Colchicine induces apoptosis in HT‑29 human colon cancer cells via the AKT and c-Jun N-terminal kinase signaling pathways

Huang,Zhen; Xu,Ye; Peng,Wei

doi:10.3892/mmr.2015.4222

Colchicine induces apoptosis in HT‑29 human colon cancer cells via the AKT and c-Jun N-terminal kinase signaling pathways

Authors:
- Zhen Huang
- Ye Xu
- Wei Peng
View Affiliations

Affiliations: Department of General Surgery, Shanghai Traditional Chinese Medicine‑Integrated Hospital, Shanghai 200082, P.R. China
Published online on: August 12, 2015 https://doi.org/10.3892/mmr.2015.4222
Pages: 5939-5944

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Colchicine is a natural compound, which belongs to the botanical family Colchicaceae and prevents growth of cancer cells via antimitotic activity by interacting with microtubules. Although numerous studies have demonstrated that the effect of colchicine on cell apoptosis is mediated by the activation of caspase‑3, the signaling pathways involved in the process remain unknown. In the current study, evidence is presented regarding the missing information using HT‑29 human colon cancer cells. The effect of colchicine on apoptosis in HT‑29 cells and the apoptosis‑associated signaling pathways were determined using various methods, including cell viability assay, Annexin V/propidium idodide (PI) binding, PI staining, Hoechst 33342 staining, mitochondrial membrane potential (Δψm) assay, reactive oxygen species (ROS) assay and western blot analysis. Colchicine was observed to induce a dose‑dependent reduction in cell viability in HT‑29 cells and early apoptosis occurred when the cells were treated with 1 µg/ml colchicine. Furthermore, colchicine treatment induced a loss of Δψm, increased ROS production, activated caspase‑3, upregulated BAX expression and downregulated Bcl‑2 expression, which evidenced the colchicine activity on apoptosis, potentially by acting via the intrinsic apoptotic signaling pathway. Colchicine increased phosphorylation of p38, although not phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase, which indicates that colchicine activates the p38 signaling pathway in order to induce cell apoptosis. Therefore, colchicine exhibited significant growth inhibition of the HT‑29 colon cancer cell line and induced apoptosis in the cells via the mitochondrial pathway, which is regulated by p38 signaling pathways.

View Figures

View References

1	Giovannucci E: Modifiable risk factors for colon cancer. Gastroenterol Clin North Am. 31:925–943. 2002. View Article : Google Scholar : PubMed/NCBI
2	Couzin J: Cancer T cells a boon for colon cancer prognosis. Science. 313:1868–1869. 2006. View Article : Google Scholar : PubMed/NCBI
3	François Y and Vignal J: Cancer of the colon. Etiology, physiopathology, diagnosis, development and prognosis, principles of the surgical treatment. Rev Prat. 41:1221–1225. 1991.
4	Chiu BC, Ji BT, Dai Q, Gridley G, McLaughlin JK, Gao YT, Fraumeni JF Jr and Chow WH: Dietary factors and risk of colon cancer in Shanghai, China. Cancer Epidemiol, Biomarkers Prev. 12:201–208. 2003.
5	Barrier A, Boelle PY, Roser F, Gregg J, Tse C, Brault D, Lacaine F, Houry S, Huguier M, Franc B, et al: Stage II colon cancer prognosis prediction by tumor gene expression profiling. J Clin Oncol. 24:4685–4691. 2006. View Article : Google Scholar : PubMed/NCBI
6	Jee SH, Moon SM, Shin US, Yang HM and Hwang DY: Effectiveness of adjuvant chemotherapy with 5-FU/leucovorin and prognosis in stage II colon cancer. J Korean Soc Coloproctol. 27:322–328. 2011. View Article : Google Scholar
7	Dotan E and Cohen SJ: Challenges in the management of stage II colon cancer. Semin Oncol. 38:511–520. 2011. View Article : Google Scholar : PubMed/NCBI
8	Risinger AL and Mooberry SL: Taccalonolides: Novel micro-tubule stabilizers with clinical potential. Cancer Lett. 291:14–19. 2010. View Article : Google Scholar :
9	Sivakumar G: Colchicine semisynthetics: Chemotherapeutics for cancer? Curr Med Chem. 20:892–898. 2013.
10	Risinger AL, Giles FJ and Mooberry SL: Microtubule dynamics as a target in oncology. Cancer Treat Rev. 35:255–261. 2009. View Article : Google Scholar : PubMed/NCBI
11	Jordan MA and Wilson L: Microtubules as a target for anticancer drugs. Nat Rev Cancer. 4:253–265. 2004. View Article : Google Scholar : PubMed/NCBI
12	Ravelli RB, Gigant B, Curmi PA, Jourdain I, Lachkar S, Sobel A and Knossow M: Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain. Nature. 428:198–202. 2004. View Article : Google Scholar : PubMed/NCBI
13	Bhattacharyya B, Panda D, Gupta S and Banerjee M: Anti-mitotic activity of colchicine and the structural basis for its interaction with tubulin. Med Res Rev. 28:155–183. 2008. View Article : Google Scholar
14	Chopra A, Anderson A and Giardina C: Novel piperazine-based compounds inhibit microtubule dynamics and sensitize colon cancer cells to tumor necrosis factor-induced apoptosis. J Biol Chem. 289:2978–2991. 2014. View Article : Google Scholar :
15	Acharya BR, Chatterjee A, Ganguli A, Bhattacharya S and Chakrabarti G: Thymoquinone inhibits microtubule polymerization by tubulin binding and causes mitotic arrest following apoptosis in A549 cells. Biochimie. 97:78–91. 2014. View Article : Google Scholar
16	Rai A, Gupta TK, Kini S, Kunwar A, Surolia A and Panda D: CXI-benzo-84 reversibly binds to tubulin at colchicine site and induces apoptosis in cancer cells. Biochem Pharmacol. 86:378–391. 2013. View Article : Google Scholar : PubMed/NCBI
17	Magalhães HI, Wilke DV, Bezerra DP, Cavalcanti BC, Rotta R, de Lima DP, Beatriz A, Moraes MO, Diniz-Filho J and Pessoa C: (4-Methoxyphenyl)(3,4,5-trimethoxyphenyl)methanone inhibits tubulin polymerization, induces G2/M arrest and triggers apoptosis in human leukemia HL-60 cells. Toxicol Appl Pharmacol. 272:117–126. 2013. View Article : Google Scholar
18	Chiang NJ, Lin CI, Liou JP, Kuo CC, Chang CY, Chen LT and Chang JY: A novel synthetic microtubule inhibitor, MPT0B214 exhibits antitumor activity in human tumor cells through mitochondria-dependent intrinsic pathway. PLoS One. 8:e589532013. View Article : Google Scholar : PubMed/NCBI
19	Wu J, Yi W, Jin L, Hu D and Song B: Antiproliferative and cell apoptosis-inducing activities of compounds from Buddleja davidii in Mgc-803 cells. Cell Div. 7:202012. View Article : Google Scholar : PubMed/NCBI
20	Wang X: The expanding role of mitochondria in apoptosis. Genes Dev. 15:2922–2933. 2001.PubMed/NCBI
21	Zamzami N, Marchetti P, Castedo M, Hirsch T, Susin SA, Masse B and Kroemer G: Inhibitors of permeability transition interfere with the disruption of the mitochondrial trans-membrane potential during apoptosis. FEBS Lett. 384:53–57. 1996. View Article : Google Scholar : PubMed/NCBI
22	Yan SL, Huang CY, Wu ST and Yin MC: Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines. Toxicol In Vitro. 24:842–848. 2010. View Article : Google Scholar
23	Roux PP and Blenis J: ERK and p38 MAPK-activated protein kinases: A family of protein kinases with diverse biological functions. Microbiol Mol Biol Rev. 68:320–344. 2004. View Article : Google Scholar : PubMed/NCBI
24	Tsuruta F, Sunayama J, Mori Y, Hattori S, Shimizu S, Tsujimoto Y, Yoshioka K, Masuyama N and Gotoh Y: JNK promotes Bax translocation to mitochondria through phosphorylation of 14-3-3 proteins. EMBO J. 23:1889–1899. 2004. View Article : Google Scholar : PubMed/NCBI
25	Pugazhenthi S, Nesterova A, Sable C, Heidenreich KA, Boxer LM, Heasley LE and Reusch JE: Akt/protein kinase B up-regulates Bcl-2 expression through cAMP-response element-binding protein. J Biol Chem. 275:10761–10766. 2000. View Article : Google Scholar