.Conclusions HBX protein and M2 macrophages synergetically mediated the invasion and metastasis of hepatocellular carcinoma cells by EMT.