Inhibiting the Progression of Human Retinoblastoma Cell by Downregulation of MMP-2MMP-9 Using Short Hairpin RNAs (shRNAs) In Vitro
Joint Authors
Meng, Nana
Zhao, Zhi
Shi, Chunhe
Xia, Leizhou
Source
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-05-14
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
Objective.
To investigate the effect of downregulated matrix metalloproteinases (MMPs) gene on the proliferation, apoptosis, cell cycle, migration, and invasion of human retinoblastoma (RB) cell line in vitro.
Methods.
Small hairpin RNA (shRNA) targeting MMP-2/MMP-9 was designed and transfected into WER1-Rb-1 cells.
48 hours after transfection, qRT-PCR and western blot technique were used to investigate the inhibitory effect of MMP-2 and MMP-9 shRNAs.
Cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
Cell cycle arrest was detected using a flow cytometer while apoptosis was tested with Annexin V/PI kit.
Transwell chamber assay was performed to detect the migration and invasion ability of the WER1-Rb-1 cells.
Results.
After transfection of MMP-2/MMP-9 shRNA, there was a significant decrease in the expressions of both mRNA and protein in the shRNA groups compared with the negative and vector controls.
The results of MTT assay suggested that the cell viability was significantly decreased in shRNA groups (p<0.05).
Cell apoptosis also increased significantly in shRNA groups compared with the negative and vector controls (p<0.05).
The flow cytometer analysis proved that the proportion of the G1 phase increased and the proportion of the G0 phase reduced significantly by the transfection of MMP-2/MMP-9 shRNA (p<0.05).
The migration and invasion ability were also significantly decreased in the groups of MMP-2/MMP-9 shRNA (p<0.05).
Conclusions.
Cell viability, migration, and invasion ability of RB cells are inhibited, and apoptosis is induced after downregulation of MMP-2/MMP-9 through RNA interference.
MMP-2 and MMP-9 may be potential targets in the gene therapy of RB.
American Psychological Association (APA)
Meng, Nana& Zhao, Zhi& Shi, Chunhe& Xia, Leizhou. 2020. Inhibiting the Progression of Human Retinoblastoma Cell by Downregulation of MMP-2MMP-9 Using Short Hairpin RNAs (shRNAs) In Vitro. Journal of Ophthalmology،Vol. 2020, no. 2020, pp.1-10.
https://search.emarefa.net/detail/BIM-1189437
Modern Language Association (MLA)
Meng, Nana…[et al.]. Inhibiting the Progression of Human Retinoblastoma Cell by Downregulation of MMP-2MMP-9 Using Short Hairpin RNAs (shRNAs) In Vitro. Journal of Ophthalmology No. 2020 (2020), pp.1-10.
https://search.emarefa.net/detail/BIM-1189437
American Medical Association (AMA)
Meng, Nana& Zhao, Zhi& Shi, Chunhe& Xia, Leizhou. Inhibiting the Progression of Human Retinoblastoma Cell by Downregulation of MMP-2MMP-9 Using Short Hairpin RNAs (shRNAs) In Vitro. Journal of Ophthalmology. 2020. Vol. 2020, no. 2020, pp.1-10.
https://search.emarefa.net/detail/BIM-1189437
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1189437