Molecular Cloning, Expression, and Sub-Cellular Localization of MGLL in NSCLC Cell Line
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.
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