Effect Of Curcumin Derivative C086 On Senescent And Quiescent Hepatoma Cells

Hepatocellular carcinoma(HCC)is one of the most common malignancies in the world.Tumor metastasis and recurrence is the main cause of death after treatment.However,the process of metastasis is very complex,involving the adhesion,migration and invasion of tumor cells,the secretion of MMPase and the degradation of extracellular matrix.Therefore,it is very important to overcome the recurrence of liver cancer.Previous studies have shown that C086 has antihepatocellular carcinoma activity via inhibiting HSP90 in HCC cells,which can be an effective therapy for HCC to prolong the survival rate for liver cancer patients.Objective:To investigate the antitumor activity and possible mechanism of C086 against static and senescent hepatocellular carcinoma cells in vitro.Method:MTT method was used to detect the inhibition of C086 on ADR-induced senescent HepG2 and QGY7703 cells,low-glucose and low serum-induced quiescent liver cancer cells.(2)β-galactosidase staining was used to detect the effect of C086 on senescent HepG2 and QGY7703 cells.(3)Apoptosis was measured using Annexin V-APC/PI staining in senescent HepG2 and QGY7703 cells and quiescent liver cancer cells.(4)Scratch test method was used to detect the effect of C086 on the motility of senescent HepG2 cells.(5)Transwell chamber method was used to detect the effect of C086 on the migration and invasion of HepG2 cells.(6)q PCR was used to detect the effect of C086 on the expression levels of P16,P21 and IL6 in senescent HepG2 cells.(7)Western Blot analysis was used to measure the expression of inflammation-related SASP.(8)DCFH fluorescent probe method was used to measure the level of reactive oxygen species in HepG2 cells.(9)Chemiluminescence method was used to measure the level of ATP in quiescent HepG2 cells.Result:(1)Adriamycin(ADR)induced senescence in HepG2 cells at a concentration of0.125 μg/m L.Senescence was confirmed using β-galactosidase staining,β-galactosidase fluorescence substrate C12 FDG,and the upregu Lation of expressions of P16 and P21.(2)When senescent HCC cells treated with C086,the expression ofβ-galactosidase positive cells significantly decreased,the cell size decreased,and the expression of senescent markers γ-H2 AX,P16,P21 also decreased.(3)Annexin V-APC/PI staining showed that C086 promoted apoptosis of senescent cells.(4)SCM induced by senescent supernatant showed slow proliferation and EMT characteristics,which was reversed by C086 treatment.(5)Supernatants containing IL-6 from the culture of senescent cells can also induce cell senescence.(6)QGY7703cells showed the same characteristics as HepG2 cells.(7)The expression levels of AMPK,NF-κB,m TOR proteins in senescent HCC cells induced by ADR and senescent supernatant were decreased in the same manner.(8)C086 inhibited the growth of HepG2 cells in quiescent phase induced under hypoxia and low glucose conditions,and promote its apoptosis.(9)C086 increased the production of ROS and decreased the production of ATP in the quiescent phase of HepG2 cells induced by hypoxia and hypoglycemia.Conclusion:(1)Curcumin derivative C086 can inhibit the growth and apoptosis of hepatocellular carcinoma cells HepG2 and QGY7703,and the mechanism may be related to the decrease of AMPK-NF-k B/m TOR-NF-k B.(2)Curcumin derivative C086 can inhibit the growth and apoptosis of HepG2 cells in quiescent stage.