FTO rs9939609 and PPARy rs1801282 Polymorphisms in Mexican Adolescents With Overweight and Obesity at High Risk for Developing Diabetes

Study Population Students without known disease, between 14 and 18 years from the "Lic. Adolfo Lopez Mateos" Preparatory school of the Autonomous University of the State of Mexico (UAEMex), were invited to participate.

Anthropometry The body mass index (BMI) was calculated as weight (kg) divided by height (m) squared with conventional stadiometer; dividing the results in normal, overweight and obesity according to the classification of the World Health Organization (WHO), specifically for adolescents by gender and age.

Laboratory 3 mL of venous blood (Vacutainer tubes) were taken in fasting. The concentration of glucose, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides were determined by enzymatic methods (Randox Laboratories Ltd, County Antrim, UK); whereas leptin and adiponectin concentrations were determined by the immunosorbent assay (ELISA) (Invitrogen MR Frederyck MD USA). A sample remained frozen at -70°C (900 series Thermo Scientific) until DNA extraction.

Genotyping DNA extraction was performed in the Magna Pure LC 2.0 Instrument (Roche, Germany) using the MagNA Pure LC DNA Isolation Kit 1 (Roche, Germany). After extraction, the DNA was quantified using a NanoPhotometer (Implen GmbH, Germany), reporting concentration (μg/ml) and purity (260/280 absorbance). Genotyping was performed by polymerase chain reaction (PCR) Life express thermal cycler (Bioer, China). The primers and conditions for each polymorphism are listed in Table 1.

The oligonucleotides were designed using the Primer Quest web tool (Integrated DNA Technologies, Inc., CA, USA) and synthesized at the Synthesis and DNA Sequencing Unit of the National Autonomous University of Mexico (UNAM) Institute of Biotechnology (Cuernavaca, Morelos, Mexico). A search was performed in BLAST to verify the correct hybridization. The final products were visualized in 2% agarose gels, stained with ethidium bromide, and visualized under an UV transilluminator (Gel Logic 212 Pro, Carestream, USA). To confirm the detection of polymorphisms sequencing was performed at the National Institute of Genomic Medicine (INMEGEN), Mexico City.