The Inhibitation Effect Of Dichloroacetic Acid On Prolif Eration,Invasion And Migration In Hunman Renal Cell Line A489

Objectives: Renal cell carcinoma(Rcc)is one of the most common types of renal parenchymal malignancies,and renal tubular epithelial cells are its cell source.The etiology of kidney cancer is complicated.Currently,smoking,drugs,viruses,and heavy metal cadmium are known.Among them,smoking is one of the known independent risk factors.This study was aimed to investigate the effects of dichloroacetic acid(DCA)on proliferation,invasion and migration of human renal cell line A498,and to analyze its mechanism.Methods: Human renal cancer cell line A498 was cultured and divided into Negative control(NC)group,Positive control(PC)group,DCA low dose,medium dose and high dose groups.3 DCA dose groups were given 5,10,20 mmol/L DCA respectively,NC group was given isovolumetric sterile saline,PC group was given 5-fluorouracil 5 mg/L for 48 hours.MTT assay was used to detect the inhibition rate of proliferation after 24 h and 48 h.Transwell cell test was used to detect the invasive ability.Cell scratch test was used to detect migration ability.RT-q PCR was used to detect the expression of Snail,Slug,E-cadherin,N-cadherin and Vimentin genes.Western blotting(WB)was used to detect the expression of these proteins.Results: The inhibition rates of proliferation in NC group at 48 h and 24 h were similar(P > 0.05).The inhibition rates of proliferation in the other four groups were significantly increased(P < 0.05).Compared with each other at 24 h and 48 h,the inhibition rates in DCA high-dose group were higher than that in DCA medium-dosegroup/PC group,DCA low-dose group and NC group.Except that in DCA medium-dose group and PC group,the difference was similar(P > 0.05).There were significant differences in invasive activity and mobility of each group(P < 0.05).There were significant differences between each two groups(P < 0.05),except DCA medium dose group and PC low dose group,which were similar to PC group(P > 0.05).The expressions of Snail,Slug,E-cadherin,N-cadherin and Vimentin in each group were significantly different(P < 0.05).The expressions of Snail,Slug,N-cadherin and Vimentin mRNA and proteins in DCA high-dose group < DCA medium-dose group /PC < DCA low-dose group < NC group,except DCA medium-dose group and PC group were similar(P > 0.05),there were significant differences between the remaining two groups(P < 0.05).The expressions of E-cadherin mRNA and protein in each two groups was compared with that in DCA high-dose group > DCA medium-dose group / PC group > DCA low-dose group > NC group.Except DCA medium-dose group and PC group were similar(P > 0.05),there were significant differences between the other two groups(P < 0.05).Conclusion: DCA can inhibit the proliferation,invasion and migration of human renal cell line A498 in a dose-dependent manner.The effect of high-dose DCA is better than that of 5-fluorouracil.It is speculated that it may be related to down-regulation of Snail,Slug,N-cadherin,Vimentin and up-regulation of E-cadherin.