Deacetylase Plus Bromodomain Inhibition Downregulates ERCC2 and Suppresses the Growth of Metastatic Colon Cancer Cells

There is growing evidence that DNA repair factors have clinical value for cancer treatment. Nucleotide excision repair (NER) proteins, including excision repair cross-complementation group 2 (ERCC2), play a critical role in maintaining genome integrity. Here, we examined ERCC2 expression following epigenetic combination drug treatment. Attention was drawn to ERCC2 for three reasons. First, from online databases, colorectal cancer (CRC) patients exhibited significantly reduced survival when ERCC2 was overexpressed in colon tumors. Second, ERCC2 was the most highly downregulated RNA transcript in human colon cancer cells and rat tumors after treatment with the histone deacetylase 3 (HDAC3) inhibitor sulforaphane (SFN) plus JQ1, which is an inhibitor of the bromodomain and extraterminal domain (BET) family. Third, as reported here, RNA-sequencing of polyposis in rat colon (Pirc) polyps following treatment of rats with JQ1 plus 6-methylsulfinylhexyl isothiocyanate (6-SFN) identified Ercc2 as the most highly downregulated gene. The current work also defined promising second-generation epigenetic drug combinations with enhanced synergy and efficacy, especially in metastasis-lineage colon cancer cells cultured as 3D spheroids and xenografts. This investigation adds to the growing interest in combination approaches that target epigenetic ‘readers’, ‘writers’, and ‘erasers’ that are deregulated in cancer and other pathologies, providing new avenues for precision oncology and cancer interception.

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Cytotoxicity and Apoptosis Effects of Curcumin Analogue (2E,6E)-2,6-Bis(2,3-Dimethoxybenzylidine) Cyclohexanone (DMCH) on Human Colon Cancer Cells HT29 and SW620 In Vitro

Molecules ◽

10.3390/molecules26051261 ◽

2021 ◽

Vol 26 (5) ◽

pp. 1261

Author(s):

Nurul Fattin Che Rahim ◽

Yazmin Hussin ◽

Muhammad Nazirul Mubin Aziz ◽

Nurul Elyani Mohamad ◽

Swee Keong Yeap ◽

...

Keyword(s):

Colorectal Cancer ◽

Colon Cancer ◽

Cell Death ◽

Cancer Cells ◽

Cell Lines ◽

Curcuma Longa ◽

Metastatic Colon Cancer ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Sw620 Cell

Colorectal cancer (CRC) is the third most common type of cancer worldwide and a leading cause of cancer death. According to the Malaysian National Cancer Registry Report 2012–2016, colorectal cancer was the second most common cancer in Malaysia after breast cancer. Recent treatments for colon cancer cases have caused side effects and recurrence in patients. One of the alternative ways to fight cancer is by using natural products. Curcumin is a compound of the rhizomes of Curcuma longa that possesses a broad range of pharmacological activities. Curcumin has been studied for decades but due to its low bioavailability, its usage as a therapeutic agent has been compromised. This has led to the development of a chemically synthesized curcuminoid analogue, (2E,6E)-2,6-bis(2,3-dimethoxybenzylidine) cyclohexanone (DMCH), to overcome the drawbacks. This study aims to examine the potential of DMCH for cytotoxicity, apoptosis induction, and activation of apoptosis-related proteins on the colon cancer cell lines HT29 and SW620. The cytotoxic activity of DMCH was evaluated using the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) cell viability assay on both of the cell lines, HT29 and SW620. To determine the mode of cell death, an acridine orange/propidium iodide (AO/PI) assay was conducted, followed by Annexin V/FITC, cell cycle analysis, and JC-1 assay using a flow cytometer. A proteome profiler angiogenesis assay was conducted to determine the protein expression. The inhibitory concentration (IC50) of DMCH in SW620 and HT29 was 7.50 ± 1.19 and 9.80 ± 0.55 µg/mL, respectively. The treated cells displayed morphological features characteristic of apoptosis. The flow cytometry analysis confirmed that DMCH induced apoptosis as shown by an increase in the sub-G0/G1 population and an increase in the early apoptosis and late apoptosis populations compared with untreated cells. A higher number of apoptotic cells were observed on treated SW620 cells as compared to HT29 cells. Human apoptosis proteome profiler analysis revealed upregulation of Bax and Bad proteins and downregulation of Livin proteins in both the HT29 and SW620 cell lines. Collectively, DMCH induced cell death via apoptosis, and the effect was more pronounced on SW620 metastatic colon cancer cells, suggesting its potential effects as an antimetastatic agent targeting colon cancer cells.

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TGF-β1/PTEN/PI3K signaling plays a critical role in the anti-proliferation effect of tetrandrine in human colon cancer cells

International Journal of Oncology ◽

10.3892/ijo.2017.3875 ◽

2017 ◽

Vol 50 (3) ◽

pp. 1011-1021 ◽

Cited By ~ 11

Author(s):

Qian-Zhao Chen ◽

Yang Li ◽

Ying Shao ◽

Yu-Hua Zeng ◽

Wen-Yan Ren ◽

...

Keyword(s):

Colon Cancer ◽

Cancer Cells ◽

Critical Role ◽

Human Colon ◽

Pi3k Signaling ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Human Colon Cancer Cells ◽

Tgf Β1

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Expression of Kinase-defective Mutants of c-Src in Human Metastatic Colon Cancer Cells Decreases Bcl-xLand Increases Oxaliplatin- and Fas-induced Apoptosis

Journal of Biological Chemistry ◽

10.1074/jbc.m408550200 ◽

2004 ◽

Vol 279 (44) ◽

pp. 46113-46121 ◽

Cited By ~ 42

Author(s):

Gareth J. Griffiths ◽

Mei Yee Koh ◽

Valerie G. Brunton ◽

Christopher Cawthorne ◽

Natalie A. Reeves ◽

...

Keyword(s):

Colon Cancer ◽

Vector Control ◽

Cancer Cells ◽

Kinase Activity ◽

Src Kinase ◽

Human Colon ◽

Metastatic Colon Cancer ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Induced Apoptosis

Tumor resistance to current drugs prevents curative treatment of human colon cancer. A pressing need for effective, tumor-specific chemotherapies exists. The non-receptor-tyrosine kinase c-Src is overexpressed in >70% of human colon cancers and represents a tractable drug target. KM12L4A human metastatic colon cancer cells were stably transfected with two distinct kinase-defective mutants ofc-src.Their response to oxaliplatin, to SN38, the active metabolite of irinotecan (drugs active in colon cancer), and to activation of the death receptor Fas was compared with vector control cells in terms of cell cycle arrest and apoptosis. Both kinase-defective forms of c-Src co-sensitized cells to apoptosis induced by oxaliplatin and Fas activation but not by SN38. Cells harboring kinase-defective forms of c-Src carrying function blocking point mutations in SH3 or SH2 domains were similarly sensitive to oxaliplatin, suggesting that reduction in kinase activity and not a Src SH2-SH3 scaffold function was responsible for the observed altered sensitivity. Oxaliplatin-induced apoptosis, potentiated by kinase-defective c-Src mutants, was dependent on activation of caspase 8 and associated with Bid cleavage. Each of the stable cell lines in which kinase-defective mutants of c-Src were expressed had reduced levels of Bcl-xL.However, inhibition of c-Src kinase activity by PP2 in vector control cells did not alter the oxaliplatin response over 72 h nor did it reduce Bcl-xLlevels. The data suggest that longer term suppression of Src kinase activity may be required to lower Bcl-xLlevels and sensitize colon cancer cells to oxaliplatin-induced apoptosis.

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A Novel Mouse Model of Metastatic Colon Cancer Established by Endoscopic Injection of Human Colon Cancer Cells

Gastroenterology ◽

10.1016/s0016-5085(11)62830-8 ◽

2011 ◽

Vol 140 (5) ◽

pp. S-682

Author(s):

Oliver Movadat ◽

Alexandra Wlodarski ◽

Markus F. Neurath ◽

Christoph Becker

Keyword(s):

Colon Cancer ◽

Mouse Model ◽

Cancer Cells ◽

Human Colon ◽

Metastatic Colon Cancer ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Endoscopic Injection ◽

Human Colon Cancer Cells

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TDG interacts with DNMT3A to inhibit the migration and invasion of human colon cancer cells

10.21203/rs.3.rs-779154/v1 ◽

2021 ◽

Author(s):

Jiyu Miao ◽

Changan Zhao ◽

Kaijie Tang ◽

Xiaofan Xiong ◽

Fei Wu ◽

...

Keyword(s):

Colon Cancer ◽

Cancer Cells ◽

Excision Repair ◽

Human Colon ◽

Migration And Invasion ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Human Colon Cancer Cells

Abstract Background Colorectal cancer (CRC) is one of the most common malignant tumors with high recurrence and mortality. Thymine DNA glycosylase (TDG) is one of the key molecules involved in base excision repair pathway. Recently, more and more attentions have been paid to the role of TDG on tumor development. However, the specific functions of TDG in CRC remain unclear. Methods The biological functions of TDG and DNA methyltransferase 3 alpha (DNMT3A) in CRC were evaluated using migration and invasion assay. Tumor metastasis assay was performed in nude mice to detect the role of TDG in vivo. The interaction of TDG with DNMT3A was determined by co-immunoprecipitation (Co-IP). Chromatin immunoprecipitation analysis (CHIP) was applied to predict the DNA binding site of DNMT3A. We also performed methylation-specific PCR (MSP) to detect the changes in TIMP2 methylation levels. Results We found that TDG could inhibit the migration and invasion of human colon cancer cells in vitro and in vivo. TDG promoted the ubiquitination and degradation of DNMT3A by binding with it. Interference with siDNMT3A also inhibited the migration and invasion of human colon cancer cells. Further ChIP, MSP, and rescue experiments data confirmed that TDG accelerated the degradation of DNMT3A, and then significantly regulated the transcription and expression of TIMP2, thereby affecting the migration and invasion of human colon cancer cells. Conclusion Our findings reveal that TDG inhibit the migration and invasion of human colon cancer cells through DNMT3A-TIMP2 axis which may be potential therapeutic strategies in the development and treatment of CRC.

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Extravirgin olive oil up-regulates CB1 tumor suppressor gene in human colon cancer cells and in rat colon via epigenetic mechanisms

The Journal of Nutritional Biochemistry ◽

10.1016/j.jnutbio.2014.10.013 ◽

2015 ◽

Vol 26 (3) ◽

pp. 250-258 ◽

Cited By ~ 61

Author(s):

Andrea Di Francesco ◽

Anastasia Falconi ◽

Clara Di Germanio ◽

Maria Vittoria Micioni Di Bonaventura ◽

Antonio Costa ◽

...

Keyword(s):

Colon Cancer ◽

Olive Oil ◽

Cancer Cells ◽

Human Colon ◽

Suppressor Gene ◽

Rat Colon ◽

Epigenetic Mechanisms ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Human Colon Cancer Cells

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Influence of oxygen availability on expression of glutaminolysis genes in human colon cancer cells

Postępy Higieny i Medycyny Doświadczalnej ◽

10.2478/ahem-2021-0032 ◽

2021 ◽

Vol 75 (1) ◽

pp. 923-932

Author(s):

Dagmara Otto-Ślusarczyk ◽

Wojciech Graboń ◽

Magdalena Mielczarek-Puta ◽

Alicja Chrzanowska ◽

Anna Barańczyk-Kuźma

Keyword(s):

Colon Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Oxygen Availability ◽

Metabolic Adaptation ◽

Metastatic Colon Cancer ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Trypan Blue Exclusion ◽

Thiazolyl Blue Tetrazolium Bromide

Abstract Introduction Glutaminolysis, beside glycolysis, is a key metabolic pathway of a cancer cell that provides energy and substrates for the synthesis of nucleic acids, proteins, and lipids. The pathway is mediated by both mitochondrial and cytosolic enzymes. Neither expression of glutaminolysis enzymes in colon cancer cells nor the influence of various oxygen concentrations on their expression has been studied so far. Objectives The aim of the study was to determine and compare the mRNA expression of enzymes involved in glutaminolysis at various oxygen levels in human primary (SW480) and metastatic (SW620) colon cancer cells cultured in 1% O2 (hypoxia), 10% O2 (tissue normoxia), 21% O2 (atmospheric normoxia). Methods Cell viability was determined by Trypan Blue exclusion (TB) and Thiazolyl Blue Tetrazolium Bromide (MTT). The expression of HIF1α, GLUT1, GLS1, AST1, AST2, ACL, PC and GC1, GC2 at mRNA levelwas determined by RT-qPCR. Results. Correlation between increasing oxygen concentration and cell count was not observed. In both cell lines the number of viable cells was the lowest at 10% oxygen. The enzyme profile and expression of proteins involved in glutaminolysis varied depending on oxygen pressure and type of cell lines. In summary, our findings suggest differences in metabolic adaptation to oxygen availability in vivo between primary and metastatic colon cancer cells.

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