Effect of Garlic Organic Sulfides on Gene Expression Profiling in HepG2 Cells and Its Biological Function Analysis by Ingenuity Pathway Analysis System and Bio-Plex-Based Assays

Abstract

Garlic organic sulfides are dietary bioactive components with multiple biofunctions to prevent chronic diseases/inflammation and promote human health. DADS (diallyl disulfide), DATS (diallyl trisulfide), and DTS (diallyl tetrasulfide) are typical organic sulfides with similar structures from garlic. However, the structure-activity relationship of garlic organic sulfides remained unknown. The aim of the present study was to investigate the effect of DADS, DATS, and DTS on the gene expression profiling of human hepatocellular carcinoma cells (HepG2) by application of microarray and specialized analysis software, GO, Bio-Plex-based cytokines assay and IPA and analyze their structure-activity relationship according to antioxidant, anti-inflammatory, and metabolic-related properties. According to the microarray data, with the increase of S atom in garlic organic sulfides, its biological activity was gradually enhanced. In the general catalog of GO, garlic organic sulfides mainly affect biological process, molecular function, and cellular component. RT-qPCR results indicated that the microarray data is trustworthy, and the structure-activity analysis data found that more sulfur atoms have more powerful properties; thus, microarray data of DTS was preceded to the subsequent IPA analysis. The results of IPA analysis showed that the top 5 signaling pathways and molecular functions were disturbed by DTS; the molecular functions with the highest scores affected by DTS are cancer, cell apoptosis, and cell proliferation, which imply that the occurrence or metabolism of these diseases is related to the differential expression of the above-mentioned related genes and the activation of signaling channels, and the core of the most significant molecular network is inflammation. Finally, the results found that the secretions of 6 cytokines in macrophages were significantly inhibited by DTS treatment. This is the first study that analyzed the structure-activity relationship of garlic organic sulfides, which will provide useful genetic information for its multi-biofunction and promote their clinical application in the near future.

Role of Sulfur Compounds in Vegetable and Mushroom Aroma

“…Sulfides are also associated with the off-flavors of many other products, such as drinking water, alcoholic beverages, and juices [80][81][82][83]. The sensory properties of sulfides are similar to each other because these compounds are most often characterized by a pungent smell and a spicy taste [26,84,85].…”

Garlic‐derived compounds: Epigenetic modulators and their antitumor effects

“…Other sulfides, such as SAC, SAMC, Ajoene, and S-methyl-cysteine sulfoxide, combine with DADS and DATS to inhibit tumor cell proliferation by inducing human cell apoptosis (Zhou et al, 2022). The antitumor effect of organic sulfide in garlic is not limited to specific tissues or carcinogens, mainly dependent on allyl and sulfhydryl, and enhanced with the increase of the number of sulfur atoms (C. Lv et al, 2021).…”

“…Relevant research reports show that the antitumor effect of DATS is better than DADS. According to microarray data, with the increase of S atoms in garlic organic sulfide, its biological activity gradually increased (C. Lv et al, 2021). The effects of ajoene extract, a garlic‐derived compound, on the levels of IL‐6 and myostatin in the muscles of tumor‐bearing mice.…”

“…Previous studies suggested that garlic‐derived compounds were more involved in epigenetic regulatory pathways as histone deacetylase inhibitors. Diallyl trisulfide (DATS) or diallyl disulfide (DADS) combined with docetaxel delivered a synthetic antitumor effect, in which natural products garlic enhances the chemosensitivity of cancer cells to docetaxel through a histone acetylation pathway (C. Lv et al, 2021; Pan et al, 2016). However, recent results have shown that garlic‐derived compounds DATS in cooperation with DNA methyltransferase (DNMT) inhibitors elicit synergistic efficiency in the demethylation of some “tough” CpG sites.…”

“…These “tough” CpG sites are usually in important regulatory regions (e.g., DMRs) for tissue‐specific gene expression, such as the transcription factor target motifs within CpGI (CpG island), where hypermethylation impairs gene transcription and is difficult to be erased by DNMT inhibitors alone (Ehrlich, 2019; Sae‐Lee et al, 2022). Garlic‐derived compounds may be more specifically effective in cancer cells with epigenetic dysregulation, differing from the current chemotherapeutic agents with both strong antitumor and toxic effects (C. Lv et al, 2021). The combination of garlic‐derived compounds with epigenetic inhibitors or chemotherapeutic drugs thus may be a more efficient （related to 5‐Aza‐CdR）(Lin et al, 2022; Lin et al, 2023) and safer (compared to first‐line chemotherapy agents） (Pan et al, 2016) therapeutic option in some cancer subtypes (i.e., personalized treatment).…”