Analgesic-antitumor peptide induces apoptosis and inhibits the proliferation of SW480 human colon cancer cells

Gu,Yu; Liu,Shen-Lin; Ju,Wen-Zheng; Li,Chang-Yin; Cao,Peng

doi:10.3892/ol.2012.1049

Analgesic-antitumor peptide induces apoptosis and inhibits the proliferation of SW480 human colon cancer cells

Authors:
- Yu Gu
- Shen-Lin Liu
- Wen-Zheng Ju
- Chang-Yin Li
- Peng Cao
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Affiliations: The First Clinical Medical College, Nanjing University of Traditional Medicine, Nanjing, Jiangsu, P.R. China, Laboratory of Clinical Pharmacokinetics of Traditional Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, P.R. China, Laboratory of Cellular and Molecular Biology, Jiangsu Province Institute of Traditional Chinese Medicine, Nanjing, Jiangsu, P.R. China
Published online on: November 27, 2012 https://doi.org/10.3892/ol.2012.1049
Pages: 483-488

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Abstract

Colorectal cancer is one of the most common malignant tumors, and is associated with significant morbidity and mortality. In this study, recombinant analgesic-antitumor peptide (rAGAP), a protein consisting of small ubiquitin-related modifier (SUMO) linked with a hexa-histidine tag, was used as an antitumor analgesic peptide. The purpose of the present study was to investigate the antitumor activity of rAGAP in human colon adenocarcinoma SW480 cells and its potential molecular mechanisms of action. In this study, cell viability and apoptosis of rAGAP-treated SW480 cells was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry and 4',6-diamidino-2-phenylindole (DAPI) staining. Western blotting was used to investigate the effects of rAGAP on p27, Bcl-2/Bax and PTEN/PI3K/Akt cellular signal transduction. Our results showed that rAGAP not only enhanced apoptosis, but also inhibited the proliferation of SW480 cells. rAGAP upregulates the expression of p27 in SW480 cells and leads to cell cycle arrest in the G1 phase. Furthermore, rAGAP significantly increases the production of Bax and PTEN and suppresses the activation of Bcl-2, phosphatidylinositol 3-kinase (PI3K) and phospho-Akt (p-Akt) in SW480 cells. These results suggest that rAGAP may be a potential new anti-colorectal cancer drug.

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1.	Center MM, Jemal A, Smith RA and Ward E: Worldwide variations in colorectal cancer. CA Cancer J Clin. 59:366–378. 2009. View Article : Google Scholar : PubMed/NCBI
2.	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
3.	Weekes J, Lam AK, Sebesan S and Ho YH: Irinotecan therapy and molecular targets in colorectal cancer: a systemic review. World J Gastroenterol. 15:3597–3602. 2009. View Article : Google Scholar : PubMed/NCBI
4.	Zhao Y, Cai X, Ye T, et al: Analgesic-antitumor peptide inhibits proliferation and migration of SHG-44 human malignant glioma cells. J Cell Biochem. 112:2424–2434. 2011. View Article : Google Scholar : PubMed/NCBI
5.	Cao P, Yu J, Lu W, et al: Expression and purification of an antitumor-analgesic peptide from the venom of Mesobuthus martensii Karsch by small ubiquitin-related modifier fusion in Escherichia coli. Biotechnol Prog. 26:1240–1244. 2010. View Article : Google Scholar : PubMed/NCBI
6.	Guo Y, Srinivasula SM, Druilhe A, Fernandes-Alnemri T and Alnemri ES: Caspase-2 induces apoptosis by releasing proapoptotic proteins from mitochondria. J Biol Chem. 277:13430–13437. 2002. View Article : Google Scholar : PubMed/NCBI
7.	Amado RG, Wolf M, Peeters M, et al: Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J Clin Oncol. 26:1626–1634. 2008. View Article : Google Scholar : PubMed/NCBI
8.	Karapetis CS, Khambata-Ford S, Jonker DJ, et al: K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med. 359:1757–1765. 2008. View Article : Google Scholar : PubMed/NCBI
9.	Deorukhkar A, Krishnan S, Sethi G and Aggarwal BB: Back to basics: how natural products can provide the basis for new therapeutics. Expert Opin Investig Drugs. 16:1753–1773. 2007. View Article : Google Scholar : PubMed/NCBI
10.	Newman DJ and Cragg GM: Natural products as sources of new drugs over the last 25 years. J Nat Prod. 70:461–477. 2007.PubMed/NCBI
11.	du Plessis LH, Elgar D and du Plessis JL: Southern African scorpion toxins: an overview. Toxicon. 51:1–9. 2008.PubMed/NCBI
12.	Rodriguez de la Vega RC and Possani LD: Overview of scorpion toxins specific for Na+ channels and related peptides: biodiversity, structure-function relationships and evolution. Toxicon. 46:831–844. 2005.PubMed/NCBI
13.	Fan S, Sun Z, Jiang D, et al: BmKCT toxin inhibits glioma proliferation and tumor metastasis. Cancer Lett. 291:158–166. 2010. View Article : Google Scholar : PubMed/NCBI
14.	Das Gupta S, Debnath A, Saha A, et al: Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leukemia. 31:817–825. 2007.
15.	Gao R, Zhang Y and Gopalakrishnakone P: Purification and N-terminal sequence of a serine proteinase-like protein (BMK-CBP) from the venom of the Chinese scorpion (Buthus martensii Karsch). Toxicon. 52:348–353. 2008. View Article : Google Scholar : PubMed/NCBI
16.	Chaudhry MA: Base excision repair of ionizing radiation-induced DNA damage in G1 and G2 cell cycle phases. Cancer Cell Int. 7:152007. View Article : Google Scholar : PubMed/NCBI
17.	Sitko JC, Yeh B, Kim M, et al: SOCS3 regulates p21 expression and cell cycle arrest in response to DNA damage. Cell Signal. 20:2221–2230. 2008. View Article : Google Scholar : PubMed/NCBI
18.	Narbonne-Reveau K and Lilly M: The Cyclin-dependent kinase inhibitor Dacapo promotes genomic stability during premeiotic S phase. Mol Biol Cell. 20:1960–1969. 2009. View Article : Google Scholar : PubMed/NCBI
19.	Liu S and Yamauchi H: p27-Associated G1 arrest induced by hinokitiol in human malignant melanoma cells is mediated via down-regulation of pRb, Skp2 ubiquitin ligase, and impairment of Cdk2 function. Cancer Lett. 286:240–249. 2009. View Article : Google Scholar : PubMed/NCBI
20.	Connor MK, Kotchetkov R, Cariou S, et al: CRM1/Ran-mediated nuclear export of p27(Kip1) involves a nuclear export signal and links p27 export and proteolysis. Mol Biol Cell. 14:201–213. 2003. View Article : Google Scholar : PubMed/NCBI
21.	Yakes FM, Chinratanalab W, Ritter CA, King W, Seelig S and Arteaga CL: Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt is required for antibody-mediated effects on p27, cyclin D1, and antitumor action. Cancer Res. 62:4132–4141. 2002.PubMed/NCBI
22.	Kerkhoff E, Simpson JC, Leberfinger CB, et al: The Spir actin organizers are involved in vesicle transport processes. Curr Biol. 11:1963–1968. 2001. View Article : Google Scholar : PubMed/NCBI
23.	Jones SM and Kazlauskas A: Growth-factor-dependent mitogenesis requires two distinct phases of signalling. Nat Cell Biol. 3:165–172. 2001. View Article : Google Scholar : PubMed/NCBI
24.	Liang J, Zubovitz J, Petrocelli T, et al: PKB/Akt phosphorylates p27, impairs nuclear import of p27 and opposes p27-mediated G1 arrest. Nat Med. 8:1153–1160. 2002. View Article : Google Scholar : PubMed/NCBI
25.	Liang J and Slingerland JM: Multiple roles of the PI3K/PKB (Akt) pathway in cell cycle progression. Cell Cycle. 2:339–345. 2003. View Article : Google Scholar : PubMed/NCBI
26.	Petros AM, Olejniczak ET and Fesik SW: Structural biology of the Bcl-2 family of proteins. Biochim Biophys Acta. 1644:83–94. 2004. View Article : Google Scholar : PubMed/NCBI
27.	Chowdhury I, Tharakan B and Bhat GK: Caspases - an update. Comp Biochem Physiol B Biochem Mol Biol. 151:10–27. 2008. View Article : Google Scholar : PubMed/NCBI
28.	D'Amelio M, Tino E and Cecconi F: The apoptosome: emerging insights and new potential targets for drug design. Pharm Res. 25:740–751. 2008.PubMed/NCBI
29.	Vivanco I and Sawyers CL: The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer. 2:489–501. 2002. View Article : Google Scholar : PubMed/NCBI
30.	Di Cristofano A and Pandolfi PP: The multiple roles of PTEN in tumor suppression. Cell. 100:387–390. 2000.PubMed/NCBI
31.	Laurent-Puig P, Cayre A, Manceau G, et al: Analysis of PTEN, BRAF, and EGFR status in determining benefit from cetuximab therapy in wild-type KRAS metastatic colon cancer. J Clin Oncol. 27:5924–5930. 2009. View Article : Google Scholar : PubMed/NCBI
32.	Loupakis F, Pollina L, Stasi I, et al: PTEN expression and KRAS mutations on primary tumors and metastases in the prediction of benefit from cetuximab plus irinotecan for patients with metastatic colorectal cancer. J Clin Oncol. 27:2622–2629. 2009. View Article : Google Scholar : PubMed/NCBI
33.	Frattini M, Saletti P, Romagnani E, et al: PTEN loss of expression predicts cetuximab efficacy in metastatic colorectal cancer patients. Br J Cancer. 97:1139–1145. 2007. View Article : Google Scholar : PubMed/NCBI
34.	Vyas S, Juin P, Hancock D, et al: Differentiation-dependent sensitivity to apoptogenic factors in PC12 cells. J Biol Chem. 279:30983–30993. 2004. View Article : Google Scholar : PubMed/NCBI
35.	Majewski N, Nogueira V, Robey RB and Hay N: Akt inhibits apoptosis downstream of BID cleavage via a glucose-dependent mechanism involving mitochondrial hexokinases. Mol Cell Biol. 24:730–740. 2004. View Article : Google Scholar
36.	Engelman JA, Luo J and Cantley LC: The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet. 7:606–619. 2006. View Article : Google Scholar : PubMed/NCBI
37.	Carnero A: The PKB/AKT pathway in cancer. Curr Pharm Des. 16:34–44. 2010. View Article : Google Scholar : PubMed/NCBI
38.	Song G, Ouyang G and Bao S: The activation of Akt/PKB signaling pathway and cell survival. J Cell Mol Med. 9:59–71. 2005. View Article : Google Scholar : PubMed/NCBI
39.	Liu J, Yin S, Reddy N, Spencer C and Sheng S: Bax mediates the apoptosis-sensitizing effect of maspin. Cancer Res. 64:1703–1711. 2004. View Article : Google Scholar : PubMed/NCBI
40.	Maianski NA, Geissler J, Srinivasula SM, Alnemri ES, Roos D and Kuijpers TW: Functional characterization of mitochondria in neutrophils: a role restricted to apoptosis. Cell Death Differ. 11:143–153. 2004. View Article : Google Scholar : PubMed/NCBI
41.	Kang MH and Reynolds CP: Bcl-2 inhibitors: targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res. 15:1126–1132. 2009. View Article : Google Scholar : PubMed/NCBI