Expression of diabetes-related genes in rodent β-cell line (RIN-m) treated with monocarbonyl curcumin analogues (C66 and B2BrBC) in presence or absence of streptozotocin – PCR array data

Published: 31 March 2022| Version 1 | DOI: 10.17632/vdgz7pk6vh.1
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Description

Curcumin monocarbonyl analogues C66* and B2BrBC** were synthesized at the Institute of Chemistry, Ss. Cyril and Methodius University in Skopje, Macedonia by substituting benzaldehyde with cyclohexanone in a base catalyzed aldol reaction. RIN-m rat β-cells (ATCC, Cat. # CRL-2057) were grown in RPMI-1640 medium (ATCC, Cat. # 30-2001) supplemented with 10% FBS (VWR, Cat. # 89510-186) and antibiotic/antimycotic mixture (MP Biomedicals, Cat. # 1674049) in incubator with standard conditions (5% CO2 and temperature of 37 °C). For experiments, 0.3 x 10(6) cells were seeded in 6-well plates with tissue culture growth media. The following day the media was replaced with serum-free media supplemented with 1.5 mM of streptozotocin (Sigma-Aldrich Cat. # 85882), 50 µM of C66, or B2BrBC dissolved in DMSO (VWR, Cat. # N182), and combination of streptozotocin with C66 or B2BrBC, for 72 hours. Total RNA was extracted using TRIzol reagent (Ambion, Cat. # 15596018). The quantification of RNA was made using NanoDrop One spectrophotometer (Thermo Fisher Scientific) after which the samples were normalized to 1 µg total RNA and converted to cDNA using qScript cDNA SuperMix (Quantabio, Cat. # 95048). PCR array was performed using QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) and RT2 Profiler PCR Array – Rat Diabetes (Qiagen, Cat. # 330231, GeneGlobe ID PARN-023ZA). The Ct values were obtained using QuantStudio Design & Analysis Software. * (2E,6E)-2,6-bis[(2-trifluoromethyl)benzylidene]cyclohexanone, C22H16F6O ** (2E,6E)-2,6-bis(2-bromobenzylidene)cyclohexanone), C20H16Br2O

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Institutions

Lenox Hill Hospital, Saints Cyril and Methodius University in Skopje Faculty of Natural Sciences and Mathematics, Feinstein Institute for Medical Research, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Northwell Health

Categories

Diabetes, Animal, Rat, Curcumin, Pancreas, Cell Culture, Animal Cell Culture, Polymerase Chain Reaction, Data Array, Rodent, Beta Cell, Analog, Real-Time Polymerase Chain Reaction, In Vitro Screening, In Vitro Study, Animal Diabetes, Reverse Transcription Polymerase Chain Reaction, Quantitative Reverse Transcription Polymerase Chain Reaction

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