The Impact Of Thyroid Disrupting Chemicals P,p’-dde And Malathion On Protein Expression In Human Thyroid Cells Nthy-ori-3-1

Objective:The thyroid is the largest endocrine gland of human,which plays a key role in maintaining hormonal balance and regulating the function of body’s tissues and organs properly.Many surveys show that the incidence of thyroid diseases are rising in our country.As thyroid disrupting chemicals,p,p’-DDE and Malathion can interfere with thyroid hormone secretion,disrupt the normal thyroid function.In this study,human thyroid cells Nthy-ori-3-1 were used to find the changes of protein expression excluding cytotoxic effect of p,p’-DDE and Malathion to explain the disrupting mechanism.Methods:Single standard Agilent expression profiling microarray was used to screen the differentially expressed genes of the cells.MTT method was applied to study the effects of p,p’-DDE and Malathion on the proliferation of Nthy-ori-3-1 cells.The cells were dealt with the dose of p,p’-DDE and Malathion in 0μg/mL,0.5μg/mL,1μg/mL,2μg/mL in 24 hours.Four genes were selected to detect the protein expression by Western-blot technique,then GEL PRO,Graph PAD were used to analyze the results of protein expression.Results:In the group of Malathion,there are 73 genes showed differences in microarray,including 31 up-regulated genes and 42 down-regulated genes.In the group ofp p.p’-DDE,there are 33 genes showed differences in microarray,including 20 up-regulated genes and 13 down-regulated genes.MTT assay showed that,p,p’-DDE and Malathion had no significant changes on proliferation of cells within the dose of 0μg/mL,0.5μg/mL,1μg/mL,2μg/mL.The relative grey level of DIRAS1 and APOL3 significantly increased in group of 1μg/mL and 2pg/mL compared with those in control group;the protein expression increased,and there were statistical significance(p<0.05).The relative grey level of LHX4 and DIS3L significantly decreased in group of 1μg/mL and 2μg/mL compared with those in control group;the protein expression decreased,and there was statistical significance(p<0.05).Conclusion:DIRAS1 and APOL3 were selected to detect the protein expression in the group of Malathion,which are related in intracellular signal transduction in up-regulated gene.The protein expression of DIRAS1 and APOL3 are increased,which seems to be associated with RAS mutation and the activity change of NF-κB pathway.LHX4 and DIS3L were selected to detect the protein expression in the group of p,p’-DDE,which are related in apoptosis and proliferation in down-regulated gene.The protein expression of LHX4 and DIS3L are decreased,which seems to be associated with Anti-pituitary function and cell proliferation.