The Effect Of Chemotactic Factor CXCL1 Secreted By Hepatic Stellate Cell On Hepatic Carcinoma’s Epithelial-msenchymal Transition

Objective: To explore the effect of hepatic stellate cells on hepatocellular carcinoma(HCC) cells’ epithelial-msenchymal transition and underlying mechanism. Methods: HCC cells(Hep G2, SMMC-7721) were co-cultured with conditional medium of HSC or HSC themselves. Cell migration and invasion were detected by methods of cell wound healing and Transwell chamber assays. Product of CXCL1 in HSC and in HSC-conditional medium(HSC-CM), the expression of CXCR2(CXCL1 receptor——C-X-C-chemokine receptor 2) in HCC cells, and p-PI3 K, p-AKT, p-GSK-3β and Snail in conditional cultivated HCC cells were detected by Western blot. The change of epithelial markers, E-cadherin, mesenchymal markers, N-cadherin and Vimentin in HCC cells were detected by laser scanning confocal microscopy(LSCM) and Western blot. Results: HSC produced much chemokines CXCL1, while HCC cells(Hep G2, SMMC-7721) highly expressed CXCL1 receptor CXCR2. Moreover, their morphology changed, adhesion ability decreased, migration and invasion ability enhanced, P <0.05, compared with the control group. The expression of epithelial markers, E-cadherin, were downregulated and mesenchymal markers, N-cadherin and Vimentin, were up-regulated in conditional cultivated HCC cells. Furthermore, the phosphorylation level of the important members of PI3K/AKT signal pathway, PI3 K and AKT were up-regulated, as well as levels of p-GSK-3β and transcription factor Snail under conditional cultivation, P <0.05, compared with the control group. In contrast, the expression of epithelial markers, E-cadherin, were up-regulated, but mesenchymal markers, N-cadherin and Vimentin, were down-regulated, and intracellular p-PI3 K, p-AKT, p-GSK-3β and Snail were down-regulated, when the conditional cultivated HCC cells treated by CXCR2 inhibitor(SB265610), P <0.05, compared with the group that before treated by inhibitor. Conclusion: Hepatic stellate cells promoted HCC cells migration and invasion. Hepatic stellate cells induced epithelial-mesenchymal transition of HCC through activating PI3 K / AKT/ p-GSK-3β/Snail signaling pathway by CXCL1/CXCR2 axis.