*3.6. Functional Enrichment and Gene Ontology (GO) Analysis*

To better understand the molecular role of selected genes involved in the suppressive effect of Gemini-Cur on HCT-116 cells, DEGs were mapped in the GO database using an online web tool (https://maayanlab.cloud/Enrichr, 25 January 2021), and a threshold of Padj < 0.05 and gene counts > 5 was considered. Here, gene ontology was performed for 198 downregulated and 244 upregulated genes separately, and the status of all five selected genes in all three categories (BP; biological process, MF; molecular function, and CC; cellular component) was assessed.

Interestingly, the GO results of the integrated group showed that upregulated ATF3 is significantly enriched in BP category including response to endoplasmic reticulum stress, regulation of transcription from RNA polymerase II promoter in response to stress, and MF category with protein hetero-dimerization activity. In addition, GADD45G was enriched in the regulation of p38MAPK cascade as well as control of the p38MAPK pathway, both in BP process (Table 5). In contrast, downregulated genes (CCNA2, BUB1B, and CDK1) were involved in almost all three categories and different cellular cascades.
