Influence Of Hcpt On Proliferation Of Human Adrenal Cortex Carcinoma Cell Line SW-13 And Wnt/β-catenin Signaling Pathway

Objective To observe the changes of proliferation and apoptosis after treatment with hydroxycamptothecine(RCPT),an anticancer drug,on human adrenoeortical carcinoma cell line SW-13 in vitro,and to investigate on molecular level effect of hydroxycamptothecine on extracellular inhibitors of the wnt/β-catenin pathway(DKK-1)and the key protein β-catenin in wnt/β-catenin signaling pathway,to discuss its possible molecular mechanism.Method human adrenoeortical carcinoma cells SW-13 cultured in vitro were exposed to avariety of concentrations(0.25、0.5、1.0、2.0、4.0ug/mL)of HCPT for different time(24h、48h、72h),the inhibitory effect of HCPT on cell SW-13 proliferation weas measured by CKK8 assay.Calculate the IC50 of HCPT and select the best duration of action;Select IC50 as follow-up experiments concentration and experiment were divided into two groups--the HCPT intervention group and control group.The cell apoptosis was measured by flow cytometry when treated with HCPT.Quantitative PCR tests was used to analyze the transcription of DKK-1 and β-catenin after treatment of HCPT for 48 hours;The expression of DKK-1 and β-catenin was evaluated in SW-13 cell lines using immunocytochemistry.Results The HCPT of different concentrations could significantly inhibited the SW-13 cell proliferation(P<0.05)and showed a time and concentration dependent manner;Flow cytometry show higher apoptosis rate of SW-13 cell in the HCPT intervention group than the control group(P<0.05).Quantitative PCR tests and immunocytochemistry showed that the expression of DKK-1 was higher in the HCPT intervention group than the control group(P<0.05).P-catenin expression did not change significantly(P>0.05).Conclusion The results have showed that HCPT can significantly inhibit the growth’ and proliferation of adrenalcortical carcinoma SW-13 cells,and induce apoptosis.HCPT could promotes the expression of DKK-1 genes.However,increased expression of DKK-1 genes did not cause the change of theβ-catenin mRNA levels and cellular localization.These results indicate HCPT-induced apoptosis in SW-13 cells might be regulated through influencing the expressions of DKK-1 genes.DKK1 have no effect on the canonicalβ-catenin dependent pathway in SW-13 cells and may mediated theβ-catenin-independent pathways in anti-tumor effect.