Lipofection of Single Guide RNA Targeting <i>MMP8</i> Decreases Proliferation and Migration in Lung Adenocarcinoma Cells

• Main Authors: Oscar David Hernandez Maradiaga, Pooi Ling Mok, Gothai Sivapragasam, Antony V. Samrot, Mohammed Safwan Ali Khan, Aisha Farhana, Badr Alzahrani, Jiabei Tong, Thilakavathy Karuppiah, Narcisse M. S. Joseph, Suresh Kumar Subbiah
• Format: Article
• Language: English
• Published: MDPI AG 2021-07-01
• Series: Medicina
• Subjects: CRISPR-Cas9; single guide RNA; lipofection; lung cancer; epithelial to mesenchymal transition (EMT); matrix metalloproteinase 8 (<i>MMP8</i>)
• Online Access: https://www.mdpi.com/1648-9144/57/7/710

<i>Background and Objectives</i>: 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 <i>MMP8</i> 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 <i>MMP8</i> in non-small cell lung cancer proliferation and invasiveness potential. <i>Materials and Methods</i>: We individually lipofected with two different single guide RNA (sgRNAs) that specifically targeted on <i>MMP8</i>, with CRISPR-Cas 9 protein into the cells. <i>Results</i>: 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 <i>MAP-K</i>, <i>survivin</i> and <i>PI3-K</i> gene expression. <i>MMP8</i> might have protective roles over tumour growth and spread in our body. <i>Conclusions</i>: The delivery of sgRNAs targeting on the <i>MMP8</i> gene could induce tumour cell death and arrest cell migratory activity.