BAD overexpression inhibits cell growth and induces apoptosis via mitochondrial-dependent pathway in non-small cell lung cancer

Cancer Cell International

Weimin Li¹  Xianming Mo⁵  Yi Huang²  Na Huang³  Jing Zeng¹  Lin Mai¹  Wen Zhang¹  Bojiang Chen¹  Dan Liu¹  Man Luo¹  Li Jiang⁴ 

[1] Department of Respiratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, P.R China;Clinical Laboratory Department, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, Chengdu 610072, P.R China;Department of Respiratory Medicine, the First Affiliated Hospital of Chengdu Medical College, Chengdu 610072, P.R China;Department of Respiratory Medicine, Nanchong Central Hospital, Nanchong 637000, P.R China;Laboratory Stem Cell Biology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P.R China

关键词: Overexpression;    Cell proliferation;    Apoptosis;    Non-small cell lung cancer;    BAD protein;   

Others  :  793676
DOI  :  10.1186/1475-2867-13-53

 received in 2013-03-20, accepted in 2013-05-27,  发布年份 2013

【 摘 要 】

Background

The pro-apoptotic Bcl-2 protein BAD initiated apoptosis in human cells and has been identified as a prognostic marker in non-small cell lung cancer (NSCLC). In this study, we aimed to explore the functions of BAD in NSCLC.

Methods

Overexpression of BAD was performed by transfecting different NSCLC cell lines with wild-type BAD. Cell proliferation, cell cycle, apoptosis, and invasion were characterized in vitro. Tumorigenicity was analyzed in vivo. Western blot was performed to determine the effects of BAD overexpression on the Bcl-2 family proteins and apoptosis-related proteins.

Results

Overexpression of BAD significantly inhibited cell proliferation in H1299, H292, and SPC-A1 but not in SK-MES-1 and H460 cell lines in vitro. BAD overexpression also reduced the tumorigenicity of H1299/SPC-A1 cell in vivo. However, no appreciable effects on cell cycle distribution and invasion were observed in all these cell lines. BAD overexpression also induced apoptosis in all cell types, in which process expression of mitochondrial cytochrom c (cyto-c) and caspase 3 were increased, whereas Bcl-xl, Bcl-2, Bax and caspase 8 expressions did not changed. These findings indicated that a mitochondrial pathway, in which process cyto-c was released from mitochondrial to activate caspase 3, was involved in BAD overexpression-mediated apoptosis.

Conclusions

Our data suggested that increased expression of BAD enhance apoptosis and has negative influence on cell proliferation and tumor growth in NSCLC. Bad is a new potential target for tumor interventions.

【 授权许可】

   
2013 Jiang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Selinger CI, Cooper WA, Al-Sohaily S, Mladenova DN, Pangon L, Kennedy CW, McCaughan BC, Stirzaker C, Kohonen-Corish MR: Loss of special AT-rich binding protein 1 expression is a marker of poor survival in lung cancer. J Thorac Oncol 2011, 6:1179-1189.
• [2]Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ: Cancer statistics, 2009. CA Cancer J Clin 2009, 59:225-249.
• [3]Huang Y, Liu D, Chen B, Zeng J, Wang L, Zhang S, Mo X, Li W: Loss of Bad expression confers poor prognosis in non-small cell lung cancer. Med Oncol 2012, 29:1648-1655.
• [4]Downward J: How BAD phosphorylation is good for survival. Nat Cell Biol 1999, 1:E33-E35.
• [5]Kuroda J, Taniwaki M: Involvement of BH3-only proteins in hematologic malignancies. Crit Rev Oncol Hematol 2009, 71:89-101.
• [6]Adams JM, Cory S: The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 2007, 26:1324-1337.
• [7]Sommer P, Cowen RL, Berry A, Cookson A, Telfer BA, Williams KJ, Stratford IJ, Kay P, White A, Ray DW: Glucocorticoid receptor over-expression promotes human small cell lung cancer apoptosis in vivo and thereby slows tumor growth. Endocr Relat Cancer 2010, 17:203-213.
• [8]Ranger AM, Zha J, Harada H, Datta SR, Danial NN, Gilmore AP, Kutok JL, Le Beau MM, Greenberg ME, Korsmeyer SJ: Bad-deficient mice develop diffuse large B cell lymphoma. Proc Natl Acad Sci U S A 2003, 100:9324-9329.
• [9]Gilmore AP, Valentijn AJ, Wang P, Ranger AM, Bundred N, O’Hare MJ, Wakeling A, Korsmeyer SJ, Streuli CH: Activation of BAD by therapeutic inhibition of epidermal growth factor receptor and transactivation by insulin-like growth factor receptor. J Biol Chem 2002, 277:27643-27650.
• [10]Sinicrope FA, Rego RL, Foster NR, Thibodeau SN, Alberts SR, Windschitl HE, Sargent DJ: Proapoptotic Bad and Bid protein expression predict survival in stages II and III colon cancers. Clin Cancer Res 2008, 14:4128-4133.
• [11]Seow HF, Yip WK, Loh HW, Ithnin H, Por P, Rohaizak M: Immunohistochemical detection of phospho-Akt, phospho-BAD, HER2 and oestrogen receptors alpha and beta in Malaysian breast cancer patients. Pathol Oncol Res 2011, 16:239-248.
• [12]Huang N, Zhu J, Liu D, Li YL, Chen BJ, He YQ, Liu K, Mo XM, Li WM: Overexpression of Bcl-2-associated death inhibits A549 cell growth in vitro and in vivo. Cancer Biother Radiopharm 2012, 27:164-168.
• [13]Liu D, Huang Y, Zeng J, Chen B, Huang N, Guo N, Liu L, Xu H, Mo X, Li W: Down-regulation of JAK1 by RNA interference inhibits growth of the lung cancer cell line A549 and interferes with the PI3K/mTOR pathway. J Cancer Res Clin Oncol 2011, 137:1629-1640.
• [14]Liu D, Huang Y, Chen B, Zeng J, Guo N, Zhang S, Liu L, Xu H, Mo X, Li W: Activation of mammalian target of rapamycin pathway confers adverse outcome in nonsmall cell lung carcinoma. Cancer 2011, 117:3763-3773.
• [15]Al-Bazz YO, Underwood JC, Brown BL, Dobson PR: Prognostic significance of Akt, phospho-Akt and BAD expression in primary breast cancer. Eur J Cancer 2009, 45:694-704.
• [16]Marchion DC, Cottrill HM, Xiong Y, Chen N, Bicaku E, Fulp WJ, Bansal N, Chon HS, Stickles XB, Kamath SG, Hakam A, Li L, Su D, Moreno C, Judson PL, Berchuck A, Wenham RM, Apte SM, Gonzalez-Bosquet J, Bloom GC, Eschrich SA, Sebti S, Chen DT, Lancaster JM, et al.: BAD phosphorylation determines ovarian cancer chemosensitivity and patient survival. Clin Cancer Res 2012, 17:6356-6366.
• [17]Danial NN: BAD: undertaker by night, candyman by day. Oncogene 2008, 27(Suppl 1):S53-S70.
• [18]Mok CL, Gil-Gómez G, Williams O, Coles M, Taga S, Tolaini M, Norton T, Kioussis D, Brady HJ: Bad can act as a key regulator of T cell apoptosis and T cell development. J Exp Med 1999, 189:575-586.
• [19]Taghiyev AF, Guseva NV, Harada H, Knudson CM, Rokhlin OW, Cohen MB: Overexpression of BAD potentiates sensitivity to tumor necrosis factor-related apoptosis-inducing ligand treatment in the prostatic carcinoma cell line LNCaP. Mol Cancer Res 2003, 1:500-507.
• [20]Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ: Bad, a heterodimeric partner for Bcl-XL and Bcl-2, displaces Bax and promotes cell death. Cell 1995, 80:285-291.
• [21]Fernando R, Foster JS, Bible A, Strom A, Pestell RG, Rao M, Saxton A, Baek SJ, Yamaguchi K, Donnell R, et al.: Breast cancer cell proliferation is inhibited by BAD: regulation of cyclin D1. J Biol Chem 2007, 282:28864-28873.
• [22]Lee KW, Kim SG, Kim HP, Kwon E, You J, Choi HJ, Park JH, Kang BC, Im SA, Kim TY, Kim WH, Bang YJ, et al.: Enzastaurin, a protein kinase C beta inhibitor, suppresses signaling through the ribosomal S6 kinase and bad pathways and induces apoptosis in human gastric cancer cells. Cancer Res 2008, 68:1916-1926.
• [23]Hengartner MO: The biochemistry of apoptosis. Nature 2000, 407:770-776.
• [24]Chauhan D, Pandey P, Ogata A, Teoh G, Krett N, Halgren R, Rosen S, Kufe D, Kharbanda S, Anderson K: Cytochrome c-dependent and -independent induction of apoptosis in multiple myeloma cells. J Biol Chem 1997, 272:29995-29997.
• [25]Cheng EH, Wei MC, Weiler S, Flavell RA, Mak TW, Lindsten T, Korsmeyer SJ: BCL-2, BCL-X(L) sequester BH3 domain-only molecules preventing BAX- and BAK-mediated mitochondrial apoptosis. Mol Cell 2001, 8:705-711.
• [26]Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB, Korsmeyer SJ: Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001, 292:727-730.
• [27]Xiao D, Singh SV: Diallyl trisulfide, a constituent of processed garlic, inactivates Akt to trigger mitochondrial translocation of BAD and caspase-mediated apoptosis in human prostate cancer cells. Carcinogenesis 2006, 27:533-540.
• [28]Smith AJ, Karpova Y, D’Agostino R Jr, Willingham M, Kulik G: Expression of the Bcl-2 protein BAD promotes prostate cancer growth. PLoS One 2009, 4:e6224.
• [29]Polzien L, Baljuls A, Albrecht M, Hekman M, Rapp UR: BAD contributes to RAF-mediated proliferation and cooperates with B-RAF-V600E in cancer signaling. J Biol Chem 2011, 286:17934-17944.
• [30]Maslyar DJ, Aoki M, Vogt PK: The growth-promoting activity of the Bad protein in chicken embryo fibroblasts requires binding to protein 14-3-3. Oncogene 2001, 20:5087-5092.
• [31]Chattopadhyay A, Chiang CW, Yang E: BAD/BCL-[X(L)] heterodimerization leads to bypass of G0/G1 arrest. Oncogene 2001, 20:4507-4518.
• [32]Cekanova M, Fernando R, Wimalasena J: P1-02-11: the BCL2 antagonist of death, BAD is down-regulated in breast cancer and inhibits cancer cell invasion. Abstracts: Thirty-Fourth Annual CTRC-AACR San Antonio Breast Cancer Symposium; 2011.