Cancer Biology Laboratory, Faculty of Life Science & Biotechnology, South Asian University, Akbar Bhawan, Chankyapuri, New Delhi, India.
PLoS One. 2019 May 13;14(5):e0211658. doi: 10.1371/journal.pone.0211658. eCollection 2019.
Earlier, we have identified PTOV1 as a novel interactome of PIN1 in PC-3 cells. This study aims to explore the functional similarity and the common role of both genes in breast cancer cell proliferation.
CTG, crystal violet assay, clonogenic assay, wound healing assay, cell cycle analysis, Hoechst staining and ROS measurement were performed to assess cell viability, colony forming potential, cell cycle arrest, nuclear condensation and ROS production after knocking down of PTOV1 and PIN1 by siRNAs in MDA-MB-231 and MCF-7 cells. CO-IP, qPCR and western blot were performedto study interaction, transcriptional and translational regulation of both genes.
Knockdown of PTOV1 and PIN1 inhibited the cell proliferation, colony formation, migration, cell cycle, and induced nuclear condensation as well as ROS production. Interaction of PTOV1 and PIN1 was validated by Co-IP in MDA-MB-231 cells. Genes involved in cell proliferation, migration, cell cycle, and apoptosis were regulated by PIN1 and PTOV1. PTOV1 knockdown inhibited Bcl-2, Bcl-xL and inducedBAX, LC3 and Beclin-1expression. Overexpression of PIN1 increased the expression of PTOV1. Knockdown of both genes inhibited the expression of cyclin D1, c-Myc, and β-catenin.
PTOV1 and PIN1 interact and exert oncogenic role in MDA-MB-231 cells by sharing the similar expression profile at transcriptional and translational level which can be a promising hub for therapeutic target.
我们之前发现 PTOV1 是 PC-3 细胞中 PIN1 的一个新的相互作用体。本研究旨在探索这两个基因在乳腺癌细胞增殖中的功能相似性和共同作用。
通过 siRNA 敲低 MDA-MB-231 和 MCF-7 细胞中的 PTOV1 和 PIN1，采用 CTG、结晶紫分析、集落形成分析、划痕愈合分析、细胞周期分析、Hoechst 染色和 ROS 测量来评估细胞活力、集落形成潜力、细胞周期阻滞、核浓缩和 ROS 产生。通过 CO-IP、qPCR 和 Western blot 来研究两个基因的相互作用、转录和翻译调控。
敲低 PTOV1 和 PIN1 抑制了 MDA-MB-231 细胞的增殖、集落形成、迁移、细胞周期，并诱导核浓缩和 ROS 产生。在 MDA-MB-231 细胞中通过 CO-IP 验证了 PTOV1 和 PIN1 的相互作用。细胞增殖、迁移、细胞周期和凋亡相关基因受 PIN1 和 PTOV1 调控。PTOV1 敲低抑制了 Bcl-2、Bcl-xL 并诱导了 BAX、LC3 和 Beclin-1 的表达。过表达 PIN1 增加了 PTOV1 的表达。敲低这两个基因抑制了 cyclin D1、c-Myc 和 β-catenin 的表达。
PTOV1 和 PIN1 通过在转录和翻译水平上具有相似的表达谱相互作用，并在 MDA-MB-231 细胞中发挥致癌作用，这可能是一个有前途的治疗靶点。
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