Preliminary Study On The Function Of PPARG Gene In Human Cutaneous Squamous Cell Carcinoma Colo16 Cells

Background Cutaneous Squamous Cell Carcinoma(CSCC)is a skin malignancy that originates from epidermal or accessory keratinocytes.The incidence of CSCC accounts for about 20%of the skin malignancy and continues to increase.Up to date,the pathogenesis of CSCC is not yet clear.Peroxisome proliferator activated receptor γ(PPARy),encoded by the peroxisome proliferator activated receptor gama(PPARG)gene,is a ligand-dependent nuclear transcription factor and belongs to the nuclear hormone receptor superfamily.In recent years,the relationship between PPARy and tumor has attracting more and more attention.Studies have found that expression of PPARG is down-regulated in many tumor tissues.PPARγ activation by ligands can inhibit tumor cell proliferation,promote apoptosis,and inhibit tumor angiogenesis.So it is presumed that PPARG acts as a tumor suppressors in malignancy.Our previous study found that the expression of PPARG in CSCC was significantly reduced compared with paracancerous tissues.Based on this,we speculate that the PPARG gene may have the function of inhibiting CSCC formation.To verify our hypothesis,we intend to use gene silencing and overexpression techniques to preliminarily explore the function of PPARG gene in CSCC Colo16 cell line.Objective To preliminarily study the function of PPARG gene in CSCC Colo16 cell line.Methods 1.Colo 16 cells were divided into 5 groups:transfection-overexpressed PPARG plasmid group(Plasmid-PPARG),transfected control plasmid group(Plasmid-NC),transfected PPARG siRNA group(siR2),transfected control siRNA group(siR-NC),blank cell group(Blank).2.Using gene silencing and overexpression techniques,target components were respectively transfected into the above-mentioned groups.Total RNA and protein were extracted 48 hours after transfection,and the expression of PPARG mRNA and protein in each group of cells was detected by qPCR and Western Blotting,respectively.3.CCK-8 reagent was used to detect the change of cell proliferation ability in each group at different time points after transfection;flow cytometry was used to detect the changes of cell apoptosis in each group at 48 hours after transfection.Results Compared with Blank group,the expression of PPARG mRNA and protein in the Plasmid-PPARG group was significantly up-regulated(p<0.05),cell proliferation was down-regulated(p<0.05),cell apoptosis was increased with a significant difference(p<0.05).Compared with Blank cells,the expression of PPARG mRNA and protein in siR2 cells was significantly decreased(p<0.05),cell proliferation was enhanced(p<0.05),apoptosis was decreased(p<0.05).Conclusion PPARG gene can inhibit cell proliferation and promote apoptosis.PPARG may play the role of tumor suppressor gene in CSCC,but the function and mechanism as a tumor suppressor gene need further study.