摘要
目的探讨人滋养层细胞表面抗原2(Trop2)在卵巢癌侵袭和转移中的作用及其分子机制。方法通过对Cancer Cell Line Encyclopedia(CCLE)和The Cancer Genome Atlas(TCGA)数据库进行数据挖掘,分析Trop2表达的临床意义。采用Western blot法检测卵巢癌细胞系A3O、A1780和SKOV3中Trop2蛋白的表达。采用Trop2-shRNA构建SKOV3-shRNA细胞模型,以定量逆转录聚合酶链反应检测SKOV3-shRNA和SKOV3-NC组细胞中SKOV3 mRNA的表达,以细胞计数盒8法检测SKOV3-shRNA组和SKOV3-NC组细胞的增殖,以流式细胞仪检测SKOV3-shRNA和SKOV3-NC组细胞的细胞周期和凋亡,以Transwell实验检测SKOV3-shRNA和SKOV3-NC组细胞的侵袭和迁移,以Western blot法检测SKOV3-shRNA和SKOV3-NC组细胞中AKT、p-AKT、β-catenin、caspase3、bcl-2、E-cadherin和vimentin蛋白的表达。结果 Trop2 mRNA在卵巢癌中高表达,其表达高低与肿瘤分期有关,也与患者预后有关。与A3O细胞比较,A1780和SKOV3细胞高表达Trop2蛋白(P<0.05)。SKOV3-NC组和SKOV3-shRNA组中Trop2 mRNA的相对表达量分别为1.18±0.24和0.42±0.08,差异有统计学意义(P<0.05)。SKOV3-NC组细胞的活力明显高于SKOV3-shRNA组(P<0.05)。SKOV3-NC和SKOV3-shRNA组G_(0)/G_(1)期细胞的比例分别为(38.67±4.22)%和(60.24±8.17)%,SKOV3-shRNA组细胞阻滞于G_(0)/G_(1)期(P<0.05)。SKOV3-shRNA组细胞的凋亡率为(26.32±1.81)%,明显高于SKOV3-NC组[(6.54±1.32)%,P<0.05]。SKOV3-shRNA组和SKOV3-NC组SKOV3细胞的迁移细胞数分别为(1 255.83±108.44)个/视野和(1 679.71±213.92)个/视野,侵袭细胞数分别为(242.49±52.09)个/视野和(473.54±73.11)个/视野,与SKOV3-NC组比较,SKOV3-shRNA组的迁移和侵袭细胞数均明显减少(均P<0.05)。SKOV3-shRNA细胞中p-AKT、bcl-2、vimentin、β-catenin表达下调,caspase3、E-cadherin表达上调,总AKT没有明显变化。结论 Trop2的表达与卵巢癌分期和预后有关,Trop2可通过激活AKT/β-catenin信号通路促进卵巢癌细胞增殖和转移,沉默Trop2的�
Objective To explore the role and molecular mechanism of trophoblastic cell surface antigen 2(Trop2)in the invasion and migration of ovarian cancer.Methods Through the data mining of Cancer Cell Line Encyclopedia and TCGA database,the clinical significance of Trop2 expression was analyzed.Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O,A1780 and SKOV3.SKOV3 cells were used to construct Trop2-short hairpin RNA(shRNA)cell model.Quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR)was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells.Cell counting kit-8(CCK8)was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells.Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells.Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells.Western blot was used to detect the protein expressions of AKT,p-AKT,β-catenin,caspase3,bcl-2,E-cadherin and vimentin.Results Trop2 mRNA highly expressed in ovarian cancer,and was related to the tumor stage and patient survival.Compared with A3O cells,Trop2 overexpressed in A1780 and SKOV3 cells(P<0.05).The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08,with statistically significant difference(P<0.05).The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group(P<0.05).The proportion of G_(0)/G_(1) cells in SKOV3-NC and SKOV3-shRNA groups were(38.67±4.22)%and(60.24±8.17)%,respectively.G_(0)/G_(1) arrest was observed in SKOV3-shRNA cells(P<0.05).The apoptosis rate of SKOV3-shRNA group was(26.32±1.81)%,significantly higher than(6.54±1.32)%of SKOV3-NC group(P<0.05).The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1255.83±108.44 and 1679.71±213.92,while the number of invading cells were 242.49±52.09 and 473.54±73.11,respectively.Compared with t
作者
刘明博
史许锋
刘侃
宋婉玉
王莉
武海英
吴广银
Liu Mingbo;Shi Xufeng;Liu Kan;Song Wanyu;Wang Li;Wu Haiying;Wu Guangyin(Department of Cancer Center,Henan Provincial People′s Hospital,Zhengzhou 450003,China;Department of Obstetrics,Henan Provincial People′s Hospital,Zhengzhou 450003,China)
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2021年第3期299-305,共7页
Chinese Journal of Oncology
关键词
卵巢肿瘤
侵袭
转移
预后
Ovarian neoplasms
Invasion
Metastasis
Prognosis
