Expression And Clinical Significance Of HSP90? In Hepatocellular Carcinoma

Objective: 1.To detect the expression of HSP90? in peripheral blood of patients with HCC,analyze the expression characteristics,and compare with AFP to explore the diagnostic efficacy of HSP90? on HCC.2.To detect the expression of HSP90? in HCC and normal liver tissues,analyze the expression characteristics of HSP90? in HCC tissues,and analyze the connection between HSP90? and clinicopathologic features.3.To improve the follow-up information after radical treatment of patients with HCC,and to explore the value of HSP90? in the prognosis evaluation of patients with HCC.4.To simulate the thermal ablation environment of HCC in vivo,to detect the expression level of HSP90? in hepatoma cells under high temperature and hypoxia,and to investigate the changes of HSP90? in hepatoma cells under high temperature and hypoxia,provide a theoretical basis for the study of the mechanism of HSP90? in the recurrence of HCC after thermal ablation.Method: 1.The peripheral blood of 25 patients with first-episode HCC and 20 patients with liver cirrhosis were collected.20 healthy individuals were used as the control group.Enzyme-linked immunosorbent assay was used to detect the expression of serum AFP and HSP90? in patients with HCC,cirrhosis and healthy ones.The diagnostic efficacy of HSP90? on HCC were compared with AFP.2.51 cases of liver tissue specimens with complete medical records and follow-up information were selected,including 30 ones with first-episode HCC and 21 ones with normal liver tissues.Immunohistochemical method was used to detect the expression level of HSP90? in normal liver tissues and HCC tissues,and analyze the the correlation between HSP90? and clinicopathological factors in patients with HCC.3.Improved the follow-up information of 30 patients with HCC after radical treatment,and compared the difference of progression-free survival between high and low expression groups of HSP90?.4.Hepatocellular carcinoma cell line(HepG2)was stimulated by high temperature and hypoxia.Western blot was used to detect the expression level of HSP90? in HepG2 cells and in its condition-induced supernatant.Results: 1.ELISA results showed that HSP90? was significantly higher in the HCC group(139.81�51.98 ng/mL)than in the cirrhosis group(89.26�25.66 ng/mL)and healthy control group(51.67�18.53 ng/mL)(P<0.001;P<0.001);The ROC curve analysis showed that the cutoff value of HSP90? was 92.17 ng/ml,the area under the curve was 0.912,the sensitivity was 92.0%,and the specificity was 82.5%.The upper limit of AFP normal value was 15 ng/ml,and the sensitivity was 56.0%,the specificity was 90.0%.2.The results of IHC showed that HSP90? in HCC was mainly located in tumor cytoplasm,and a small amount was located in the nucleus.The expression level of HSP90? in HCC(mean optical density: 0.24�0.31)was significantly higher than that in liver tissue(mean optical density: 0.17�0.02,P<0.001).Combined with clinicopathological factors,the expression level of HSP90? in HCC was correlated with tumor size and pathological grade(P=0.010;P<0.001).The larger the tumor volume,the lower the degree of differentiation.,the higher expression of HSP90?.3.30 patients with HCC underwent radical treatment,9 patients underwent thermal ablation and 21 underwent surgical resection.Survival analysis showed that HCC patients with high expression of HSP90? had significantly shorter progression-free survival(P=0.006).4.The result of Western blot showed that HSP90? began to increase under heat stress at 45 �C.The secretion of HSP90? began to increase at 6 h after heat stimulation,and the secretion began to peak at 12 h.HSP90? in the cell culture supernatant at 24 h significantly increased compared with the normal control group.There was no significant change in the expression level of HSP90? in cells.The secretion of extracellular HSP90? in hypoxia conditions was increased compared with that in normal oxygen environment,and the secretion of HSP90? was significantly increased after thermal stimulation at 45 �C.Conclusion: 1.HSP90? is elevated in peripheral blood of patients with HCC,and has a high diagnostic efficiency for HCC.It is expected to help AFP to improve the diagnostic sensitivity of HCC and become a new molecular marker of HCC.2.The expression level of HSP90? in HCC tissues is related to tumor size and the degree of differentiation.The larger the tumor volume,the lower the degree of differentiation,the higher the expression level of HSP90?.3.The expression of HSP90? affects the prognosis of patients with HCC.The progression of progression-free survival is significantly shortened in patients with high expression of HSP90?,indicating that HSP90? is important in monitoring HCC progression and prognosis.4.The secretion level of HSP90? was increased under high temperature and hypoxia,but the expression level of intracellular HSP90? was not obvious.Under laboratory conditions,45 �C was suitable temperature for simulated thermal ablation,and thermal stimulation could further increase the secretion of HSP90? under hypoxia.