PFKFB3促进鼻咽癌细胞增殖和转移的作用及潜在机制研究OA

AIM:To investigate the effects of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3(PFKFB3)on the proliferation and metastasis of nasopharyngeal carcinoma(NPC)cells,and to analyze its protein expres-sion profile and associated signaling pathways.METHODS:PFKFB3 overexpression(oe-)and knockdown(sh-)lentivi-ruses were constructed,and their effects were analyzed using real-time cellular analysis(RTCA)and wound-healing and Transwell assays.Subcutaneous xenograft and footpad-inguinal lymph node metastasis models were established in nude mice to evaluate the effects of PFKFB3 on tumor growth and metastasis in vivo.Label-free data-independent acquisition-based quantitative proteomics was used to identify differentially expressed proteins(DEPs)and enriched pathways.Key DEPs were verified using Western blot and immunohistochemistry(IHC).RESULTS:Overexpression of PFKFB3 signifi-cantly enhanced NPC cell proliferation,migration,and invasion in vitro,together with down-regulation of E-cadherin and up-regulation of X-linked inhibitor of apoptosis protein(XIAP),proliferating cell nuclear antigen(PCNA),survivin,N-cadherin,vimentin,occludin,and annexin A1(ANXA1)(P<0.05 or P<0.01).The opposite effects were observed in PFKFB3-knockdown cells.In vivo,PFKFB3 overexpression promoted tumor growth and up-regulated XIAP,PCNA,N-cadherin,vimentin,and ANXA1 in tumor tissues,whereas knockdown inhibited tumor growth and metastasis,down-regu-lated the above proteins and survivin,and up-regulated E-cadherin(P<0.05 or P<0.01).Proteomic analysis identified 407 DEPs in PFKFB3-knockdown cells,and enrichment analyses indicated involvement of these proteins in the Notch,Hippo,Rap1,and transforming growth factor-β(TGF-β)signaling pathways.Specifically,the levels of pro-oncogenic proteins such as recombination signal binding protein for immunoglobulin κJ region(RBPJ),plasminogen activator inhibi-tor-1(PAI-1/SERPINE1),proto-oncogene tyrosine-protein kinase SRC,and profilin 2 were decreased,whereas those of tumor-suppressive proteins such as TGF-β receptor 2(TGFBR2)and cyclin-dependent kinase 4 inhibitor B(CDKN2B)were increased.These changes in expression were verified using IHC in xenograft tissues(P<0.05 or P<0.01).CON-CLUSION:PFKFB3 functions as an oncogene in NPC,with expression levels significantly correlated with proliferation,migration,invasion,and in vivo metastasis of NPC cells.Aberrant expression of PFKFB3 may promote malignant progres-sion of NPC by regulating key proteins in signaling pathways,such as the Notch,Rap1,Hippo and TGF-β pathways.