The Role Of Activated Cdc42-associated Kinase 1 Related Signaling Pathway In Pathogenesis Of Human Hepatocellular Carcinoma

Objective: To clarify the expression of p Tyr284-ACK1 and the its related PTEN/AKT signal pathway molecules in Hepatocellular carcinoma(HCC).To observe the changes of PTEN/AKT/m TOR activation and the biological behavior.Aiming to get potential drug target,and providing a theoretical evidence for the treatment of HCC.Methods: IHC was used to detect the expression of p Tyr284-ACK1, p Ser473-AKT and PTEN protein in HCC tissues, and to analyze its clinical pathological significance.The drugs of AIM-100 and Dasatinib were used to culture the SKHep-1 and Hep G2 cells.The capability of proliferation,migration and invasion were estimated by MTT,Wound health and Transwell methods respectively.Flow cytometry was used to detect the cell cycle distribution and apoptosis.The q PCR and Western blot were used to detect the expression of ACK1, p Tyr284-ACK1,AKT, p Ser473-AKT, m TOR and PTEN, EGFR on m RNA and protein stages.Results:The positive rate of p Tyr284-ACK1 in HCC was higher than the nontumorous adjacent tissues,the high expression was significant in cancer thrombus and metastasis(P<0.05). The high expression of p Ser473-AKT was significant in tumor differentiation degree(P < 0.05). The low expression of PTEN was significant in preoperative AFP, tumor differentiation degrees,capsule invasion,cancer thrombus and recurrence and metastasis(P < 0.05). p Tyr284-ACK1 was positively correlation with p Ser473-AKT(rs=0.336, P < 0.001), and negatively correlation with PTEN(rs=-0.441, P < 0.001). p Ser473-AKT was negatively correlation PTEN(rs=--0.332, P < 0.001). The expression of p Tyr284-ACK1,p Ser473-AKT, PTEN and HCC patients had no significant correlation with tumor free survival period. The median duration of overall survival in patient of low p Tyr284-ACK1 expression were 22.15 months, longer than patients with high expression of 16.79 months(P < 0.05); p Ser473-AKT expression and survival period were not significant correlation; The median duration of overall survival in patient oflow PTEN expression were 18.78 months, shorter than the normal expression in patients with a median survival of 23.88 months(P<0.05).AIM-100 and Dasatinib inhibited the proliferation of SK-Hep-1 and Hep G2 cells in a certain range of concentration.In the Wound health test and Transwell test,the migration and invasion capability of the treated SK-Hep-1(both the AIM-100 and Dasatinib)were significantly lower than the normal,but theresults of Hep G2 cell were not so obviously.The cells were stopped in the G1 phase,and more easily to apoptosis under the treatments of AIM-100 and Dasatinib.In m RNA level,under the treatment of AIM-100,ACK1,AKT and PTEN expressed higher while m TOR and EGFR expressed lower compared with untreated SK-Hep-1;ACK1,AKT,m TOR and EGFR expressed lower,and PTEN expressed higher compared with untreated Hep G2;In Dasatinb group,ACK1 and PTEN expressed higher while the ACK1,m TOR and EGFR expressed lower compaired with untreated SK-Hep-1,EGFR expressed lower and ACK1,AKT,m TOR and EGFR expresses higher compared with untreated Hep G2.In protein level,under the treatment of AIM-100,for the SKHep-1 cell, ACK1, p Tyr284-ACK1, AKT, p Ser473-AKT,EGFR expressed significantly lower and PTEN expressed significantly higher than the untreated one(P < 0.05).For the Hep G2 cells,p Ser473-AKT and PTEN expressed significantly higher(P<0.05). Under the treatment of Dasatinib, for the SK-Hep-1cell,shared the results with AIM-100.For the Hep G2,only p Ser473-AKT and PTEN showed significantly higher than the untreated cells(P<0.05).Conclusion: The high expression of p Tyr284-ACK1 promotes the invasion and metastasis of HCC patients. p Tyr284-ACK1 plays a role in promoting cancer through the PTEN/AKT pathway. p Tyr284-ACK1 positively correlated with p Ser473-AKT and negatively correlated with PTEN. Combined detection of p Tyr284-ACK1 and PTEN protein helps us to predict the recurrence and overall survival of HCC patients.ACK1 could promotes the cell proliferation, invasion and metastasis,and regulate the cell cycle distribution and apoptosis via the EGFR/PTEN/AKT/m TOR pathway.AIM-100 and Dasatinib will reduce the function of ACK1 effectively.