Pu Haihong, Zhang Qingyuan, Zhao Chunbo, Shi Lei, Wang Yan, Wang Jingxuan, Zhang Minghui
Department of Medical Oncology, Harbin Medical University Cancer Hospital, Haping Road 150 of Nangang District, Harbin, Heilongjiang Province, 150086, China.
Department of Radiation Oncology, Harbin Medical University Cancer Hospital, Harbin, 150086, Heilongjiang Province, China.
Pathol Oncol Res. 2018 Jul;24(3):557-565. doi: 10.1007/s12253-017-0277-4. Epub 2017 Jul 25.
Our study aimed to identify key genes involved in the use of fluvastatin and zoledronate against breast cancer, as well as to investigate the roles of vascular endothelial growth factor A (VEGFA) in the malignant behaviors of breast cancer cells. The expression data GSE33552 was downloaded from Gene Expression Omnibus database, including mocked-, fluvastatin- and zoledronate-treated MDA-MB-231 cells. Differentially expressed genes (DEGs) were identified in fluvastatin- and zoledronate-treated cells using limma package, respectively. Pathway enrichment analysis and protein-protein interaction (PPI) network analysis were then performed. Then we used shRNA specifically targeting VEGFA (shVEGFA) to knock down the expression of VEGFA in MDA-MB-231 cells. Cell viability assay, scratch wound healing assay, Transwell invasion assay and flow cytometry were performed to explore the effects of VEGFA knockdown on the malignant behaviors of breast cancer cells. VEGFA was up-regulated in both fluvastatin- and zoledronate-treated breast cancer cells. Moreover, VEGFA was a hub node in PPI network. In addition, VEGFA was successfully knocked down in MDA-MB-231 cells by shVEGFA. Suppression of VEGFA promoted the migration and invasion of breast cancer MDA-MB-231 cells. Suppression of VEGFA inhibited the apoptosis of MDA-MB-231 cells. Our results indicate that up-regulation of VEGFA may prevent the progression of breast cancer after fluvastatin and zoledronate treatment via inducing cell apoptosis and inhibiting migration and invasion. VEGFA may serve as a potential prognostic indicator for clinical outcome in the management of breast cancer.
我们的研究旨在确定氟伐他汀和唑来膦酸用于治疗乳腺癌所涉及的关键基因，并研究血管内皮生长因子A（VEGFA）在乳腺癌细胞恶性行为中的作用。从基因表达综合数据库下载表达数据GSE33552，包括模拟处理、氟伐他汀处理和唑来膦酸处理的MDA-MB-231细胞。分别使用limma软件包在氟伐他汀和唑来膦酸处理的细胞中鉴定差异表达基因（DEG）。然后进行通路富集分析和蛋白质-蛋白质相互作用（PPI）网络分析。接着我们使用特异性靶向VEGFA的短发夹RNA（shVEGFA）来敲低MDA-MB-231细胞中VEGFA的表达。进行细胞活力测定、划痕伤口愈合测定、Transwell侵袭测定和流式细胞术，以探讨VEGFA敲低对乳腺癌细胞恶性行为的影响。在氟伐他汀和唑来膦酸处理的乳腺癌细胞中VEGFA均上调。此外，VEGFA是PPI网络中的一个枢纽节点。另外，shVEGFA成功敲低了MDA-MB-231细胞中的VEGFA。抑制VEGFA促进了乳腺癌MDA-MB-231细胞的迁移和侵袭。抑制VEGFA抑制了MDA-MB-231细胞的凋亡。我们的结果表明，VEGFA的上调可能通过诱导细胞凋亡以及抑制迁移和侵袭来阻止氟伐他汀和唑来膦酸治疗后乳腺癌的进展。VEGFA可能作为乳腺癌治疗中临床结局的潜在预后指标。
