The Effects of 5‑Aza‑2′‑Deoxycytidine and Valproic Acid on Apoptosis Induction and Cell Growth Inhibition in Colon Cancer HT 29 Cell Line

Masumeh Sanaei, Fraidoon Kavoosi, Hamed Sahraeian

Abstract


Background: Epigenetic changes, including DNA methylation and histone modification, alter gene expression without the nucleotide template alterations and are associated with all stages of tumor formation and progression. Previously, we investigated the effects of DNA demethylating agents and histone deacetylase inhibitors on hepatocellular carcinoma and colon cancers. The current study aimed to investigate the effects of 5‑aza‑2′‑deoxycytidine (5‑AZA‑CdR, decitabine) and valproic acid (VPA), individually as well as combined on apoptosis induction and cell growth inhibition in colon cancer HT 29 cell line.

Methods: The effect of the compounds on the cell viability was measured by MTT assay. To determine cell apoptosis, the cells were treated with 5‑aza‑CdR and VPA. Propidium iodide was used for staining and the cells were analyzed using flow cytometry.

Results: Both agents decreased cell viability in a time and dose‑dependent manner significantly (P < 0.002). The results of flow cytometry demonstrated that 5‑aza‑CdR and VPA induced apoptosis significantly as opposed to control groups. Maximal percentage of apoptotic cells was obtained after 48 h with combined treatment.

Coclusions: Our findings suggest that 5‑aza‑CdR and VPA can significantly inhibit cell growth and induce apoptosis in colon cancer HT 29 cell line.


Keywords


Apoptosis; colonic neoplasms; decitabine; valproic acid

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References


Ng J, Yu J. Promoter hypermethylation of tumour suppressor

genes as potential biomarkers in colorectal cancer. Int J Mol Sci

;16:2472‑96.

Grønbæk K, Treppendahl M, Asmar F, Guldberg P. Epigenetic

changes in cancer as potential targets for prophylaxis

and maintenance therapy. Basic Clin Pharmacol Toxicol

;103:389‑96.

Heichman KA, Warren JD. DNA methylation biomarkers and

their utility for solid cancer diagnostics. Clin Chem Lab Med

;50:1707‑21.

Delpu Y, Cordelier P, Cho W, Torrisani J. DNA methylation and

cancer diagnosis. Int J Mol Sci 2013;14:15029‑58.

Ropero S, Esteller M. The role of histone deacetylases (HDACs)

in human cancer. Mol Oncol 2007;1:19‑25.

Gnyszka A, Jastrzebski Z, Flis S. DNA methyltransferase

inhibitors and their emerging role in epigenetic therapy of cancer.

Anticancer Res 2013;33:2989‑96.

Singh V, Sharma P, Capalash N. DNA methyltransferase‑1

inhibitors as epigenetic therapy for cancer. Curr Cancer Drug

Targets 2013;13:379‑99.

Robertson KD, A. Jones P. DNA methylation: Past, present and

future directions. Carcinogenesis 2000;21:461‑7.

Momparler RL. Epigenetic therapy of cancer with

‑aza‑2′‑deoxycytidine (decitabine). Seminars in Oncology. 32.

Elsevier; 2005. p. 443‑51.

Federico M, Bagella L. Histone deacetylase inhibitors in the

treatment of hematological malignancies and solid tumors.

Biomed Res Int 2010;2011:1-12.

Xu W, Parmigiani R, Marks P. Histone deacetylase inhibitors:

Molecular mechanisms of action. Oncogene 2007;26:5541.

Chavez‑Blanco A, Perez‑Plasencia C, Perez‑Cardenas E,

Carrasco‑Legleu C, Rangel‑Lopez E, Segura‑Pacheco B,

et al. Antineoplastic effects of the DNA methylation inhibitor

hydralazine and the histone deacetylase inhibitor valproic acid in

cancer cell lines. Cancer Cell Int 2006;6:2.

Kavoosi F, Sanaei M. Comparative analysis of the effects of

valproic acid and tamoxifen on proliferation, and apoptosis of

human hepatocellular carcinoma WCH 17 CellLin. Iran J Ped

Hematol Oncol 2018;8:12‑20.

Sanaei M, Kavoosi F, Mansoori O. Effect of valproic acid

in comparison with vorinostat on cell growth inhibition and

apoptosis induction in the human colon cancer SW48 cells

in vitro. Exp Oncol 2018;40:95‑100.

Sanaei M, Kavoosi F, Roustazadeh A, Shahsavani H.

In vitro effect of the histone deacetylase inhibitor valproic

acid on viability and apoptosis of the PLC/PRF5 human

hepatocellular carcinoma cell line. Asian Pac J Cancer Prev

;19:2507‑10.

Sanaei M, Kavoosi F, Salehi H. Genistein and trichostatin A

induction of estrogen receptor alpha gene expression, apoptosis

and cell growth inhibition in hepatocellular carcinoma HepG

cells. Asian Pac J Cancer Prev 2017;18:3445.

Sanaei M, Kavoosi F, Atashpour S, Haghighat S. Effects of

genistein and synergistic action in combination with tamoxifen

on the HepG2 human hepatocellular carcinoma cell line. Asian

Pac J Cancer Prev 2017;18:2381.

Sanaei M, Kavoosi F, Pourahmadi M, Moosavi SN. Effect of

genistein and 17‑β estradiol on the viability and apoptosis of

human hepatocellular carcinoma HepG2 cell line. Adv Biomed

Res 2017;6:163.

Sanaei M, Kavoosi F, Roustazadeh A, Golestan F. Effect of

Genistein in comparison with trichostatin A on reactivation of

DNMTs genes in hepatocellular carcinoma. J Clin Transl Hepatol

;6:1‑6.

Sanaei M, Kavoosi F, Valiani A, Ghobadifar MA. Effect of

genistein on apoptosis and proliferation of hepatocellular

Carcinoma Hepa1‑6 Cell Line. Int J Prev Med 2018;9:12.

Okugawa Y, Grady WM, Goel A. Epigenetic alterations in

colorectal cancer: Emerging biomarkers. Gastroenterology

;149:1204‑25. e12.

Agrawal K, Das V, Vyas P, Hajdúch M. Nucleosidic DNA

demethylating epigenetic drugs–A comprehensive review from

discovery to clinic. Pharmacol Ther 2018;188:45‑79.

Chen HP, Zhao YT, Zhao TC. Histone deacetylases and

mechanisms of regulation of gene expression. Crit Rev Oncog

;20:35‑47.

Schneider‑Stock R, Diab‑Assef M, Rohrbeck A,

Foltzer‑Jourdainne C, Boltze C, Hartig R, et al. RETRACTION:

‑aza‑cytidine is a potent inhibitor of DNA methyltransferase

a and induces apoptosis in HCT‑116 colon cancer cells via

Gadd45‑and p53‑dependent mechanisms. J Pharmacol Exp Ther

;312:525‑36.

Li X, Qin B, Liu B. Delineating the effect of demethylating agent

‑aza‑2′‑deoxycytidine on human Caco‑2 colonic carcinoma

cells. Oncol Lett 2016;12:139‑43.

Rawłuszko‑Wieczorek AA, Horbacka K, Krokowicz P,

Misztal M, Jagodziński PP. Prognostic potential of DNA

methylation and transcript levels of HIF1A and EPAS1 in

colorectal cancer. Mol Cancer Res 2014;12:1112‑27.

Papi A, Ferreri A, Guerra F, Orlandi M. Anti‑invasive effects and

proapoptotic activity induction by the rexinoid IIF and valproic

acid in combination on colon cancer cell lines. Anticancer Res

;32:2855‑62.

Mologni L, Cleris L, Magistroni V, Piazza R, Boschelli F,

Formelli F, et al. Valproic acid enhances bosutinib cytotoxicity in

colon cancer cells. Int J Cancer 2009;124:1990‑6.

Yang H, Hoshino K, Sanchez‑Gonzalez B, Kantarjian H,

Garcia‑Manero G. Antileukemia activity of the combination of

‑aza‑CdR with valproic acid. Leukemia Res 2005;29:739‑48.

Gore SD, Baylin S, Sugar E, Carraway H, Miller CB,

Carducci M, et al. Combined DNA methyltransferase and histone

deacetylase inhibition in the treatment of myeloid neoplasms.

Cancer Res 2006;66:6361‑9.

Wischnewski F, Pantel K, Schwarzenbach H. Promoter

demethylation and histone acetylation mediate gene expression

of MAGE‑A1,‑A2,‑A3, and‑A12 in human cancer cells. Mol

Cancer Res 2006;4:339‑49.