Table 3.
Isothiocyanate as epi-drugs against cancers.
| Bioactive Molecules |
Origin | Experimental Methods | Key Results | References |
|---|---|---|---|---|
| Moringa isothiocyanate | Not reported | Mouse epidermal JB6 P+ cell line Cell viability test: MTS assay RNA-seq DNA SureSelect Methyl-seq |
Induced the apoptosis of JB6 P+ cells in a dose- and time-dependent manner. Altered the gene expression profiles in mouse epidermal cells. Affected the canonical signaling pathways. Played a protective role during TPA-induced neoplastic/tumorigenic transformation in JB6 cells. Changed the DNA methylation during TPA-induced neoplastic/tumorigenic transformation in JB6 cells. Reversed the methylation changes in genes (hyper- or hypomethylation) that occur in a response to TPA. |
[16] |
| Phenethyl isothiocyanate (PEITC) | Purchased | PCa LNCaP and PC3 cell lines RNA isolation, miRNA profiling, and qPCR Transfection of has-miR-194-5p mimic and inhibitor Western blot analysis Luciferase reporter activity assay RNA interference |
Altered the miRNA expression in PCa cells miR-194 and suppressed PC3 cell invasiveness in vitro. Down-regulated the MMP2 and MMP9 via microRNA-194. Decreased BMP1 expression which decreased cellular MMP levels. |
[126] |
| Not reported | Colorectal cancer cell lines SW620, SW480, and HCT116 Cell viability analysis Western blot analysis RNA extraction and quantitative real-time PCR Epigenome-wide DNA methylation assays Cell cycle analysis |
Induced stable alterations in the expression profile of epigenetic writers/erasers and chromatin-binding of HDACs and Polycomb-group (PcG) proteins. PEITC exposure not only blocked HDAC binding to euchromatin but was also associated with hypomethylation of PcG target genes that are typically hypermethylated in cancer. Induced the expression of pro-apoptotic genes in tumor cells. |
[127] | |
| Purchased | PCa LNCaP cell line Setd7 knockdown in LNCaP cells RNA isolation Oligonucleotide microarray analyses for transcriptome profiling qPCR |
Impacted a large set of genes and caused a high fold change. Altered several signaling pathways, in particular inflammation-related TNFR signaling and PTEN/PI3K/AKT signaling. |
[128] | |
| Purchased | Human colon carcinoma cell line HT29 Cell viability assay Cell cycle arrest analysis Sprague Dawley rats Determination of DNMT1 and HDAC1 levels |
PEITC + laccaic acid (LA) reduced cell viability with apoptotic cell death. Induced necrotic cell death. PEITC + LA attenuated the number of aberrant crypt foci, DNMT1, and HDAC1 levels (in vivo). |
[129] | |
| Sulforaphane (SFN) | Not reported | Breast cancer MCF-7 and MDA-MB-231 cells Total RNA extraction, RT-PCR, and real-time quantitative PCR Western blot analysis HDAC, HAT, and DNMT activity assays Bisulfite sequencing analysis ChIP analysis Apoptosis assay |
Inhibited the viability and proliferation of breast cancer cells in vitro. Inhibited the hTERT in both breast cancer cells in a dose- and time-dependent manner. Decreased DNMT1 and DNMT3a in breast cancer cells. Increased the level of active chromatin markers acetyl-H3, acetyl-H3K9, and acetyl-H4. Decreased the trimethyl-H3K9 and trimethyl-H3K27 inactive chromatin markers in a dose-dependent manner. |
[130] |
| Purchased | Human colon adenocarcinoma cell lines Caco-2 and HCT116 Cell proliferation assay Genomic DNA and RNA isolation Quantification of gene-specific CpG island and global (LINE-1) methylation Real-time PCR |
No effect on the methylation of CpG islands in ESR1, p16INK4A or of LINE-1, a marker of global genomic methylation. Induced transient changes in DNMT mRNA expression. |
[131] | |
| Purchased | Ovarian cancer cell lines (SKOV3-ip1 and SKOV3TR-ip2 cells) MTT assay Clonogenic assay Analysis of apoptosis Western blot analysis Telomerase activity assay |
Inhibited cell viability of both ovarian cancer cells time- and dose-dependently. Arrested the ovarian cancer cells in the G2/M phase. Down-regulated Bcl-2 (a gene involved in anti-apoptosis) protein levels in both cell types. Up-regulated the cleaved poly(ADP-ribose) polymerase (PARP) and phosphorylated H2AX. SFN + EGCG arrested cells in both the G2/M and S phase. SFN + EGCG increased apoptosis in SKOV3TR-ip2 cells, while reducing the expression of hTERT. |
[132] | |
| Purchased | Prostate cancer TRAMP C1 cells DNA extraction Bisulfite genomic sequencing Methylation DNA immunoprecipitation (MeDIP) analysis RNA isolation and qPCR Western blot analysis ChIP assay |
Decreased methylated CpG ratio in the promoter region of Nrf2 gene in TRAMP C1 cells. Decreased the binding of anti-methyl cytosine antibody to the promoter region of Nrf2 gene in TRAMP C1 cells. Induced the expression of Nrf2 and its downstream gene. Altered the expressions of epigenetics modifying enzymes. |
[126] | |
| Purchased | Primary pancreatic ductal adenocarcinoma (PDA) cell line and non-malignant pancreatic ductal cells MTT assay Apoptosis measurement Detection of ALDH1 activity Western blot analysis Detection of MMP-2, MMP-9, and K-ras mRNA expression Detection of microRNA expression |
Inhibited colony formation. Sulforaphane + EGCG inhibited viability, migration, expression of MMP-2 and -9, ALDH1 activity, colony, and spheroid formation and induced the apoptosis. Sulforaphane + EGCG induced the expression of miR-let7-a in cancer cells. |
[133] | |
| Purchased | JB6 P+ cells Western blot analysis RNA isolation Quantitative real-time PCR DNA isolation Bisulfite genomic sequencing HDAC activity assay |
Increased Nrf2 nuclear translocation and protein expression, up-regulating the mRNA and protein expression of Nrf2 target enzymes in JB6 P+ cells. Inhibited TPA-induced JB6 P+ cell transformation. Increased relative Nrf2 mRNA expression when cells are treated with TPA. Decreased the methylation status of the Nrf2 gene promoter. Down-regulated the epigenetic modifying enzymes in JB6 P+ cells. |
[134] | |
| Purchased | Prostate cancer cells (LnCAP and PC3) ChIP assay Sample preparation for DNA methylation array DNA methylation array data analysis Validation of DNA methylation data Gene expression analyses |
SFN + DIM decreased DNMT gene expression and caused distinct DNA methylation profile alterations depending on prostate cell line. SFN + DIM reversed cancer-associated DNA methylation alterations in LnCAP cells. |
[16] | |
| Purchased | Cervical carcinoma cell line (HeLa) DNMT and HDAC activity assay In Silico molecular modeling studies of DNMT3B and HDAC1 Bisulfite Modification and MSP RT-PCR |
Inhibited the DNMTs activity and down-regulated the expression of DNMT3B in HeLa cells. Inhibited the HDACs activity and reduced the expression of HDAC1 in HeLa cells. Interacted with DNMT3B and HDAC1. Reactivated or increased the expression of RARβ, CDH1, DAPK1, and GSTP1 genes in HeLa cells. |
[135] | |
| Purchased | Breast cancer cells (MCF-7 and MDA-MB-231) Cell growth and apoptosis assays Methylation-sensitive restriction analysis Quantitative real-time PCR |
Inhibited breast cancer cell growth and enhanced the clofarabine (ClF) inhibitory effect on cell viability. Induced the apoptosis in breast cancer cells and enhanced ClF Pro-apoptotic effects. Induced the hypomethylation of PTEN and RARbeta2 promoters with concomitant gene up-regulation. Increased epigenetic effects of ClF at non-invasive stages of breast cancer development. Induced p21 without altering DNMT1 expression. |
[91] | |
| Purchased | Human lung cancer cells (A549 and H1299) HDAC activity assay Western blot analysis qRT-PCR Apoptosis assay Cell cycle analysis Tumor xenografts |
Inhibited HDAC activity in lung cancer cells. Increased the acetylation of histones H3 and H4. Increased cell cycle and apoptotic-related gene expressions. Induced cell death and inhibited cell cycle arrest at S and G2/M in lung cancer cells. Suppressed the growth of A549 xenografts and HDAC activity in vivo. |
[136] | |
| Purchased | Human breast cancer cells MCF-7, MDA-MB-231, and SK-BR-3 MTT assay Cell cycle analysis Apoptosis assay Genotoxicity and DNA damage response Western blot analysis Real-time PCR Global DNA methylation N6-methyladenosine (m6A) RNA methylation microRNA profiling |
Promoted cell cycle arrest, elevation in the levels of p21 and p27, and cellular senescence (5–10 μM). Induced apoptosis (at 20 μM) Stimulated the energy stress as judged by decreased pools of ATP and AMPK activation and autophagy induction. Induced the global DNA hypomethylation. Decreased the levels of DNMT1 and DNMT3B. Diminished pools of m6A RNA methylation. Decreased the levels of miR-23b, miR-92b, miR-381, and miR-382 in breast cancer cells. |
[137] | |
| Not reported | Breast cancer cells (MCF-7 and MDA-MB-231) Cell density assay MTT assay RNA isolation qRT-PCR Western blot analysis DNMTs and HDACs activity assay |
SFN + WA promoted cell death. SFN + WA decreased the HDAC expression and promoted varying changes in DNMT expression. SFN + WA induced the changes in BAX and BCL-2. |
[138] | |
| Purchased | Breast cancer cell line MCF-7 ChIP assay UPLC-Orbitrap-MS Immunofluorescence assay |
Restored the alanine and lactic acid levels. Increased the levels of 4-OCH3E2. Influenced the expression of COMT through methylation mechanisms. Reversed the E2-induced methylation status. Epigenetic modulation of COMT expression subsequently influenced E2 metabolism. |
[139] | |
| Purchased | Lung adenocarcinoma (A549), embryonic kidney (HEK293), and colorectal adenocarcinoma (HT29) cell lines Cell viability assay Bisulfite genome sequencing MeDIP analysis RNA extraction Quantitative real-time PCR Western blot analysis ChIP assay HDAC and DNMT activity assays |
Decreased CpG methylation in the promoter region of miR-9-3. Increased H3K4me1 enrichment at the miR-9-3 promoter. Induced miR-9-3 expression. Diminished the expression and activity of epigenetic modifying enzymes. Decreased the protein expression of CDH1. |
[140] | |
| Not reported | Human breast cancer cells Quantitative real-time PCR Western blot analysis HDACs activity assay Global histone H3 acetylation quantification Global DNA methylation analysis RNA sequencing analysis |
SFN-based broccoli sprout diet induced: Decreased tumor incidence and inhibited early breast cancer development. Increased gene transcription in tumor suppressor genes such as p53 and p16INK4a. Decreased expressions of tumor-promoting genes such as TERT and c-Myc. Decreased gene expression and enzymatic activity of HDAC1, but did not affect DNMT1 gene expression. Decreased global DNA methylation level. Increased histone acetylation levels in two important histone acetylation markers (histone acetyl-H3K9 and acetyl-H3K14). |
[141] | |
| Not reported | Human breast cancer MCF-7 and MDA-MB-231 cells Cell viability assay Methylation-sensitive restriction analysis Quantitative real-time PCR (MSRA) |
SFN + clofarabine reactivated the DNA methylation- silenced CDKN2A tumor suppressor. SFN + clofarabine inhibited cancer cell growth at a non-invasive breast cancer stage. |
[142] | |
| Not reported | Colon cancer (HCT116 and RKO) cells MTT assay Cellular density assay RT-PCR Cell cycle analysis Western blot assay HDAC activity assay |
Decreased cell density. Inhibited the cell viability. Induced the apoptosis. Down-regulated the oncogenic miR-21, HDAC and hTERT mRNA and protein and enzymatic levels in colorectal cancer cells. |
[143] | |
| Purchased | Breast cancer cells (MCF-7 and MDA-MB-231) MTT assay RNA isolation Quantitative real-time PCR Cell cycle analysis Western blot analysis HDAC activity assay Histone methyltransferase (HMT) activity assay |
SFN + GEN decreased cell viability of breast cancer cell lines. SFN + GEN increased the rate of apoptosis and lowered the colony formation potential of cells. SFN + GEN inhibited cell cycle progression to the G2 phase in MDA-MB-231 and G1 phase in MCF-7 breast cancer cell lines. SFN + GEN inhibited HDAC and HMT. SFN + GEN down-regulated the levels of HDAC2 and HDAC3 both at the mRNA and protein levels. SFN + GEN down-regulated the hTERT levels. |
[144] | |
| Purchased | Breast cancer cell lines (MCF-7 and MDA-MB-231) Flow cytometry cell cycle analysis DNA extraction Quantitative RT-PCR Western blot analysis Global methylation activity assay Histone acetyltransferase activity/Inhibition assay Histone methyltransferase activity/Inhibition assay ChIP assay |
SFN + WA regulated cell cycle progression from the S to the G2 phase through the inhibition of cell cycle genes in breast cancer cells. SFN + WA promoted changes in epigenetic regulators in MCF-7 and MDA-MB-231 cells. SFN + WA promoted changes in p53 and p21 in breast cancer cells. |
[138] | |
| Not reported | Nasopharyngeal carcinoma (NPC) cells, C666-1 Cancer stem cell (CSC) Cell viability analysis Flow cytometric analysis Real-time PCR Small interfering RNA (siRNA)-mediated silencing method Western blot analysis Nude mice tumorigenicity assay |
Inhibited the formation of CSC-enriched NPC tumor spheres. Reduced the population of cells with CSC-associated properties. Restored the expression of Wnt inhibitory factor 1 (WIF1). Down-regulated DNMT1 activity. Inhibited the in vivo growth of C666-1 cells. Enhanced the anti-tumor effects of cisplatin. |
[145] | |
| Not reported | Mouse melanoma B16F10 cells Cell cycle arrest analysis Apoptotic analysis RNA-Seq analysis Histone PTM mass spectrometry |
SFN and DAC single and combination treatment resulted in growth inhibition. SFN and DAC single and combination treatment resulted in minimal apoptosis. SFN and DAC single and combination treatment resulted in no cell cycle arrest. SFN induced dysregulated gene transcription. SFN and DAC uniquely induced dysregulated gene transcription. No significant alterations detected in histone epigenetic modifications. |
[146] | |
| Not reported | Neural crest cells (NCCs), JoMa1.3 cells Quantitative real-time PCR Western blot analysis ChIP-qRT-PCR analysis Snail1 siRNA transfection Analysis of apoptosis |
Reduced ethanol-induced apoptosis. Diminished ethanol-induced changes in the expression of E-cadherin and vimentin. Restored the EMT (epithelial-mesenchymal transition) in ethanol-exposed NCCs. Diminished the ethanol-induced reduction of H3K4me3 at the promoter regions of the Snail1 gene. Restored the expression of Snail1. Down-regulated the Snail1 target gene E-cadherin. |
[147] | |
| Not reported | Hepatocarcinoma cells, HepG2 and human primary gastric cells, GAS RNA-Seq analysis Cell cycle arrest analysis |
Induced cytotoxic effects and reduced the cell viability in both cell lines at higher concentrations. Induced cell cycle arrest in G2/M. Increased the expression of cyclin-dependent cyclins and kinases (CDK). Induced DNA damage in HepG2. Inhibited HDACs activity. Down-regulated the chromatin profile controlling enzymes. Induced the apoptosis and triggered pro-apoptotic signals in both cell lines. Down-regulated the MAPK/ERK/JUN and PIK3/AKT signaling pathways. Altered the methylation pattern in both strains. |
[148] | |
| Purchased | Caco-2 human colon adenocarcinoma cells DNA extraction and C-T conversion MSP RT-qPCR DNMT1 enzyme activity detection Western blot analysis |
No effect on the DNMT1 mRNA expression levels. SFN and 5-Aza + TSA inhibited DNMT1 protein expression. Decreased Nrf2 promoter methylation. Inhibited Nrf2 protein expression. |
[149] | |
| Purchased | Human hepatocarcinoma cell line, HepG2 Cell viability assays RNA extraction RNA-Seq analysis Apoptosis detection by flow cytometry Cell cycle analysis by flow cytometry |
Reduced the viability of HepG2 cells. Induced DNA damage, mitotic spindle abnormalities, apoptosis, and proliferation inhibition in HepG2 cells. Up-regulated the DNA damage response and cell cycle checkpoint genes. Down-regulated the pathways frequently overexpressed in human cancer. Inhibited HDACs activity. Affected the activity of oncogenic TF through methylation of its binding sites motifs. |
[150] |