Tripartite motif containing 54 regulates the proliferation,invasion and metastasis of pancreatic cancer
Objective To investigate the regulatory mechanism of tripartite motif containing 54(TRIM54)in pancreatic cancer.Methods Pancreatic cancer cell lines(ASPC-1,BXPC-3,MIA-PaCa-2,PANC-1,SW1990)and human pancreatic ductal epithelial(HPDE)cell lines were purchased from the American type culture collection(ATCC).Real-time quantitative polymerase chain reaction(RT-PCR)and Western blotting were used to detect the expression of TRIM54 in HPDE cells and pancreatic ductal adeno-carcinoma(PDAC)cells.The expression of TRIM54 was knocked down by plasmid(shRNA)transfection.Cell counting kit-8(CCK-8)assay was used to detect the proliferation of cancer cells.Transwell and wound healing assays were utilized to observe the changes of cell invasion and migration.Independent sample t test was performed for comparison between two groups,while analysis of variance was used for comparison a-mong multiple groups.Results The result of RT-PCR showed that the mRNA expression of TRIM54 in HPDE cells was significantly lower than that in pancreatic cancer cells(AsPC-1,BxPC-3,MIA PaCa-2,PANC-1,SW1990)(1.00±0.02 vs.2.05±0.04 vs.1.37±0.03 vs.1.62±0.02 vs.3.18±0.10 vs.3.59±0.07,F=1 042.04,P＜0.05).Similarly,the result of immunoblotting showed that the protein expression of TRIM54 in HPDE cells was significantly lower than that in pancreatic cancer cells(1.00± 0.02 vs.2.04±0.05 vs.1.18±0.01 vs.2.06±0.02 vs.2.42±0.02 vs.2.90±0.06,F=1 360.92,P＜0.05).The abilities of proliferation,invasion and migration of PANC-1 and SW1990 cells were im-paired after TRIM54 depletion by plasmids transfection.For PANC-1 cells,the N-cadherin level in NC group was higher than that in sh-TRIM54 group(1.00±0.03 vs.0.58±0.03 vs.0.74±0.02,F=233.26,P＜0.05).N-cadherin protein level decreased in sh-TRIM54 group as compared with NC group in SW 1990 cells(0.95±0.02 vs.0.66±0.02 vs.0.77±0.02,F=127.64,P＜0.05).Vimentin in NC group was higher than that in sh-TRIM54 group(1.00±0.03 vs.0.71±0.02 vs.0.79±0.03,F=130.09,P＜0.05),while Vimentin in NC group of SW1990 cells was higher than that in sh-TRIM54 group(0.94±0.02 vs.0.69±0.03 vs.0.66±0.02,F=167.04,P＜0.05).In PANC-1 cells,E-cad-herin expression in NC group was lower than that in sh-TRIM54 group(1.00±0.02 vs.1.08±0.04 vs.1.20±0.03,F=34.21,P＜0.05).The E-cadherin expression in NC group of SW1990 cells was lower than that in sh-TRIM54 group(0.95±0.03 vs.1.36±0.03 vs.1.28±0.03,F=218.35,P＜0.05).The recovery experiment implied that WNT agonists enhanced the proliferation,invasion and migration of PANC-1 and SW1990 cells with TRIM54 knockdown.In addition,the expression of N-cadherin and Vim-entin in sh-TRIM54 group was lower than that in sh-TRIM54+SKL2001 group(N-cadherin:0.67±0.02 vs.0.93±0.01,t=18.02,P＜0.05;Vimentin:0.59±0.02vs.0.91±0.03,t=15.99,P＜0.05).Besides,the expression levels of N-cadherin and Vimentin in sh-TRIM54 group were lower than those in sh-TRIM54+SKL2001 group in SW1990 cells(N-cadherin:0.76±0.01 vs.0.88±0.02,t=10.77,P＜0.05;Vimentin:0.68±0.02 vs.1.07±0.03,t=19.78,P＜0.05).In PANC-1 cells,E-cadherin expression in sh-TRIM54 group was higher than that in sh-TRIM54+SKL2001 group(1.31±0.03 vs.1.15±0.02,t=7.32,P＜0.05).The E-cadherin expression in sh-TRIM54 group was higher than that in sh-TRIM54+SKL2001 group in SW1990 cells(1.20±0.06 vs.1.01±0.06,t=4.20,P＜0.05).Conclusion TRIM54 is highly expressed in pancreatic cancer and promotes the proliferation,invasion and migration of pancreatic cancer cells by regulating the WNT signaling pathway.
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