The Antitumor Effect And Mechanism Of Chinese Herbal P.nemoralis A.DC.extract Zinc Pyrithione On Renal Clear Carcinoma Cells

Objectives:To study the antitumor effect of Zinc Pyrithione combined with zinc ion on human renal carcinoma cell line.To investigate the mechanism of the antitumor activity of Zinc Pyrithione in human renal clear carcinoma cells.Methods:Cell viability of ZPT plus 50?M ZnCl₂ on 786-O and 769-P cell lines was detected with MTS method.The average inhibitory concentration 50 values(IC50 values)were determined after48-hour treatment.The dose-course and time-course effect of ZPT plus 50?M ZnCl₂ in two cells lines was measured with MTS methods.The colony formation ratio and apoptosis of ZPT plus50?M ZnCl₂ on two cells lines were determined with colony formation assay and flow cytometry.The distribution of free zinc ion and lysosome in 786-O cell was probed with the relative fluorescent probes and observed with fluorescent microscope.The changes of intracellular lysosomal permeability were detected by acridine orange(AO)fluorescence staining.The NF?B transcription activity was measured with the luciferase activity of NF?B reporter plasmid.Results:ZPT plus 50?M ZnCl₂ can inhibit the proliferation of 786-O and 769-P cell lines with the IC50 values 501.8n M and 694.7n M respectively after 48-hour treatment.Proliferation inhibition was a time-and dose-dependent process.The antibacteria rate of 3.2?M ZPT plus 50?M ZnCl₂ is 20.23%and 24.75%in 786-O and769-P cells respectively after 48-hour treatment,43.55%and38.16%in two cell lines after 72-hour treatment with 500n M ZPT for 786-O and 700n M for 769-O in the presence of 50?M Zn Cl_2.ZPT plus 50?M ZnCl₂ inhibited the colony formation in two cell lines(DMSO vs ZnCl₂ vs ZPT+ZnCl₂ :100.00%vs 98.40%vs 40.24%for 786-O cells,P<0.01;100.00%vs 104.37%vs 45.24%for 769-P cells,P<0.01).The apoptosis ratio was enhanced in 786-O cell line after 12-hour treatment of 500n M ZPT+50?M ZnCl₂ (Apoptosis ratio:1.31%,1.85%?3.69%and 25.15%in the groups of DMSO,ZnCl₂ ,ZPT and ZPT+ZnCl₂ ,P<0.01).Pyrithione targeted zinc to lysosome in 786-O cells and lysosomal integrity was disrupted as shown by redistribution of Acridine Orange.Pyrithione plus zinc decrease the NF?B transcription activity in 786-O cells.Conclusions:1.The traditional Chinese medicine extract Anselutra contains zinc ions,which can inhibit the proliferation of 786-O and769-P cells;When combined with zinc ion,the inhibition effect was stronger in a dose-dependent and time-dependent manner.Inhibitory effect of 786-O and 769-P on apoptosis of renal clear carcinoma cells.2.The combination of ZPT and zinc ion can inhibit the proliferation of renal cancer cells,and the inhibition of NF?B activity and the increase of lysosomal membrane permeability are the possible mechanisms.