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慢病毒载体递送的 BCSG1 siRNA 抑制 MDA-MB-231 细胞的增殖和迁移。

BCSG1 siRNA delivered by lentiviral vector suppressed proliferation and migration of MDA-MB-231 cells.

机构信息

Department of Breast Surgery, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518035, P.R. China.

Biobank, The First Affiliated Hospital of Shenzhen University, Second People's Hospital of Shenzhen, Shenzhen, Guangdong 518035, P.R. China.

出版信息

Int J Mol Med. 2018 Mar;41(3):1659-1664. doi: 10.3892/ijmm.2017.3355. Epub 2017 Dec 29.

Abstract

Breast cancer-specific gene 1 (BCSG1), also referred to as γ-synuclein (SNCG), is highly expressed in human infiltrating breast carcinomas, but not in normal or benign breast tissue. The present study aimed to evaluate the effects of BCSG1 siRNA delivered by lentiviral vector on breast cancer cells and investigate the underlying mechanisms. BCSG1 RNAi lentiviral vector was constructed and transfected into MDA-MB-231 cells. BCSG1 mRNA levels were determined by quantitative polymerase chain reaction analysis. Cell proliferation, migration and apoptosis were evaluated by using the cell counting kit‑8, Transwell assay and flow cytometry, respectively, followed by western blotting to determine the relative levels of AKT, extracellular signal‑regulated kinase (ERK), p-AKT and p-ERK expression. BCSG1 mRNA levels were significantly reduced in MDA-MB‑231 cells following transfection of BCSG1 siRNA delivered by lentiviral vector. Cell migration and proliferation were significantly decreased and the cell cycle was arrested. Western blot analysis indicated that the protein levels of p-AKT and p-ERK were significantly lower in the BCSG1 siRNA-treated groups compared with the control and negative control groups. Therefore, BCSG1 siRNA delivered by lentiviral vector was able to significantly reduce BCSG1 expression, suppress cell migration and proliferation, possibly through the reduction of the protein levels of p-AKT and p-ERK.

摘要

乳腺癌特异基因 1(BCSG1),也称为γ-突触核蛋白(SNCG),在人浸润性乳腺癌中高度表达,但在正常或良性乳腺组织中不表达。本研究旨在评估慢病毒载体递送的 BCSG1siRNA 对乳腺癌细胞的影响,并探讨其潜在机制。构建 BCSG1RNAi 慢病毒载体并转染 MDA-MB-231 细胞。采用实时定量聚合酶链反应分析检测 BCSG1mRNA 水平。采用细胞计数试剂盒-8、Transwell 检测和流式细胞术分别评估细胞增殖、迁移和凋亡,然后采用 Western blot 检测 AKT、细胞外信号调节激酶(ERK)、p-AKT 和 p-ERK 表达的相对水平。转染慢病毒载体递送的 BCSG1siRNA 后,MDA-MB-231 细胞中 BCSG1mRNA 水平显著降低。细胞迁移和增殖明显减少,细胞周期停滞。Western blot 分析表明,BCSG1siRNA 处理组的 p-AKT 和 p-ERK 蛋白水平明显低于对照组和阴性对照组。因此,慢病毒载体递送的 BCSG1siRNA 能够显著降低 BCSG1 表达,抑制细胞迁移和增殖,可能通过降低 p-AKT 和 p-ERK 的蛋白水平。

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