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Microarray gene expression profiling in colorectal (HCT116) and hepatocellular (HepG2) carcinoma cell lines treated
Mohammad Faujul Kabir1, Johari Mohd Ali1, Onn Haji Hashim1
1Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Background:
We have previously reported anticancer activities of Melicope ptelefolia (MP) leaf extracts on four different cancer cell lines. However, the underlying mechanisms of actions have yet to be deciphered. In the present study, the anticancer activity of MP hexane extract (MP-HX) on colorectal (HCT116) and hepatocellular carcinoma (HepG2) cell lines was characterized through microarray gene expression profiling.
Methods:
HCT116 and HepG2 cells were treated with MP-HX for 24 hr. Total RNA was extracted from the cells and used for transcriptome profiling using Applied Biosystem GeneChip™ Human Gene 2.0 ST Array. Gene expression data was analysed using an Applied Biosystems Expression Console and Transcriptome Analysis Console software. Pathway enrichment analyses was performed using Ingenuity Pathway Analysis (IPA) software. The microarray data was validated by profiling the expression of 17 genes through quantitative reverse transcription PCR (RT-qPCR).
Results:
MP-HX induced differential expression of 1,290 and 1,325 genes in HCT116 and HepG2 cells, respectively (microarray data fold change, MA_FC ≥ ±2.0). The direction of gene expression change for the 17 genes assayed through RT-qPCR agree with the microarray data. In both cell lines, MP-HX modulated the expression of many genes in directions that support antiproliferative activity. IPA software analyses revealed MP-HX modulated canonical pathways, networks and biological processes that are associated with cell cycle, DNA replication, cellular growth and cell proliferation. In both cell lines, upregulation of genes which promote apoptosis, cell cycle arrest and growth inhibition were observed, while genes that are typically overexpressed in diverse human cancers or those that promoted cell cycle progression, DNA replication and cellular proliferation were downregulated. Some of the genes upregulated by MP-HX include pro-apoptotic genes (DDIT3, BBC3, JUN), cell cycle arresting (CDKN1A, CDKN2B), growth arrest/repair (TP53, GADD45A) and metastasis suppression (NDRG1). MP-HX downregulated the expression of genes that could promote anti-apoptotic effect, cell cycle progression, tumor development and progression, which include BIRC5, CCNA2, CCNB1, CCNB2, CCNE2, CDK1/2/6, GINS2, HELLS, MCM2/10 PLK1, RRM2 and SKP2. It is interesting to note that all six top-ranked genes proposed to be cancer-associated (PLK1, MCM2, MCM3, MCM7, MCM10 and SKP2) were downregulated by MP-HX in both cell lines.
Discussion:
The present study showed that the anticancer activities of MP-HX are exerted through its actions on genes regulating apoptosis, cell proliferation, DNA replication and cell cycle progression. These findings further project the potential use of MP as a nutraceutical agent for cancer therapeutics.
Insights
Melicope ptelefolia hexane extract (MP-HX) demonstrates anticancer effects by modulating genes involved in apoptosis, cell cycle, and proliferation. This study elucidates the molecular mechanisms behind MP-HX
Area of Science:
- Molecular Biology
- Pharmacology
- Genomics
Background:
- Previous studies reported anticancer activities of Melicope ptelefolia (MP) leaf extracts.
- The specific molecular mechanisms underlying MP's anticancer effects remained largely uncharacterized.
Purpose of the Study:
- To investigate the anticancer mechanisms of MP hexane extract (MP-HX) on colorectal (HCT116) and hepatocellular carcinoma (HepG2) cell lines.
- To characterize the effects of MP-HX on gene expression profiles using microarray analysis.
Main Methods:
- HCT116 and HepG2 cells were treated with MP-HX.
- Transcriptome profiling was performed using microarray gene expression analysis.
- Gene expression data was analyzed using specialized bioinformatics software, with validation via quantitative reverse transcription PCR (RT-qPCR).
Main Results:
- MP-HX significantly altered the expression of over 1,200 genes in both HCT116 and HepG2 cells.
- MP-HX modulated key pathways related to cell cycle, DNA replication, and cell proliferation, supporting antiproliferative activity.
- Upregulation of pro-apoptotic genes and downregulation of genes promoting cell cycle progression and cancer development were observed.
Conclusions:
- MP-HX exerts anticancer effects by regulating genes involved in apoptosis, cell proliferation, DNA replication, and cell cycle progression.
- These findings support the potential of MP as a nutraceutical agent for cancer therapy.
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