Mechanism of dendritic-cell specific 7 transmembrane protein promoting proliferation,migration,and invasion of thyroid papillary carcinoma cells
Objective To investigate the effect and mechanism of dendritic-cell specific 7 trans-membrane protein(DCSTAMP)on the proliferation,migration,and invasion of thyroid papillary carcinoma cells.Methods The expression of DCSTAMP in thyroid papillary carcinoma was analyzed in GEPIA data-base.The expression of DCSTAMP between thyroid papillary carcinoma cell lines KTC-1,FTC-133 and thyroid normal cell line Nthy-ori 3-1 was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting.The knockdown and overexpression plasmids and their controls were transfected respectively into KTC-1 and FTC-133 cells to construct DCSTAMP overexpression and knockdown cell models.In the knockdown group,the effect of DCSTAMP gene knockdown on migration and invasion was detected by cell scratch and Transwell test.Western blotting was used to verify the effect of knockdown on the expression of NCAD,phosphorylated protein kinase B(p-Akt)and total protein ki-nase B(t-Akt)proteins.In the overexpression group,the effect of overexpression of DCSTAMP gene on proliferation was detected using the cell counting kit-8(CCK-8)and the EdU cell proliferation test kit(DAB).Western blotting was used to verify the effect of overexpression on proliferation cell nuclear antigen(Ki-67)protein and mitogen-activated protein kinase(MAPK)pathway.The mean of the two samples was compared by paired t test analysis.Results Both RT-qPCR and Western blotting results showed that the expression of DCSTAMP gene in KTC-1 and FTC-133 was significantly higher than that in Nthy ori 3-1[RT-qPCR(KTC-1:t=4.901,P＜0.05;FTC-133:t=37.460,P＜0.01);Western bloting(KTC-1:t=9.628,P＜0.01;FTC-133:t=55.580,P＜0.01)].When the DCSTAMP gene was overexpressed in thy-roid cancer cells KTC-1 and FTC-133,the rate of cell proliferation was increased(KTC-1 on the day 3:t=6.198,P＜0.01;FTC-133 on the day 3:t=33.640,P＜0.0005),and the proliferation ability was in-creased(KTC-1:t=65.220,P＜0.01;FTC-133:t=49.320,P＜0.01).The expression of Ki-67 and MAPK was increased(P＜0.01).The MAPK pathway subfamily extracellular signal-regulated kinase(ERK)showed varying degrees of elevation(KTC-1:t=68.650,P＜0.01;FTC-133:t=34.000,P＜0.01).After knockdown of DCSTAMP gene in KTC-1 and FTC-133,the cell migration rate slowed down(KTC-1:t=21.020,P＜0.01;FTC-133:t=10.450,P＜0.01).The invasion ability decreased(KTC-1:t=14.940,P＜0.01;FTC-133:t=4.603,P＜0.01).The p-Akt expression decreased(KTC-1:t=21.980,P＜0.01;FTC-133:t=35.900,P＜0.01).Conclusion The increased expression of DCSTAMP gene in thyroid papillary carcinoma cell lines KTC-1 and FTC-133 can promote tumor cell proliferation,mi-gration,and invasion through the activation of MAPK pathway,and Akt phosphorylation.
Thyroid papillary carcinomaDendrites express specific 7 transmembrane proteinsMitogen-activated protein kinase pathwayProtein kinase B phosphorylation
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