人Runt相关转录因子3通过调控上皮-间充质转化抑制涎腺腺样囊性癌转移  

Human Runt related transcription factor 3 suppresses the metastasis of salivary adenoid cystic car- cinoma through transcriptional inhibition of epithelial - mesenchymal transition

作　　者：郑传铭[1] 凌志强[2] 葛明华[1] 

机构地区：[1]浙江省肿瘤医院头颈外科,杭州310022 [2]浙江省肿瘤研究所,杭州310022

出　　处：《中华实验外科杂志》2017年第5期836-839,共4页Chinese Journal of Experimental Surgery

基　　金：浙江省自然科学基金面上项目（LY14H160014）;浙江省卫生高层次创新人才培养工程基金项目（浙卫发[2014]108号）;浙江省新世纪151人才工程重点层次项目（浙人社发[2014]150号）

摘　　要：目的观察过表达人Runt相关转录因子3（RUNX3）对腺样囊性癌细胞上皮-间充质转化（EMT）的影响。方法利用慢病毒感染腺样囊性癌SACC-83、SACC-LM细胞，获得稳定过表达RUNX3的细胞株（过表达RUNX3组），并设置空白慢病毒感染的阴性对照细胞株（阴性对照组）。用Western blot法检测RUNX3蛋白过表达；采用实时定量反转录聚合酶链反应（RT-qPCR）和Western blot检测过表达RUNX3对SACC-83、SACC-LM细胞中E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）、N-钙黏蛋白（N-cadherin）、纤维粘连蛋白（Fibronectin）及Snail基因mRNA和蛋白表达水平的影响；采用Western blot检测过表达RUNX3对SACC-83、SACC-LM细胞基质金属蛋白酶（MMP）-2和MMP-9表达的影响。采用细胞划痕实验检测细胞迁移能力。结果慢病毒感染SACC-83、SACC-LM细胞后，其RUNX3蛋白表达[（0.968±0.035）、（0.956±0.037）]均显著上调，与阴性对照组[（0.425±0.031）、（0.196±0.028）]比较差异有统计学意义（P=0.002、P=0.000）。RT-qPCR和Western blot检测表明，与阴性对照组比较，过表达RUNX3组的Vimentin、Fibronectin、Snail mRNA表达下降34.2%～61.9%、蛋白表达下降31.8%～71.8%；而SACC-83细胞的E-cadherin mRNA和蛋白分别上调2.671、2.237倍，SACC-LM细胞的E-cadherin mRNA和蛋白分别上调3.687、5.454倍。Western blot检测显示，与阴性对照组比较，过表达RUNX3组的MMP-2和MMP-9表达显著下降（P=0. 005、P=0.004）。细胞划痕实验显示，过表达RUNX3细胞的迁移率较低。结论RUNX3可以抑制人腺样囊性癌细胞的EMT，这是其抑制腺样囊性癌细胞侵袭与转移的机制之一。ObjectiveTo investigate the effect of expression of human Runt related transcription factor 3 （RUNX3） on epithelial mesenchymal transition （EMT） in adenoid cystic carcinoma cells.MethodsLentivirus vector was transfected into MDA-MB-231 cells to obtain the RUNX3-overexpressed cell line （RUNX3 overexpression group）, and the cell line with blank lentivirus vector infection served as negative control group. After the wild-type RUNX3 was overexpressed in SACC-83 and SACC-LM cells, RUNX3 protein was detected by Western blotting, and the effect of RUNX3 on the expression of E-cadherin, Vimentin, N-cadherin, Fibronectin and Snail at mRNA and protein levels was investigated using Western blotting and real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR） analysis respectively. Matrix metalloproteinase （MMP）-2 and MMP-9 proteins were also detected by Western blotting. The effect of RUNX3 on the migration of SACC cells was measured by cell scratch assay.ResultsAfter SACC-83 and SACC-LM cells were treated with lentivirus-RUNX3, the expression of RUNX3 protein [（0.968±0.035） and （0.956±0.037）] was significantly up-regulated as compared with negative control group [（0.425±0.031） and （0.196±0.028）] with the difference being statistically significant （P=0.002 and P=0.000）. The mRNA expression levels of Vimentin, Fibronectin and Snail decreased from 34.2% to 61.9%, and the protein expression decreased from 31.8% to 71.8%. However, the expression levels of E-cadherin mRNA and protein in SACC-83 cells were up-regulated 2.671 fold and 2.237 fold respectively. The expression levels of E-cadherin mRNA and protein in SACC-LM cells were up-regulated 3.687 fold and 5.454 fold respectively as compared with negative control group. MMP-2 and MMP-9 protein expression levels were significantly down-regulated by RUNX3 overexpression （P=0. 005 and P=0.004）. The migration of SACC-83 and SACC-LM cells was significantly inhibited by RUNX3 overexpression.ConclusionRUNX3 suppr

关 键 词：涎腺腺样囊性癌 RUNT相关转录因子3 上皮-间充质转化 转移 

分 类 号：R739.8[医药卫生—肿瘤]