The effect of thermal damage on the cell proliferation and invasive metastasis of HCC: an experimental study in vitro / 介入放射学杂志

ABSTRACT

Objective To evaluate the influence of thermal damage on the cell proliferation,invasive metastasis and epithelial-mesenchymal transition of hepatocellular carcinoma (HCC) through experiments in vitro,and to explore the relationship between thermal ablation and the recurrence,metastasis of HCC.Methods The McA-RH7777 HCC cell thermal damage model was established by using external heating method.The effect of thermal damage on the proliferation of HCC cells was detected by Kit-8 assay (CCK-8),and the cell cycle changes were studied by flow cytometry.The effect of thermal damage on the invasion potential of HCC cells was assessed by using Transwell assay.Fluorescence quantitative polymerase chain reaction (RT-PCR) and Western blot were used to evaluate the influence of thermal damage on HCC cell invasion potential,and on the mRNA and protein expression levels of EMT-related molecular markers,including VEGF,MMP-9,Nm23,E-cadherinand vimentin.Results Heating treatment of McA-RH7777HCC cells was performed by putting the cells in 43.5℃ water basin for 30 min.Two to five days after heating treatment the cell proliferative ability was significantly higher than that of control group (P＜0.05).At 48-72hours after heating treatment the proportion of HCC cells in G1 phase was obviously reduced and the proportion of HCC cells in S+G2 phase was significantly increased,the differences were statistically significant (P＜0.05).Compared with the control group,the difference in HCC cell invasion potential determined at 24 h after heating treatment was not significant,while the HCC cell invasion potential determined at 72 h after heating treatment was strikingly increased (22.3±2.46 vs.14.2±l.82,P＜0.001).Real-time PCR and Western blotting results indicated that at 72 h after heating treatment the expression levels of VEGF,MMP-9 and vimentin were significantly increased,while the expression level of E-cadherin was remarkably decreased,the differences were statistically significant (P＜0.05).Conclusion Thermal damage with sub-lethal heating dose can induce McA-RH7777 HCC cell to develop epithelial-mesenchymal transition and to enhance its proliferation and invasive metastasis potential,and HCC cells show higher malignant potential.