The expression of nicotinamide N-methyltransferase in thyroid carcinoma and its effect on Wnt/β-catenin signal pathway in TPC-1 cells
Objective Explore to the expression of nicotinamide N-methyltransferase(NNMT)in thyroid carcinoma and its effect on the expression Wnt/β-catenin signal pathway in TPC-1 thyroid carcino-ma cells.Methods Detection of NNMT protein and mRNA expression in thyroid carcinoma and paracan-cerous tissues by Western blotting and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).siRNA NNMT(experimental group)and siRNA NC(control group)were transfected in-to TPC-1 cell by liposome LipofectamineTM 3000.The expression of NNMT protein and mRNA was detected by Western blotting and Real-time PCR.Cell viability was measured by methyl thiazolyl tetrazolium(MTT)assay.Cell apoptosis was detected by flow cytometry.The expression of B-cell lymphoma-2(bcl-2),bcl-2-associated X protein(bax),cleaved poly ADP-ribose polymerase(cleaved PARP),proliferating cell nucle-ar antigen(PCNA),matrix metalloproteinase-2(MMP-2),MMP-9,Wnt1,Wnt 3a and β-catenin was de-tected by Western blotting.Group t test was used to analyze differences between groups.Results The ex-pression of NNMT protein(0.84±0.06 vs.0.30±0.02,t=6.312,P＜0.05)and mRNA(1.87±0.15 vs.1.00±0.10,t=6.456,P＜0.05)in cancer group was higher than that in adjacent cancer group.The expression of NNMT protein(0.38±0.01 vs.0.98±0.07,t=6.034,P＜0.05),mRNA(0.37±0.02 vs.1.00±0.10,t=6.256,P＜0.05),cell viability(0.33±0.02 vs.0.49±0.04,t=5.986,P＜0.05),cell invasion number(35.61±2.34 vs.174.39±10.37,t=6.219,P＜0.05),the expression of bcl-2(0.08±0.00 vs.0.42±0.02,t=5.785,P＜0.05),cleaved PARP(0.07±0.00 vs.0.38±0.01,t=5.752,P＜0.05),PCNA(0.40±0.03 vs.1.25±0.10,t=6.3246,P＜0.05),MMP-2(0.07±0.00 vs.0.83±0.05,t=5.952,P＜0.05),MMP-9(0.06±0.00 vs.0.86±0.02,t=6.872,P＜0.05),Wnt1(0.39±0.01 vs.1.23±0.07,t=5.984,P＜0.05),Wnt3a(0.35±0.00 vs.1.03±0.08,t=5.932,P＜0.05)and β-catenin(0.12±0.00 vs.0.56±0.03,t=5.853,P＜0.05)protein in experimental group was lower that in control group.Cell early apoptotic rate[(18.35±1.24)％vs.(6.32±0.70)％,t=5.872,P＜0.05],late apoptotic rate[(20.44±1.73)％vs.(7.40±0.64)％,t=6.582,P＜0.05],the expression of bax(1.29±0.08 vs.0.53±0.02,t=5.986,P＜0.05)protein in experimental group was higher than that in control group.Conclusion Down-regulation of NNMT expression could induce apoptosis of TPC-1 thyroid cancer cells and inhibit cell invasion,which related to inhibition of Wnt/β-catenin signal pathway.
Nicotinamide N-methyltransferaseThyroid carcinomaWnt/β-catenin signal pathwayApoptosisInvasion
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