目的 分析BCL2L1基因在卵巢癌(OC)细胞中的恶性生物学行为,并探讨其潜在的调控机制.方法 收集2018年2月至2023年4月15例在西安市人民医院接受手术治疗的OC(浆液性)患者的癌组织及癌旁组织样本.同时,从GSE26712数据库下载了正常样本10例和OC样本185例的数据进行分析.采用OC细胞系(SKOV3、Hey、HO8910、A2780)及人卵巢表面上皮细胞(IOSE80)进行实验.采用qRT-PCR检测肿瘤组织,细胞中B细胞淋巴瘤2类似蛋白 1(B-cell lymphoma 2-like protein 1,BCL2L1)的表达.Western blot 实验检测 BCL2L1 和 Wnt/β-catenin 信号相关蛋白的变化.细胞增殖与毒性检测试剂盒(CCK-8)增殖检测、集落形成能力测试、细胞划痕实验、Transwel l实验、流式细胞计数检测细胞的生长,转移和凋亡能力变化.结果 本研究发现BCL2L1在OC组织及细胞中的表达显著高于正常对照(P＜0.001),并且其表达增强与OC细胞的侵袭、迁移、增殖能力72h正相关(P=0.009、P=0.016、P＜0.001),集落形成能力增强(P=0.002).相反,BCL2L1抑制能显著降低这些能力(侵袭能力P=0.006,迁移能力P=0.011,增殖能力72 h,P＜0.001,集落形成能力P＜0.001)并促进细胞凋亡(P＜0.001).通过基因集富集分析(gene set enrichment analysis,GSEA)与干预实验表明,BCL2L1的作用通过Wnt/β-catenin信号通路实现.结论 BCL2L1在OC组织与细胞中高表达,上调BCL2L1可通过Wnt/β-catenin信号通路促进OC细胞增殖、侵袭和迁移有望成为潜在的治疗靶点.
Exploring the function of BCL2L1 in ovarian cancer development and its regulation through the Wnt/β-catenin signaling pathway
Objective To analyze the malignant biological behaviors of BCL2L1 gene in ovarian cancer(OC)cells and to explore its potential regulatory mechanisms.Methods In this study,between February 2018 and April 2023,cancerous and paraneoplastic tissue samples were collected from 15 patients with OC(plasmacytosis)who underwent surgical treatment at Xi'an People's Hospital.Meanwhile,data from 10 normal samples and 185 OC samples were downloaded from the GSE26712 database for analysis.OC cell lines(SKOV3,Hey,HO8910,A2780)and human ovarian surface epithelial cells(IOSE80)were used for the experiments.Tumor tissues were detected by qRT-PCR,and the expression of B-cell lymphoma 2-like protein 1(BCL2L1)was detected in the cells.Western blot assay was used to detect the changes of BCL2L1 and Wnt/β-catenin signaling-related proteins.Cell proliferation and toxicity assay kit(Cell Counting Kit-8,CCK-8)proliferation assay,colony forming ability test,cell scratch assay,Transwell assay,and flow cytometric counting detected changes in cell growth,metastasis,and apoptotic ability.Results In this study,we found that the expression of BCL2L1 in OC tissues and cells was significantly higher than that of normal control(P＜0.001),and its enhanced expression was positively correlated with the invasive,migratory,and proliferative abilities of OC cells(P=0.009 for invasive ability,P=0.016 for migratory ability,and P＜0.001 for proliferative ability for 72 h),and enhanced colony-forming ability(P=0.002).In contrast,BCL2L1 inhibition sig-nificantly reduced these capacities(invasive capacity P=0.006,migratory capacity P=0.011,proliferative capacity 72 h,P＜0.001,colony forming capacity P＜0.001)and promoted apoptosis(P＜0.001).Gene set enrichment analysis(GSEA)with intervention experiments showed that the effects of BCL2L1 were realized through the Wnt/β-catenin signaling pathway.Conclusion BCL2L1 is highly expressed in OC tissues and cells,and up-regulation of BCL2L1 can promote the proliferation,invasion and migra-tion of OC cells through the Wnt/β-catenin signaling pathway is expected to be a potential therapeutic target.
NETL
NSTL
万方数据
