首页|DARS2调控EGFR促进膀胱癌的EMT、迁移及侵袭

DARS2调控EGFR促进膀胱癌的EMT、迁移及侵袭

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目的 探讨线粒体天冬氨酰-tRNA合成酶2(DARS2)对膀胱癌细胞上皮间质转化(EMT)、迁移及侵袭的作用及其机制.方法 利用TIMER和GEPIA数据库分析DARS2在膀胱尿路上皮癌(BLCA)中的表达及其与临床病理分期的关系.RT-qPCR检测人正常膀胱上皮细胞SV-HUC-1,膀胱癌细胞5637、T24和TCCSUP中DARS2 mRNA水平.TCCSUP细胞分为:control 组(未转染)、si-NC 组(转染阴性对照 siRNA)、si-DARS2-1 组(胞转染 DARS2 siRNA-1)、si-DARS2 组(转染 DARS2 siRNA-2)、si-DARS2+Vector 组(转染 DARS2 siRNA-2+空载质粒)和 si-DARS2+OE-EGFR 组(转染 DARS2 siRNA-2+EGFR 过表达质粒).Western blot 检测 DARS2,EMT 标志蛋白 E-cadherin、N-cadherin、Vimentin 及 EGFR 的表达水平.Trans well 侵袭和细胞划痕实验分别检测细胞侵袭能力和迁移能力.结果 DARS2 mRNA水平在BLCA中显著上调,并与病理分期正相关(F=3.57,P=0.028 9).与永生化人正常膀胱上皮SV-HUC-1细胞相比,DARS2在5637、T24和TCCSUP膀胱癌细胞中表达水平显著升高(P<0.05).与 si-NC 组相比,si-DARS2 组中 E-cadherin 表达水平显著上调(P<0.05),N-cadherin、Vimentin 和 EGFR蛋白表达水平显著下调(P<0.05);与si-NC组相比,si-DARS2组细胞侵袭数目和细胞迁移率显著降低(P<0.01).与si-DARS2+Vector组相比,si-DARS2+OE-EGFR组E-cadherin蛋白表达水平显著下降(P<0.05),N-cadherin和Vimentin蛋白表达水平显著升高(P<0.05),细胞侵袭数目和细胞迁移率显著增加(P<0.01).结论 敲低DARS2通过下调EGFR表达抑制膀胱癌的EMT、侵袭及迁移.
DARS2 promotes EMT,migration and invasion of bladder cancer through regulating EGFR
Objective To investigate the role of mitochondrial aspartyl-TRNA synthetase 2(DARS2)in epithelial mesenchymal trans-formation(EMT),migration and invasion of bladder cancer cells,and explore its underlying mechanism.Methods DARS2 expression in bladder urothelial carcinoma(BLCA)and its relationship with clinicopathological stage were analyzed using TIMER and GEPIA data-bases.RT-qPCR was used to measure the DARS2 mRNA expression in human normal bladder epithelial cells SV-HUC-1 and bladder cancer cells 5637,T24 and TCCSUP.TCCSUP cells were divided into control group(no transfection),si-NC group(transfected with negative control siRNA),si-DARS2-1 group(transfected with DARS2 siRNA-1),si-DARS2 group(transfected with DARS2 siRNA-2),si-DARS2+Vector group(co-transfected with DARS2 siRNA-2 and empty plasmid)and si-DARS2+OE-EGFR group(co-transfected with DARS2 siRNA-2 and EGFR overexpressing plasmid).Western blot was used to detect the expression of EMT markers(E-cadherin,N-cadherin,Vimentin)and EGFR.Transwell invasion assay and cell scratch assay were conducted to detect cell invasion and migration ability,respectively.Results DARS2 mRNA level was significantly higher in BLCA tissue than in adjacent normal tissue,and it was positively correlated with the pathological stage of the disease(F=3.57,P=0.028 9).Compared with immortalized human normal bladder epithelial SV-HUC-1 cells,DARS2 expression was significantly increased in bladder cancer cells 5637,T24 and TCCSUP(P<0.05).Compared to si-NC group,the protein expression levels of N-cadherin,Vimentin,and EGFR were significantly downregulated in si-DARS2 group(P<0.05),while the protein expression of E-cadherin was significantly upregulated(P<0.05).The number of cell invasion and the cell mobility were significantly lower in si-DARS2 group than in si-NC group(P<0.01).Compared with si-DARS2+Vector group,the expression of E-cadherin was significantly decreased in si-DARS2+OE-EGFR group(P<0.05),while the expressions of N-cadherin and Vimentin were significantly increased(P<0.05).Compared with si-DARS2+Vector group,the number of cell invasion and the cell mobility in si-DARS2+OE-EGFR group were significantly increased(P<0.01).Conclusion DARS2 knockdown can inhibit bladder cancer EMT,invasion and migration via downregulating EGFR expression.

DARS2bladder urothelial carcinomaEGFRepithelial-mesenchymal transitionmigrationinvasion

李华锋、张鸿毅、肖克兵、杨辉、赵刚刚

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西安医学院第一附属医院泌尿外科,西安 710077

DARS2 膀胱癌 EGFR EMT 迁移 侵袭

2024

山西医科大学学报
山西医科大学

山西医科大学学报

CSTPCD
影响因子:0.931
ISSN:1007-6611
年,卷(期):2024.55(3)
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