Cantharidin and norcantharidin inhibit the ability of MCF-7 cells to adhere to platelets via protein kinase C pathway-dependent downregulation of α2 integrin

Shou,Liu-Mei; Zhang,Qiong-Yan; Li,Wei; Xie,Xin; Chen,Kai; Lian,Lian; Li,Zhen-Yu; Gong,Fei-Ran; Dai,Ke-Sheng; Mao,Yi-Xiang; Tao,Min

doi:10.3892/or.2013.2601

Cantharidin and norcantharidin inhibit the ability of MCF-7 cells to adhere to platelets via protein kinase C pathway-dependent downregulation of α2 integrin

Authors:
- Liu-Mei Shou
- Qiong-Yan Zhang
- Wei Li
- Xin Xie
- Kai Chen
- Lian Lian
- Zhen-Yu Li
- Fei-Ran Gong
- Ke-Sheng Dai
- Yi-Xiang Mao
- Min Tao
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Affiliations: Department of Oncology, the First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China, Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536, USA, Department of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: July 8, 2013 https://doi.org/10.3892/or.2013.2601
Pages: 1059-1066
Copyright: © Shou et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Cancer metastasis is a highly coordinated and dynamic multistep process in which cancer cells interact with a variety of host cells. Morphological studies have documented the association of circulating tumor cells with host platelets, where a surface coating of platelets protects tumor cells from mechanical trauma and the immune system. Cantharidin is an active constituent of mylabris, a traditional Chinese medicine. Cantharidin and norcantharidin are potent protein phosphatase 2A (PP2A) inhibitors that exhibit in vitro and in vivo antitumor activity against several types of cancer, including breast cancer. We investigated whether cantharidin and norcantharidin could repress the ability of MCF-7 breast cancer cells to adhere to platelets. Using MTT, clone formation, apoptosis, adhesion and wound-healing assays, we found that cantharidin and norcantharidin induced apoptosis and repressed MCF-7 cell growth, adhesion and migration. Moreover, we developed a flow cytometry-based analysis of tumor cell adhesion to platelets. We proved that cantharidin and norcantharidin repressed MCF-7 cell adhesion to platelets through downregulation of α2 integrin, an adhesion molecule present on the surface of cancer cells. The repression of α2 integrin expression was found to be executed through the protein kinase C pathway, the activation of which could have been due to PP2A inhibition.

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1	Tao M, Ma D, Li Y, et al: Clinical significance of circulating tumor cells in breast cancer patients. Breast Cancer Res Treat. 129:247–254. 2011. View Article : Google Scholar : PubMed/NCBI
2	Bazou D, Santos-Martinez MJ, Medina C and Radomski MW: Elucidation of flow-mediated tumour cell-induced platelet aggregation using an ultrasound standing wave trap. Br J Pharmacol. 162:1577–1589. 2011. View Article : Google Scholar : PubMed/NCBI
3	Peng F, Wei YQ, Tian L, et al: Induction of apoptosis by norcantharidin in human colorectal carcinoma cell lines: involvement of the CD95 receptor/ligand. J Cancer Res Clin Oncol. 128:223–230. 2002. View Article : Google Scholar : PubMed/NCBI
4	Chen YN, Chen JC, Yin SC, et al: Effector mechanisms of norcantharidin-induced mitotic arrest and apoptosis in human hepatoma cells. Int J Cancer. 100:158–165. 2002. View Article : Google Scholar : PubMed/NCBI
5	Huan SK, Lee HH, Liu DZ, Wu CC and Wang CC: Cantharidin-induced cytotoxicity and cyclooxygenase 2 expression in human bladder carcinoma cell line. Toxicology. 223:136–143. 2006. View Article : Google Scholar : PubMed/NCBI
6	Williams LA, Moller W, Merisor E, Kraus W and Rosner H: In vitro anti-proliferation/cytotoxic activity of cantharidin (Spanish Fly) and related derivatives. West Indian Med J. 52:10–13. 2003.PubMed/NCBI
7	Li W, Xie L, Chen Z, et al: Cantharidin, a potent and selective PP2A inhibitor, induces an oxidative stress-independent growth inhibition of pancreatic cancer cells through G2/M cell-cycle arrest and apoptosis. Cancer Sci. 101:1226–1233. 2010. View Article : Google Scholar : PubMed/NCBI
8	Li W, Chen Z, Zong Y, et al: PP2A inhibitors induce apoptosis in pancreatic cancer cell line PANC-1 through persistent phosphorylation of IKKα and sustained activation of the NF-κB pathway. Cancer Lett. 304:117–127. 2011.PubMed/NCBI
9	Li W, Chen Z, Gong FR, et al: Growth of the pancreatic cancer cell line PANC-1 is inhibited by protein phosphatase 2A inhibitors through overactivation of the c-Jun N-terminal kinase pathway. Eur J Cancer. 47:2654–2664. 2011. View Article : Google Scholar : PubMed/NCBI
10	Carmichael J, DeGraff WG, Gazdar AF, Minna JD and Mitchell JB: Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47:936–942. 1987.PubMed/NCBI
11	Zhang G, Han J, Welch EJ, et al: Lipopolysaccharide stimulates platelet secretion and potentiates platelet aggregation via TLR4/MyD88 and the cGMP-dependent protein kinase pathway. J Immunol. 182:7997–8004. 2009. View Article : Google Scholar
12	Egan K, Crowley D, Smyth P, et al: Platelet adhesion and degranulation induce pro-survival and pro-angiogenic signalling in ovarian cancer cells. PLoS One. 6:e261252011. View Article : Google Scholar : PubMed/NCBI
13	Erpenbeck L and Schon MP: Deadly allies: the fatal interplay between platelets and metastasizing cancer cells. Blood. 115:3427–3436. 2010. View Article : Google Scholar : PubMed/NCBI
14	Honkanen RE: Cantharidin, another natural toxin that inhibits the activity of serine/threonine protein phosphatases types 1 and 2A. FEBS Lett. 330:283–286. 1993. View Article : Google Scholar : PubMed/NCBI
15	Millward TA, Zolnierowicz S and Hemmings BA: Regulation of protein kinase cascades by protein phosphatase 2A. Trends Biochem Sci. 24:186–191. 1999. View Article : Google Scholar : PubMed/NCBI
16	Schonthal AH: Role of serine/threonine protein phosphatase 2A in cancer. Cancer Lett. 170:1–13. 2001. View Article : Google Scholar : PubMed/NCBI
17	Kim YJ, Borsig L, Varki NM and Varki A: P-selectin deficiency attenuates tumor growth and metastasis. Proc Natl Acad Sci USA. 95:9325–9330. 1998. View Article : Google Scholar : PubMed/NCBI
18	Zhang N, Zhang WJ, Cai HQ, et al: Platelet adhesion and fusion to endothelial cell facilitate the metastasis of tumor cell in hypoxia-reoxygenation condition. Clin Exp Metastasis. 28:1–12. 2011. View Article : Google Scholar : PubMed/NCBI
19	Biggerstaff JP, Seth N, Amirkhosravi A, et al: Soluble fibrin augments platelet/tumor cell adherence in vitro and in vivo, and enhances experimental metastasis. Clin Exp Metastasis. 17:723–730. 1999. View Article : Google Scholar : PubMed/NCBI
20	Karpatkin S, Pearlstein E, Ambrogio C and Coller BS: Role of adhesive proteins in platelet tumor interaction in vitro and metastasis formation in vivo. J Clin Invest. 81:1012–1019. 1988. View Article : Google Scholar : PubMed/NCBI
21	Chiang HS, Yang RS and Huang TF: The Arg-Gly-Asp-containing peptide, rhodostomin, inhibits in vitro cell adhesion to extracellular matrices and platelet aggregation caused by saos-2 human osteosarcoma cells. Br J Cancer. 71:265–270. 1995. View Article : Google Scholar
22	Xu J, Zutter MM, Santoro SA and Clark RA: A three-dimensional collagen lattice activates NF-κB in human fibroblasts: role in integrin α₂gene expression and tissue remodeling. J Cell Biol. 140:709–719. 1998.PubMed/NCBI
23	Eriksson M, Arminen L, Karjalainen-Lindsberg ML and Leppa S: AP-1 regulates α2β1 integrin expression by ERK-dependent signals during megakaryocytic differentiation of K562 cells. Exp Cell Res. 304:175–186. 2005.
24	Janssens V, Goris J and Van Hoof C: PP2A: the expected tumor suppressor. Curr Opin Genet Dev. 15:34–41. 2005. View Article : Google Scholar : PubMed/NCBI