title: A novel and selective inhibitor of PKC ζ potently inhibits human breast cancer metastasis in vitro and in mice.
abstract: Cell motility and chemotaxis play pivotal roles in the process of tumor development and metastasis. Protein kinase C ζ (PKC ζ) mediates epidermal growth factor (EGF)-stimulated chemotactic signaling pathway through regulating cytoskeleton rearrangement and cell adhesion. The purpose of this study was to develop anti-PKC ζ therapeutics for breast cancer metastasis. In this study, a novel and high-efficient PKC ζ inhibitor named PKCZI195.17 was screened out through a substrate-specific strategy. MTT assay was used to determine the cell viability of human breast cancer MDA-MB-231, MDA-MB-435, and MCF-7 cells while under PKCZI195.17 treatment. Wound-healing, chemotaxis, and Matrigel invasion assays were performed to detect the effects of PKCZI195.17 on breast cancer cells migration and invasion. Adhesion, actin polymerization, and Western blotting were performed to detect the effects of PKCZI195.17 on cells adhesion and actin polymerization, and explore the downsteam signaling mechanisms involved in PKC ζ inhibition. MDA-MB-231 xenograft was used to measure the in vivo anti-metastasis efficacy of PKCZI195.17. The compound PKCZI195.17 selectively inhibited PKC ζ kinase activity since it failed to inhibit PKC α, PKC β, PKC δ, PKC η, AKT2, as well as FGFR2 activity. PKCZI195.17 significantly impaired spontaneous migration, chemotaxis, and invasion of human breast cancer MDA-MB-231, MDA-MB-435, and MCF-7 cells, while PKCZI195.17 did not obviously inhibited cells viability. PKCZI195.17 also inhibited cells adhesion and actin polymerization through attenuating the phosphorylations of integrin β1, LIMK, and cofilin, which might be the downstream effectors of PKC ζ-mediated chemotaxis in MDA-MB-231 cells. Furthermore, PKCZI195.17 suppressed the breast cancer metastasis and increased the survival time of breast tumor-bearing mice. In summary, PKCZI195.17 was a PKC ζ-specific inhibitor which dampened cancer cell migration and metastasis and may serve as a novel therapeutic drug for breast cancer metastasis.
authors.affiliation: Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China. wujing_821013@sina.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China.
authors.forename: Shuye
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China.
authors.forename: Zhijuan
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Clinical Laboratory, Tianjin Third Central Hospital, No. 83 Jintang Road, Hedong District, Tianjin, 300170, China.
authors.forename: Yaqiong
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Research Center of Basic Medical Science, Tianjin Medical University, No. 22 Qixiangtai Road, Heping District, Tianjin, 300070, China. yangrui_cn@hotmail.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Yang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects; D005260:Female; D006801:Humans; D051379:Mice; D009362:Neoplasm Metastasis / Q000473:pathology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D000099221:Protein Kinase C zeta
chemical_list: D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D000099221:Protein Kinase C zeta
keywords: Breast cancer; Chemotaxis; Inhibitor; Metastasis; PKC ζ