骨诱导因子通过NF2/Hippo信号通路抑制甲状腺乳头状癌细胞增殖和侵袭  

Osteoglycin regulates proliferation and invasion of papillary thyroid carcinoma cells via NF2/Hippo signaling pathway

作　　者：樊东 齐玉娟 赵华栋 FAN Dong;QI Yujuan;ZHAO Huadong(Department of General Surgery,Second Affiliated Hospital of Air Force Medical University,Xi′an 710032,China)

机构地区：[1]空军军医大学第二附属医院普通外科,西安710032

出　　处：《山西医科大学学报》2024年第3期269-279,共11页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81372859)。

摘　　要：目的探究骨诱导因子(osteoglycin,OGN)在甲状腺乳头状癌(PTC)中的作用和调控机制。方法利用基因表达谱交互分析(GEPIA)数据库分析甲状腺肿瘤组织和正常组织中OGN表达的差异及患者无病生存期。采用实时荧光定量PCR和蛋白免疫印迹分析我院的52例PTC肿瘤组织和相应的癌旁正常组织中OGN表达。实时荧光定量PCR和蛋白免疫印迹分析PTC细胞系(K1,TPC-1,IHH-4,KTC-1和BCPAP)和正常甲状腺细胞系(Nthy-ori3-1)中OGN表达水平。使用对照质粒和pcDNA3.1-OGN质粒分别转染TPC-1和KTC-1细胞,分为:control组、pc-NC组和pc-OGN组;随后,将si-RNA-si-RNA-NF2或si-RNA-LATS1共转染至pcDNA3.1-OGN组TPC-1细胞中,分为:pc-NC组、pc-OGN组、pc-OGN+si-NF2组和pc-OGN+si-LATS1组。采用CCK-8、流式细胞术、Transwell和划痕实验分别检测细胞增殖、凋亡、侵袭和迁移水平;蛋白免疫印迹法检测上皮间充质转化相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)以及NF2和Hippo信号通路关键蛋白Yes关联蛋白1(YAP1)、哺乳动物STE20样蛋白激酶1(MST1)和大肿瘤抑制因子1(LATS1)的蛋白表达。结果GEPIA数据库分析结果显示,与正常组织相比,肿瘤组织中OGN表达水平减少(P<0.01),低水平的OGN表达与较短的无病生存期显著正相关。同时,临床患者样本中肿瘤组织OGN表达水平也较正常组织有明显减少(P<0.01)。与Nthy-ori3-1细胞比较,5种PTC细胞系中OGN mRNA和蛋白表达均有不同程度的减少(P<0.01)。与pc-NC组比较,pc-OGN组TPC-1和KTC-1细胞活力、侵袭和迁移能力减弱(P<0.01),上皮间充质转化相关蛋白N-cadherin和Vimentin表达减少(P<0.01),而细胞凋亡率增加(P<0.01),E-cadherin蛋白表达增加(P<0.01)。与pc-NC组比较,pc-OGN组细胞中NF2表达水平升高(P<0.01),Hippo通路YAP1磷酸化水平减少,MST1和LAST1磷酸化水平增加(P<0.01)。与pc-OGN组细胞相比,pc-OGN+si-NF2组和pc-OGN+si-LATS1组细胞凋亡率显著Objective To explore the role and its regulatory mechanism of osteoglycin(OGN)in papillary thyroid carcinoma(PTC).Methods The expression of OGN and the disease-free survival time in thyroid cancer patients were analyzed using gene expression profile interactive analysis(GEPIA)database.OGN expression in 52 cases of PTC tissues and adjacent normal tissues from our hospital was detected by real-time quantitative PCR and Western blotting.OGN expression in PTC cell lines(K1,TPC-1,IHH-4,KTC-1 and BCPAP)and normal thyroid cell line(Nthy-ori3-1)was also analyzed by real-time quantitative PCR and Western blotting.TPC-1 and KTC-1 cells were divided into control group,pc-NC group(transfected with pcDNA3.1 empty plasmid)and pc-OGN group(transfected with pcDNA3.1-OGN plasmid).Then,TPC-1 cells were co-transfected with pcDNA3.1-OGN plasmid and si-RNA-NF2 or pcDNA3.1-OGN plasmid and si-RNA-LATS1,respectively,namely pc-NC group,pc-OGN group,pc-OGN+si-NF2 group and pc-OGN+si-LATS1 group.Cell proliferation,apoptosis,invasion and migration were detected by CCK-8,flow cytometry,Transwell and wound healing assays,respectively.The protein expressions of epithelial mesenchymal transition related proteins(E-cadherin,N-cadherin,Vimentin),NF2 and key proteins in Hippo signaling[Yes associated protein 1(YAP1),mammalian STE20-like protein kinase(MST1)and large tumor suppressor 1(LATS1)]were detected by Western blotting.Results GEPIA database analysis showed that OGN expression in tumor tissues was significantly lower than that in normal tissues(P<0.01),and low OGN expression level was closely correlated with shorter disease-free survival time.OGN expression in tumor tissues was significantly lower than that in adjacent normal tissues(P<0.01).Compared with Nthy-ori3-1 cells,OGN mRNA and protein expression levels in PTC cell lines were decreased to varying degrees(P<0.01).Compared with pc-NC group,cell proliferation,invasion and migration ability of TPC-1 and KTC-1 cells were significantly inhibited in pc-OGN group(P<0.01),and N-cadherin and V

关 键 词：骨诱导因子 甲状腺乳头状癌 细胞活力 凋亡 侵袭 迁移 NF2/Hippo通路 

分 类 号：R736.1[医药卫生—肿瘤]