Genome-wide transcriptome analysis reveals inactivation of Wnt/β-catenin by 3,3'-diindolylmethane inhibiting proliferation of colon cancer cells

Leem,Sun-Hee; Li,Xiu   Juan; Park,Man  Hee; Park,Byung  Hyun; Kim,Soo  Mi

doi:10.3892/ijo.2015.3089

Genome-wide transcriptome analysis reveals inactivation of Wnt/β-catenin by 3,3'-diindolylmethane inhibiting proliferation of colon cancer cells

Authors:
- Sun-Hee Leem
- Xiu Juan Li
- Man Hee Park
- Byung Hyun Park
- Soo Mi Kim
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Affiliations: Department of Biological Science, Dong-A University, Busan 602-760, Republic of Korea, Department of Physiology, Institute for Medical Sciences, Chonbuk National University Medical School, Jeonju 561-180, Republic of Korea, Catholic University of Pusan, Busan 609-757, Republic of Korea, Department of Biochemistry, Institute for Medical Sciences, Chonbuk National University Medical School, Jeonju 561-180, Republic of Korea
Published online on: July 17, 2015 https://doi.org/10.3892/ijo.2015.3089
Pages: 918-926

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Abstract

Multiple genetic and signaling pathway alterations underlie the development of colon cancer. We utilized genome-wide transcriptome analysis to identify important gene expression patterns following treatment with 3,3'-diindolylmethane (DIM), a natural compound derived from cruciferous vegetables, on colon cancer cells. Statistical analyses of gene expression data from DIM treated cells revealed that 692 genes were significantly upregulated, while 731 genes were downregulated. Putative gene networks showed that several oncogenes (β-catenin, Myc and FOS) were significantly suppressed by DIM treatment. Using clinical data from colon cancer patients, activation of β-catenin was found to be significantly associated with patient prognosis by Kaplan-Meir plot analysis. We validated the mRNA and protein expression levels of c-Myc, β-catenin, and cyclin D1, all of which were significantly suppressed after DIM treatment in DLD-1 and HCT116 cells. System level characterization of our findings suggests for the first time that β-catenin and c-Myc, which are major genes involved in colon carcinogenesis, were significantly downregulated by DIM treatment in colon cancer cells. Therefore, targeting Wnt/β-catenin signaling by DIM may be an attractive strategy for the prevention and treatment of colon cancer.

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1	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Center MM, Ward E and Thun MJ: Cancer occurrence. Methods Mol Biol. 471:3–29. 2009. View Article : Google Scholar
3	DeSantis CE, Lin CC, Mariotto AB, Siegel RL, Stein KD, Kramer JL, Alteri R, Robbins AS and Jemal A: Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin. 64:252–271. 2014. View Article : Google Scholar : PubMed/NCBI
4	Li XJ, Leem SH, Park MH and Kim SM: Regulation of YAP through an Akt-dependent process by 3, 3′-diindolylmethane in human colon cancer cells. Int J Oncol. 43:1992–1998. 2013.PubMed/NCBI
5	Cho KH, Park S, Lee KS, Jang SI, Yoo KB, Kim JH and Park EC: A single measure of cancer burden in Korea from 1999 to 2010. Asian Pac J Cancer Prev. 14:5249–5255. 2013. View Article : Google Scholar : PubMed/NCBI
6	Park JH, Lee KS and Choi KS: Burden of cancer in Korea during 2000–2020. Cancer Epidemiol. 37:353–359. 2013. View Article : Google Scholar : PubMed/NCBI
7	Huguet EL, McMahon JA, McMahon AP, Bicknell R and Harris AL: Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54:2615–2621. 1994.PubMed/NCBI
8	Weber-Hall SJ, Phippard DJ, Niemeyer CC and Dale TC: Developmental and hormonal regulation of Wnt gene expression in the mouse mammary gland. Differentiation. 57:205–214. 1994. View Article : Google Scholar : PubMed/NCBI
9	Moon RT and Miller JR: The APC tumor suppressor protein in development and cancer. Trends Genet. 13:256–258. 1997. View Article : Google Scholar : PubMed/NCBI
10	Takahashi M, Fukuda K, Sugimura T and Wakabayashi K: Beta-catenin is frequently mutated and demonstrates altered cellular location in azoxymethane-induced rat colon tumors. Cancer Res. 58:42–46. 1998.PubMed/NCBI
11	Bienz M and Clevers H: Linking colorectal cancer to Wnt signaling. Cell. 103:311–320. 2000. View Article : Google Scholar : PubMed/NCBI
12	Luu HH, Zhang R, Haydon RC, Rayburn E, Kang Q, Si W, Park JK, Wang H, Peng Y, Jiang W, et al: Wnt/beta-catenin signaling pathway as a novel cancer drug target. Curr Cancer Drug Targets. 4:653–671. 2004. View Article : Google Scholar : PubMed/NCBI
13	Mishra L, Shetty K, Tang Y, Stuart A and Byers SW: The role of TGF-beta and Wnt signaling in gastrointestinal stem cells and cancer. Oncogene. 24:5775–5789. 2005. View Article : Google Scholar : PubMed/NCBI
14	Li XJ, Park ES, Park MH and Kim SM: 3,3′-Diindolylmethane suppresses the growth of gastric cancer cells via activation of the Hippo signaling pathway. Oncol Rep. 30:2419–2426. 2013.PubMed/NCBI
15	Kim SJ, Lee JS and Kim SM: 3,3′-Diindolylmethane suppresses growth of human esophageal squamous cancer cells by G1 cell cycle arrest. Oncol Rep. 27:1669–1673. 2012.PubMed/NCBI
16	Rahman KM, Li Y and Sarkar FH: Inactivation of akt and NF-kappaB play important roles during indole-3-carbinol-induced apoptosis in breast cancer cells. Nutr Cancer. 48:84–94. 2004. View Article : Google Scholar : PubMed/NCBI
17	Hong C, Kim HA, Firestone GL and Bjeldanes LF: 3,3′-Diindolylmethane (DIM) induces a G(1) cell cycle arrest in human breast cancer cells that is accompanied by Sp1-mediated activation of p21(WAF1/CIP1) expression. Carcinogenesis. 23:1297–1305. 2002. View Article : Google Scholar : PubMed/NCBI
18	Hong C, Firestone GL and Bjeldanes LF: Bcl-2 family-mediated apoptotic effects of 3,3′-diindolylmethane (DIM) in human breast cancer cells. Biochem Pharmacol. 63:1085–1097. 2002. View Article : Google Scholar : PubMed/NCBI
19	Banerjee S, Kong D, Wang Z, Bao B, Hillman GG and Sarkar FH: Attenuation of multi-targeted proliferation-linked signaling by 3,3′-diindolylmethane (DIM): From bench to clinic. Mutat Res. 728:47–66. 2011. View Article : Google Scholar : PubMed/NCBI
20	Ye S, Lee KB, Park MH, Lee JS and Kim SM: p63 regulates growth of esophageal squamous carcinoma cells via the Akt signaling pathway. Int J Oncol. 44:2153–2159. 2014.PubMed/NCBI
21	Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligo-nucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
22	Kim SM, Park YY, Park ES, Cho JY, Izzo JG, Zhang D, Kim SB, Lee JH, Bhutani MS, Swisher SG, et al: Prognostic biomarkers for esophageal adenocarcinoma identified by analysis of tumor transcriptome. PLoS One. 5:e150742010. View Article : Google Scholar : PubMed/NCBI
23	Eisen MB, Spellman PT, Brown PO and Botstein D: Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci USA. 95:14863–14868. 1998. View Article : Google Scholar : PubMed/NCBI
24	Jin H, Park MH and Kim SM: 3,3′-Diindolylmethane potentiates paclitaxel-induced antitumor effects on gastric cancer cells through the Akt/FOXM1 signaling cascade. Oncol Rep. 33:2031–2036. 2015.PubMed/NCBI
25	Kim M, Kim M, Lee S, Kuninaka S, Saya H, Lee H, Lee S and Lim DS: cAMP/PKA signalling reinforces the LATS-YAP pathway to fully suppress YAP in response to actin cytoskeletal changes. EMBO J. 32:1543–1555. 2013. View Article : Google Scholar : PubMed/NCBI
26	Cho JY, Lim JY, Cheong JH, Park YY, Yoon SL, Kim SM, Kim SB, Kim H, Hong SW, Park YN, et al: Gene expression signature-based prognostic risk score in gastric cancer. Clin Cancer Res. 17:1850–1857. 2011. View Article : Google Scholar : PubMed/NCBI
27	Heagerty PJ and Zheng Y: Survival model predictive accuracy and ROC curves. Biometrics. 61:92–105. 2005. View Article : Google Scholar : PubMed/NCBI
28	Kim SM, Leem SH, Chu IS, Park YY, Kim SC, Kim SB, Park ES, Lim JY, Heo J, Kim YJ, et al: Sixty-five gene-based risk score classifier predicts overall survival in hepatocellular carcinoma. Hepatology. 55:1443–1452. 2012. View Article : Google Scholar
29	Nicastro HL, Firestone GL and Bjeldanes LF: 3,3′-diindolyl-methane rapidly and selectively inhibits hepatocyte growth factor/c-Met signaling in breast cancer cells. J Nutr Biochem. 24:1882–1888. 2013. View Article : Google Scholar : PubMed/NCBI
30	Chen C, Chen SM, Xu B, Chen Z, Wang F, Ren J, Xu Y, Wang Y, Xiao BK and Tao ZZ: In vivo and in vitro study on the role of 3,3′-diindolylmethane in treatment and prevention of nasopharyngeal carcinoma. Carcinogenesis. 34:1815–1821. 2013. View Article : Google Scholar : PubMed/NCBI
31	Ahmad A, Ali S, Ahmed A, Ali AS, Raz A, Sakr WA and Rahman KM: 3, 3′-Diindolylmethane enhances the effectiveness of herceptin against HER-2/neu-expressing breast cancer cells. PLoS One. 8:e546572013. View Article : Google Scholar
32	Li Y, Kong D, Ahmad A, Bao B and Sarkar FH: Targeting bone remodeling by isoflavone and 3,3′-diindolylmethane in the context of prostate cancer bone metastasis. PLoS One. 7:e330112012. View Article : Google Scholar
33	Gao N, Cheng S, Budhraja A, Liu EH, Chen J, Chen D, Yang Z, Luo J, Shi X and Zhang Z: 3,3′-Diindolylmethane exhibits anti-leukemic activity in vitro and in vivo through an Akt-dependent process. PLoS One. 7:e317832012. View Article : Google Scholar
34	Weng JR, Bai LY, Chiu CF, Wang YC and Tsai MH: The dietary phytochemical 3,3′-diindolylmethane induces G2/M arrest and apoptosis in oral squamous cell carcinoma by modulating Akt-NF-κB, MAPK, and p53 signaling. Chem Biol Interact. 195:224–230. 2012. View Article : Google Scholar : PubMed/NCBI
35	Li Y, Li X and Guo B: Chemopreventive agent 3,3′-diindolyl-methane selectively induces proteasomal degradation of class I histone deacetylases. Cancer Res. 70:646–654. 2010. View Article : Google Scholar : PubMed/NCBI
36	Choi HJ, Lim Y and Park JH: Induction of G1 and G2/M cell cycle arrests by the dietary compound 3,3′-diindolylmethane in HT-29 human colon cancer cells. BMC Gastroenterol. 9:392009. View Article : Google Scholar
37	Bhatnagar N, Li X, Chen Y, Zhou X, Garrett SH and Guo B: 3,3′-diindolylmethane enhances the efficacy of butyrate in colon cancer prevention through down-regulation of survivin. Cancer Prev Res (Phila). 2:581–589. 2009. View Article : Google Scholar
38	Rahman KW, Li Y, Wang Z, Sarkar SH and Sarkar FH: Gene expression profiling revealed survivin as a target of 3,3′-diindolylmethane-induced cell growth inhibition and apoptosis in breast cancer cells. Cancer Res. 66:4952–4960. 2006. View Article : Google Scholar : PubMed/NCBI
39	Tetsu O and McCormick F: Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature. 398:422–426. 1999. View Article : Google Scholar : PubMed/NCBI
40	He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT, Morin PJ, Vogelstein B and Kinzler KW: Identification of c-MYC as a target of the APC pathway. Science. 281:1509–1512. 1998. View Article : Google Scholar : PubMed/NCBI