A study on the gene expression level in HaCaT keratinocyte cells to relate with halal and haram status when exposed to plant and animal fats using cDNA microarray

Abstract

Halal and haram are ingrained in the daily life of a Muslim; guided by Al-Quran and As-Sunnah. This concerns of halal and haram has also opened a vast market operated by not only Muslims but non-Muslims all over the world. The rapid growth of halal market demands the use of technologies to ensure the quality and safety of halal products. These technologies range from compact and mobile test kits to the high-end techniques such as Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC) and Polymerase Chain Reaction (PCR). In line with the halal market growth, more scientific research related to halal by using the above techniques has also been reported. However, microarray has received very little attention in halal research. Therefore, this study explores the use of cDNA microarray to investigate the effects of fat from haram sources on HaCaT keratinocyte human skin cells in comparison to halal fat sources at gene expression level. The haram fat sources used in this study were lard and non-halal slaughtered lamb fat while Halal fat sources were virgin coconut oil (VCO) and halal slaughtered lamb fat. The RNAs extracted from treated cells were used in cDNA microarray (Agilent 8x60K SurePrint G3 Human GE). The data analyzed by GeneSpring GX 13.0, detected 50,739 genes from the four treatments and after further filtration; 53 genes were obtained with p-value of <0.05 and fold change of ?2.0 (FC range between -2.457 to 6.813). The most regulated genes were NLRP5, FABP3, RPS21, PRKDC, ERCC4, ACTG1P4, and RACGAP1P. Selected genes (FABP3, PRKDC, GULP1, and XPOT) were then validated using real-time PCR. The gene expressions from real-time PCR were found to be consistent with microarray data. Finally, pathway analysis using Ingenuity Pathway Analysis (IPA® 2016) software gave some insights into the underlying molecular networks and pathways. Although the results were not entirely conclusive, some patterns were observed; the four fat emulsion treatments were involved in similar bio functionalities (cellular growth and proliferation, cell cycle and cellular movement) and associated diseases (developmental disorders involving the cell growth, connective tissue and hematological disease). In conclusion, the study showed that halal and haram fat sources caused differential gene expression in human cells. However, more work is warranted to further elucidate the pathways involved in order to understand the potential benefits and/or the perceived harmful effects of the fats.