摘要
目的基于Oncomine数据库分析细胞周期蛋白D1(Cyclin D1)基因在卵巢癌中的表达水平及沉默其表达对癌细胞生物学行为的影响。方法利用Oncomine数据库中关于卵巢癌组织中Cyclin D1基因的相关信息,分析Cyclin D1基因在不同肿瘤中的表达情况以及在卵巢癌中的表达情况。使用在线数据库Kaplan-Meier Plotter分析Cyclin D1表达水平与卵巢癌预后的关系。使用qRT-PCR检测各卵巢癌细胞株(SKOV3、CAOV3、A2780、ES2)及正常卵巢上皮细胞中Cyclin D1的表达。取对数生长期卵巢癌细胞SKOV3,将细胞分为空白对照组、阴性对照组(NC siRNA组)和Cyclin D1基因沉默组(Cyclin D1 siRNA组);使用Western blotting检测各组Cyclin D1蛋白表达;分别使用MTT法、细胞划痕实验和Transwell实验检测各组SKOV3细胞的增殖、迁移和侵袭能力。结果通过挖掘分析Oncomine数据库涉及的Cyclin D1基因发现,与正常卵巢组织比较,在所有差异表达基因中,Cyclin D1基因的中位数值排名为202.0,P<0.001。与Cyclin D1低表达的卵巢癌患者比较,Cyclin D1高表达患者的总生存期和无瘤进展生存期均较短,差异有统计学意义(P<0.01)。与正常卵巢上皮细胞比较,Cyclin D1基因在各卵巢癌细胞(ES2、CAOV3、SKOV3、A2780)中的表达明显升高,差异有统计学意义(P<0.05);与SKOV3细胞株比较,Cyclin D1基因在CAOV3、A2780、ES2细胞株中的表达明显降低,差异有统计学意义(P<0.05)。Cyclin D1 siRNA组细胞中Cyclin D1的表达量、增殖能力、迁移面积、穿过聚碳酸酯膜的癌细胞数量均明显低于空白对照组和NC siRNA组,差异有统计学意义(P<0.05)。结论Cyclin D1基因在卵巢癌组织中高表达,该基因可能参与卵巢癌的发生发展和转移,并与患者预后密切相关。
Objective Study on the role and mechanism analysis of Cyclin D1 gene in the ovarian cancer based on Oncomine database.Methods The expression of Cyclin D1 gene in different tumors and ovarian cancer was analyzed by using the relevant information of Cyclin D1 gene in Oncomine database.The relationship between the expression of Cyclin D1 and the prognosis of ovarian cancer was analyzed by online database Kaplan-Meier Plotter.The expression of Cyclin D1 in ovarian cancer cell lines(SKOV3,CAOV3,A2780,ES2)and normal ovarian epithelial cells was detected by qRT-PCR.Ovarian cancer SKOV3 cells in logarithmic phase were divided into blank control group,negative control group(NC siRNA)and Cyclin D1 gene silencing group(Cyclin D1 siRNA group).The expression of Cyclin D1 protein in each group was detected by Western blotting,and the proliferation,migration and invasion of SKOV3 cells were detected by MTT method,cell scratch test and Transwell,respectively.Results By mining and analyzing the Cyclin D1 genes involved in Oncomine database,it was found that compared with normal ovarian tissues,the median value of Cyclin D1 gene was 202.0,P<0.001.compared with ovarian cancer patients with low expression of Cyclin D1,the overall survival time and progression-free survival time of patients with high expression of Cyclin D1 were significantly lower than those of patients with low expression of Cyclin D1(all P<0.01).Compared with normal ovarian epithelial cells,the expression of Cyclin D1 gene was significantly higher in ES2,CAOV3,SKOV3 and A2780 ovarian cancer cells(all P<0.05).The expression of Cyclin D1 gene in SKOV3 cell lines was significantly lower than that in ES2,CAOV3 and A2780 cell lines(all P<0.05).The expression of Cyclin D1,proliferation,migration area and the number of cancer cells passing through polycarbonate membrane in Cyclin D1 siRNA group were significantly lower than those in blank control group and NC siRNA group(P<0.05).Conclusions Cyclin D1 gene is highly expressed in ovarian cancer.This gene may be involved in the occurrence,development and metastasis of ovarian cancer and is closely related to the prognosis of patients.
作者
刘铃铃
刘振贤
程虹
LIU Lingling;LIU Zhenxian;CHENG Hong(Department of Obstetrics and Gynecology,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine,Haikou 570208, China)
出处
《中国肿瘤外科杂志》
CAS
2022年第2期182-189,共8页
Chinese Journal of Surgical Oncology
