Lipofection of single guide rna targeting mmp8 decreases proliferation and migration in lung adenocarcinoma cells
Maradiaga O.D.H. Mok P.L. Sivapragasam G. Samrot A.V. Ali Khan M.S. Farhana A. Alzahrani B. Tong J. Karuppiah T. Joseph N.M.S. Subbiah S.K.
July 2021MDPI AG
Medicina (Lithuania)
2021#57Issue 7
Background and Objectives: Matrix metalloproteinases (MMP) have been implicated as major determinants of tumour growth and metastasis, which are considered two of the main hallmarks of cancer. The interaction of MMP8 and other signalling molecules within and adjacent tumoral tissues, including immune cells, are rather elusive, particularly of adenocarcinoma cell type. In this study, we aimed to investigate the role of MMP8 in non-small cell lung cancer proliferation and invasiveness potential. Materials and Methods: We individually lipofected with two different single guide RNA (sgRNAs) that specifically targeted on MMP8, with CRISPR-Cas 9 protein into the cells. Results: Our results clearly indicated that the lipofection of these complexes could lead to reduced ability of A549 cells to survive and proliferate to form colonies. In addition, when compared to non-transfected cells, the experimental cell groups receiving sgRNAs demonstrated relatively decreased migration rate, hence, wider wound gaps in scratch assay. The quantitative real time-polymerase chain reaction (qRT-PCR) demonstrated significant reduction in the MAP-K, survivin and PI3-K gene expression. MMP8 might have protective roles over tumour growth and spread in our body. Conclusions: The delivery of sgRNAs targeting on the MMP8 gene could induce tumour cell death and arrest cell migratory activity.
CRISPR-Cas9 , Epithelial to mesenchymal transition (EMT) , Lipofection , Lung cancer , Matrix metalloproteinase 8 (MMP8) , Single guide RNA
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Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, Serdang, 43400, Malaysia
Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, P.O. Box 2014, Sakaka, Saudi Arabia
Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, 43400, Malaysia
Genetics and Regenerative Medicine Research Group, Universiti Putra Malaysia, UPM, Serdang, 43400, Malaysia
Institute of Bioscience, Universiti Putra Malaysia, Serdang, 43400, Malaysia
School of Bioscience, Faculty of Medicine, Bioscience and Nursing, MAHSA University, Jenjarom, 42610, Malaysia
Department of Biomedical Sciences, School of Medicine, Nazarbayev University, Nur-Sultan, 010000, Kazakhstan
Department of Pharmacology, Hamidiye International Faculty of Medicine, University of Health Sciences, Mekteb-I, Tibbiye-I Sahane (Hamidiye) Complex Selimiye Mahallesi, Tibbiye Caddesi #38, Istanbul, 34668, Turkey
Department of Biotechnology, Bharath Institute of Higher Education and Research, 173, Agaram Main Rd, Selaiyur, Chennai, 600073, India
Department of Medical Microbiology and Parasitology
Department of Clinical Laboratory Sciences
Department of Biomedical Science
Genetics and Regenerative Medicine Research Group
Institute of Bioscience
School of Bioscience
Department of Biomedical Sciences
Department of Pharmacology
Department of Biotechnology
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