Ardipusilloside I induces apoptosis by regulating Bcl-2 family proteins in human mucoepidermoid carcinoma Mc3 cells

BMC Complementary and Alternative Medicine

Xiao-Juan Wang³  Peng-Yuan Wang³  Wei-Dong Zhang³  Xin Xiao³  Rong Wang³  Song Jin²  Tao-Li Zhang³  Xiao-Fang Xu¹ 

[1] Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, 145 Chang Le Xi Road, Xi’an, Shaanxi Province 710032, China;Department of Oral and Maxillofacial Surgery, The 22nd hospital of PLA, Middle-Yanqiao Road, Ge’ermu, Qinghai Province 816000, China;Department of Pharmaceutical Preparation, School of Stomatology, The Fourth Military Medical University, 145 Chang Le Xi Road, Xi’an, Shaanxi Province 710032, China

关键词: Ardipusilloside I;    Traditional Chinese herb;    Bcl-2;    Apoptosis;    Mc3 cell line;    Mucoepidermoid carcinoma;   

Others  :  1220518
DOI  :  10.1186/1472-6882-13-322

 received in 2012-05-17, accepted in 2013-11-15,  发布年份 2013

【 摘 要 】

Background

Ardisia pusilla A. DC., family Myrsinaceae, is a traditional Chinese medicine named Jiu Jie Long with a variety of pharmacological functions including anti-cancer activities. In this study, we purified a natural triterpenoid saponin, ardipusilloside I, from Ardisia pusilla, and show that it exhibits inhibitory activities in human mucoepidermoid carcinoma Mc3 cells. We also investigated the underlying mechanisms of proliferation inhibition that ardipusilloside I exerts on Mc3 cells.

Methods

MTT test was used to detect cell proliferation. Cell apoptosis was detected by transmission electron microscopy, Hoechst-33342 staining, DNA fragmentation detection, and flow cytometry. We also used western blot analysis to detect the potential mechanisms of apoptosis.

Results

Ardipusilloside I affected the viability of Mc3 cells in a dose- and time-dependent manner. The IC50 of ardipusilloside I was approximately 9.98 μg/ml at 48 h of treatment. Characteristic morphological changes of apoptosis, including nuclear condensation, boundary aggregation and splitting, and DNA fragmentation, were seen after treatment with 10 μg/ml ardipusilloside I for 48 h. Western blots demonstrated that ardipusilloside I caused Mc3 cell death through the induction of apoptosis by downregulation of Bcl-2 protein levels and upregulation of Bax and caspase-3 protein levels.

Conclusions

Our results revealed that ardipusilloside I could be a new active substance for mucoepidermoid carcinoma treatment. We demonstrated that the potential mechanism of inhibition might be through the induction of apoptosis by regulation of Bcl-2 family protein levels. This suggests a further rationale for the development of ardipusilloside I as an anti-cancer agent.

【 授权许可】

   
2013 Xu et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Pires FR, Pringle GA, de Almeida OP, Chen SY: Intra-oral minor salivary gland tumors: a clinicopathological study of 546 cases. Oral Oncol 2007, 43(5):463-470.
• [2]Kokemueller H, Brueggemann N, Swennen G, Eckardt A: Mucoepidermoid carcinoma of the salivary glands–clinical review of 42 cases. Oral Oncol 2005, 41(1):3-10.
• [3]O’Neill ID: t(11;19) translocation and CRTC1-MAML2 fusion oncogene in mucoepidermoid carcinoma. Oral Oncol 2009, 45(1):2-9.
• [4]Vattemi E, Graiff C, Sava T, Pedersini R, Caldara A, Mandara M: Systemic therapies for recurrent and/or metastatic salivary gland cancers. Expert Rev Anticancer Ther 2008, 8(3):393-402.
• [5]Xu XF, Cai BL, Guan SM, Li Y, Wu JZ, Wang Y, Liu B: Baicalin induces human mucoepidermoid carcinoma Mc3 cells apoptosis in vitro and in vivo. Invest New Drugs 2011, 29(4):637-645.
• [6]Zhang T, Wang X, Wang R, Gu Y, Wang R: Zhang W: [Inhibition of the proliferation ofhuman mucoepidermoid carcinoma Mc3 cells by Ardipusilloside I]. J Pract Stomatol 2011, 19(7):264-270.
• [7]Zhang QH, Huang SL, Wang XJ: [Studies on the sapogenins and prosapogenins in Ardisia pusilla A. DC]. Zhongguo Zhong Yao Za Zh 1993, 18(9):545-547. 574
• [8]Xiong J, Cheng G, Tang H, Zhen H, Zhang X: Ardipusilloside I induces apoptosis in human glioblastoma cells through a caspase-8-independent FasL/Fas-signaling pathway. Environ Toxicol Phar 2009, 27(2):264-270.
• [9]Zhang Y, Qu Y, Zhang J, Wang X: Ardipusilloside I purified from Ardisia pusilla competitively binds VEGFR and induces apoptosis in NCI-H460 cells. Phytomedicine 2010, 17(7):519-526.
• [10]Lou L, Ye W, Chen Y, Wu S, Jin L, He J, Tao X, Zhu J, Chen X, Deng A, et al.: Ardipusilloside inhibits survival, invasion and metastasis of human hepatocellular carcinoma cells. Phytomedicine 2012, 19(7):603-608.
• [11]Wen DS, Zhu XL, Guan SM, Wu YM, Yu LL, Wu JZ: Silencing of CXCR4 inhibits the proliferation, adhesion, chemotaxis and invasion of salivary gland mucoepidermoid carcinoma Mc3 cells in vitro. Oral Oncol 2008, 44(6):545-554.
• [12]Chen MH, Chen RS, Hsu YH, Chen YJ, Young TH: Proliferation and phenotypic preservation of rat parotid acinar cells. Tissue Eng 2005, 11(3–4):526-534.
• [13]Bajbouj K, Schulze-Luehrmann J, Diermeier S, Amin A, Schneider-Stock R: The anticancer effect of saffron in two p53 isogenic colorectal cancer cell lines. BMC Complement Altern Med 2012, 12(1):69. BioMed Central Full Text
• [14]Swellam T, Miyanaga N, Onozawa M, Hattori K, Kawai K, Shimazui T, Akaza H: Antineoplastic activity of honey in an experimental bladder cancer implantation model: in vivo and in vitro studies. Int J Urol 2003, 10(4):213-219.
• [15]Jegham H, Roy J, Maltais R, Desnoyers S, Poirier D: A novel aminosteroid of the 5alpha-androstane-3alpha,17beta-diol family induces cell cycle arrest and apoptosis in human promyelocytic leukemia HL-60 cells. Invest New Drugs 2012, 30(1):176-185.
• [16]Xue X, Qu XJ, Gao ZH, Sun CC, Liu HP, Zhao CR, Cheng YN, Lou HX: Riccardin D, a novel macrocyclic bisbibenzyl, induces apoptosis of human leukemia cells by targeting DNA topoisomerase II. Invest New Drugs 2012, 30(1):212-222.
• [17]Tao X, Wang P, Yang X, Yao H, Liu J, Cao Y: [Inhibitory effect of ardipusilloside-I on Lewis pulmonary carcinoma and hepatocarcinoma SMMC-7721]. Zhong Yao Cai 2005, 28(7):574-577.
• [18]Li HL, Ye KH, Zhang HW, Luo YR, Ren XD, Xiong AH, Situ R: Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells. Cell Res 2001, 11(4):311-315.
• [19]Hanahan D, Weinberg RA: The hallmarks of cancer. Cell 2000, 100(1):57-70.
• [20]Snider BJ, Gottron FJ, Choi DW: Apoptosis and necrosis in cerebrovascular disease. Ann N Y Acad Sci 1999, 893:243-253.
• [21]Klein S, McCormick F, Levitzki A: Killing time for cancer cells. Nat Rev Cancer 2005, 5(7):573-580.
• [22]Marzo I, Naval J: Bcl-2 family members as molecular targets in cancer therapy. Biochem Pharmacol 2008, 76(8):939-946.
• [23]Tsujimoto Y, Croce CM: Analysis of the structure, transcripts, and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci U S A 1986, 83(14):5214-5218.
• [24]Reed JC: Double identity for proteins of the Bcl-2 family. Nature 1997, 387(6635):773-776.
• [25]Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, Ody C, Donati Y, Barazzone AC: Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway. Free Radic Biol Med 2007, 42(7):1062-1074.
• [26]Bernecker OY, Huq F, Heist EK, Podesser BK, Hajjar RJ: Apoptosis in heart failure and the senescent heart. Cardiovasc Toxicol 2003, 3(3):183-190.
• [27]Tsujimoto Y, Finger LR, Yunis J, Nowell PC, Croce CM: Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science 1984, 226(4678):1097-1099.
• [28]Kitada S, Pedersen IM, Schimmer AD, Reed JC: Dysregulation of apoptosis genes in hematopoietic malignancies. Oncogene 2002, 21(21):3459-3474.
• [29]Rampino N, Yamamoto H, Ionov Y, Li Y, Sawai H, Reed JC, Perucho M: Somatic frameshift mutations in the BAX gene in colon cancers of the microsatellite mutator phenotype. Science 1997, 275(5302):967-969.
• [30]Liu J, Wu Y, Wang B, Yuan X, Fang B: High levels of glucose induced the caspase-3/PARP signaling pathway, leading to apoptosis in human periodontal ligament fibroblasts. Cell Biochem Biophys 2012. (Epub DOI: 10.1007/s12013-012-9470-y)
• [31]Calandria C, Irurzun A, Barco A, Carrasco L: Individual expression of poliovirus 2Apro and 3Cpro induces activation of caspase-3 and PARP cleavage in HeLa cells. Virus Res 2004, 104(1):39-49.