17 February 2019 : Laboratory Research
Phosphatase Actin Regulator-1 (PHACTR-1) Knockdown Suppresses Cell Proliferation and Migration and Promotes Cell Apoptosis in the bEnd.3 Mouse Brain Capillary Endothelial Cell Line
Yao Jing1BCDE, Lin Zhang1BC, Zhiming Xu1CDE, Hao Chen1AB, Shiming Ju1ABC, Jun Ding1ABEFG, Yan Guo1CDE*, Hengli Tian1AFGDOI: 10.12659/MSM.912586
Med Sci Monit 2019; 25:1291-1300
Abstract
BACKGROUND: The phosphatase actin regulator-1 (PHACTR-1) gene on chromosome 6 encodes an actin and protein phosphatase 1 (PP1) binding protein, Phactr-1, which is highly expressed in brain tissues. Phactr-1 expression is involved in physiological and pathological cerebral microvascular events. This study aimed to investigate the role of expression of Phactr-1 in a mouse brain capillary endothelial cell line, bEnd.3, by knockdown the PHACTR-1 gene.
MATERIAL AND METHODS: Three bEnd.3 cell groups were studied, CON (normal control cells), NC (control scramble transfected cells), and KD (cells with PHACTR-1 gene knockdown). The PHACTR-1 gene was knocked down using transfection with small hairpin RNA (shRNA). In the three cell groups cell proliferation, migration, and apoptosis were studied by MTT and colony formation assays, transwell and scratch assays, and flow cytometry. The related cell pathways of associated with Phactr-1 knockdown were studied by Western blot.
RESULTS: Phactr-1 knockdown suppressed bEnd.3 cell proliferation and migration, promoted cell apoptosis, and downregulated the expressions of migration-associated proteins, including matrix metalloproteinase (MMP)-2 and MMP-9 and upregulated apoptosis-associated proteins, including Bax, Bcl-2, cleaved caspase-3, and caspase-3.
CONCLUSIONS: Phactr-1 was shown to have a role in the inhibition of endothelial cell proliferation and migration, promoted cell apoptosis, and regulated matrix metalloproteinases and apoptosis-associated proteins. These findings indicate that the expression of the Phactr-1 should be studied further in the cerebral microvasculature, both in vitro and in vivo, regarding its potential as a diagnostic and therapeutic target for cerebral microvascular disease.
Keywords: Acid Phosphatase, Actins, Cell Migration Assays, endothelial cells, Brain, Cell Line, Down-Regulation, Gene Knockdown Techniques, Microfilament Proteins, Phosphoric Monoester Hydrolases, Transfection
1341 25
Editorial
01 November 2023 : Editorial
Editorial: Factors Driving New Variants of SARS-CoV-2, Immune Escape, and Resistance to Antiviral Treatments as the End of the COVID-19 Pandemic is DeclaredDOI: 10.12659/MSM.942960
Med Sci Monit 2023; 29:e942960
In Press
28 Nov 2023 : Clinical Research
Impact of Fluorodeoxyglucose-Positron Emission Tomography/Computed Tomography on Therapeutic Decisions and ...Med Sci Monit In Press; DOI: 10.12659/MSM.942122
28 Nov 2023 : Clinical Research
Long-Term Outcomes of Decompression and Grafting in Acute Pathological Proximal Femur Fractures in Children...Med Sci Monit In Press; DOI: 10.12659/MSM.943031
27 Nov 2023 : Clinical Research
Comparison of Outcomes from Emergency Admissions to a Major Trauma Center in Turkey of 1646 Elderly Patient...Med Sci Monit In Press; DOI: 10.12659/MSM.942916
27 Nov 2023 : Clinical Research
Evaluating Modified Ultrasound-Guided Serratus Anterior Plane Block for Enhanced Postoperative Recovery in ...Med Sci Monit In Press; DOI: 10.12659/MSM.942757
Most Viewed Current Articles
13 Nov 2021 : Clinical Research
Acceptance of COVID-19 Vaccination and Its Associated Factors Among Cancer Patients Attending the Oncology ...DOI :10.12659/MSM.932788
Med Sci Monit 2021; 27:e932788
30 Dec 2021 : Clinical Research
Retrospective Study of Outcomes and Hospitalization Rates of Patients in Italy with a Confirmed Diagnosis o...DOI :10.12659/MSM.935379
Med Sci Monit 2021; 27:e935379
14 Dec 2022 : Clinical Research
Prevalence and Variability of Allergen-Specific Immunoglobulin E in Patients with Elevated Tryptase LevelsDOI :10.12659/MSM.937990
Med Sci Monit 2022; 28:e937990
08 Mar 2022 : Review article
A Review of the Potential Roles of Antioxidant and Anti-Inflammatory Pharmacological Approaches for the Man...DOI :10.12659/MSM.936292
Med Sci Monit 2022; 28:e936292