The Effects Of Knocking Down GATAD2A Gene On Proliferation?cell Cycle And Apoptosis In Thyroid Carcinoma

ObjectivesTo analyze human thyroid carcinoma cell's proliferation,cell cycle and apoptosis after the GATAD2 A gene silencing.Thus may offer a reference for the clinical diagnosis and prognosis if thyroid carcinoma,and may provide a novel target in gene's diagnosis and therapy.Methods 1.Constructing lentivirus-delivered shorthairpin(shRNA),Transfecting 293 T cells,measuring the transfection rate and virus drops;2.Lentivirus-delivered shorthairpin(shRNA)were transfected into thyroid carcinoma cells,then the mRNA and protein of GATAD2 A were measured by RT-PCR ? Western Blot to detect the GATAD2A's silencing effect.3.MTT and flow cytometry were applied to detect the proliferation and cell cycle in thyroid carcinoma cells influenced by shGATAD2A(S1,S2),shCon.4.Western blot were applied to detect the apoptotic index and the expression of caspases-3,PARP in thyroid carcinoma cells influenced by shGATAD2A(S1,S2)contrasting to shCon.5.Statistical analysis.All the experiments were repeated three times.The SPSS 17.0 was used to anal-yze the results with t-test or one-way ANOVA.The results were considered to be statistically significant at Pvalue <0.05.Results 1.Three recombined plasmids: shGATAD2A(S1,S2),shCon were constructed successfully.After transfe-cting 293 T cells,the transfection rate: > 90%,virus titer were: shCo : 7.0E+06 Tu/ml;shGATAD2A(S1): 4.0E+06 Tu/ml;shGATAD2A(S2): 7.0E+06 Tu/ml 2.Lentivirus-delivered shorthairpin(shRNA)were transfected into thyroid carcinoma cells successfully,the transfection rate: >80%.The expression of mRNA and protein of GATAD2 A was obviously lower in thyroid carcinoma cells transfected by shGATAD2A(S1,S2)than those by shCon.3.The growth rates of thyroid carcinoma cells transfected by shGATAD2A(S1,S2)are obviously slower than shCon group.The percentage of cells in G0/G1 phase was significantly decreased,while the cells in G2/M phase were remarkably increased in shGATAD2 A infected GATAD2 A.4.The apoptotic index and the expression of caspases-3 protein and PARP in thyroid carcinoma cells transfected by shGATAD2 A are obviously higher than those transfected by shCon?Conclusions 1.We have constructed and Packaged the Lentivirus-delivered shorthairpin(shRNA).2.The mRNA and protein of GATAD2 A in thyroid carcinoma cells be knocked down effectively by Lentivirus-delivered shRNA.3.Knocking down the GATAD2 A gene can promote thyroid carcinoma cells proliferation and lead to thyroid carcinoma cell cycle arrest.4.Knocking down the GATAD2 A gene can promote apoptotic of thyroid carcinoma cells.