Molecular Cloning, Expression, and Sub-Cellular Localization of MGLL in NSCLC Cell Lines
Youssra Boustany1, Abdelilah Laraqui1, Mohammed Oukabli2, Yassine Sekhsokh1, Bouchra Belkadi3, Steven Gray41Research and Biosafety Laboratory, Mohamed V Military Teaching Hospital, Faculty of Medicine and Pharmacy, Mohammed V University in Rabat, Morocco., 2Anatomopathology Laboratory, Mohamed V Military Teaching Hospital, Faculty of Medicine and Pharmacy, Mohammed V University in Rabat, Morocco., 3Microbiology and Molecular Biology Team, Faculty of Sciences, Mohammed V University in Rabat, Morocco., 4Thoracic Oncology Research Group, Laboratory Medicine and Molecular Pathology, Central Pathology Laboratory, St. James's Hospital, Dublin, Ireland Thoracic Oncology Research Group, Laboratory Medicine and Molecular Pathology, Central Pathology Laboratory, St. James's Hospital, Dublin, Ireland,
Objectives: Monoglyceride lipase (MGLL), as a prominent metabolic hub, is known to be actively involved in the development of the lipogenic phenotype, which promotes de novo lipid biosynthesis, allowing cancer cells to maintain their growth advantage, continuous proliferation, and metastasis. In this study, we aim to investigate mRNA MGLL expression levels and its sub-cellular localization in non-small cell lung cancer (NSCLC) cell lines, as well as examine the effect of two chemotherapy/targeted therapy agents, cisplatin and crizotinib, on the expression levels of MGLL. Methods: pcDNA3.1(-)-MGLL and pEGFPN1-MGLL constructs were sub-cloned into E.coli DH5a and transfected into NSCLC cell lines for MGLL expression evaluation and sub-cellular localization, using polymerase chain reaction (PCR) and quantitative PCR (qPCR) and fluorescence microscopy, respectively. Parent and cisplatin/crizotinib-resistant cell lines were grown and maintained in adequate media for subsequent MGLL expression analysis. Results: PCR and qPCR results revealed that MGLL was successfully transfected into the H1299 cells and efficiently expressed. Fluorescence microscopy of the pEGFPN1-MGLL transfected cells revealed a cytosolic expression of MGLL. As per the analysis on the effect of cisplatin and crizotinib on MGLL expression, a notable downregulation of MGLL expression was noted in the resistant cell lines. Conclusion: These results provide groundwork for further research on molecules modulating MGLL expression, which may be deemed helpful to provide therapy options targeting MGLL in NSCLC treatment. Keywords: Lung cancer, MGLL expression, pcDNA3.1, cisplatin
Cite This Article
Boustany Y, Laraqui A, Oukabli M, Sekhsokh Y, Belkadi B, Gray S. Molecular Cloning, Expression, and Sub-Cellular Localization of MGLL in NSCLC Cell Lines. EJMO. 2022; 6(4): 330-338
Corresponding Author: Youssra Boustany