Prokaryotically and eukaryotically expressed interleukin-24 induces breast cancer growth suppression via activation of apoptosis and inhibition of tumor angiogenesis

Wei,Shaohua; Cao,Hua; Zhou,Xiaoyan; Wu,Haorong; Yang,Jicheng

doi:10.3892/mmr.2014.3136

Prokaryotically and eukaryotically expressed interleukin-24 induces breast cancer growth suppression via activation of apoptosis and inhibition of tumor angiogenesis

Authors:
- Shaohua Wei
- Hua Cao
- Xiaoyan Zhou
- Haorong Wu
- Jicheng Yang
View Affiliations

Affiliations: Department of General Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China, Department of Cell and Molecular Biology, College of Medicine, Soochow University, Suzhou, Jiangsu 215123, P.R. China
Published online on: December 24, 2014 https://doi.org/10.3892/mmr.2014.3136
Pages: 3673-3681

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

Melanoma differentiation‑associated‑7 (mda‑7)/interleukin‑24 (IL‑24), a unique cytokine‑tumor suppressor, exerts tumor‑selective killing activity in numerous types of cancer cell. Although eukaryotically and prokaryotically expressed recombinant human (rh)IL‑24 proteins have been previously shown to produce potent antitumor effects, to the best of our knowledge, no side‑by‑side study has been conducted that compares the two proteins directly. In the present study, rhIL‑24 protein was expressed in BL21 Escherichia coli transformed with the pET‑21a(+)‑hIL‑24 plasmid by isopropyl‑β‑D‑1‑thiogalactopyranoside induction. Following a denaturing and renaturing process, the soluble rhIL‑24 was purified using a Q‑Sepharose column. rhIL‑24 protein was also expressed in Chinese hamster ovary mammalian cells stably transfected with the pcDNA3‑hIL‑24 plasmid. The in vitro antitumor efficacies of the two treatments were compared using the MDA‑MB‑231 human breast cancer cell line. Furthermore, the therapeutic efficacies of the bacteria‑derived rhIL‑24 protein and the liposome‑coated pcDNA3‑hIL‑24 naked plasmid were evaluated in athymic nude mice with subcutaneously xenografted MDA‑MB‑231 cell tumors. The prokaryotically expressed/purified rhIL‑24 protein and the eukaryotically expressed rhIL‑24 in the cell supernate were revealed to be capable of efficiently suppressing MDA‑MB‑231 tumor growth in vitro. Similarly, the administration of bacteria‑derived rhIL‑24 protein and pcDNA3‑hIL‑24 naked plasmid also provided therapeutic benefits in the treatment of in vivo MDA‑MB‑231 xenografted tumors. The retarded in vitro and in vivo breast cancer growth elicited by rhIL‑24 was closely associated with the upregulation of the ratio of anti‑apoptotic B cell lymphoma 2 (Bcl‑2) to pro‑apoptotic Bcl‑2‑associated X protein (Bax), as well as the activation of caspase‑3 followed by marked induction of apoptosis, and the notable inhibition of tumor angiogenesis. Thus, the results of the present study indicate that prokaryotically expressed rhIL‑24 protein may be an alternate and promising antitumor agent in human breast cancer or other types of cancer.

View Figures

View References

1	Sauane M, Gopalkrishnan RV, Sarkar D, Su ZZ, Lebedeva IV, Dent P, et al: MDA-7/IL-24: novel cancer growth suppressing and apoptosis inducing cytokine. Cytokine Growth Factor Rev. 14:35–51. 2003. View Article : Google Scholar
2	Fisher PB: Is mda-7/IL-24 a ‘magic bullet’ for cancer? Cancer Res. 65:10128–10138. 2005. View Article : Google Scholar : PubMed/NCBI
3	Whitaker EL, Filippov VA and Duerksen-Hughes PJ: Interleukin 24: Mechanisms and therapeutic potential of an anti-cancer gene. Cytokine Growth Factor Rev. 23:323–331. 2012. View Article : Google Scholar : PubMed/NCBI
4	Ramesh R, Mhashilkar AM, Tanaka F, Saito Y, Branch CD, Sieger K, et al: Melanoma differentiation-associated gene 7/interleukin (IL)-24 is a novel ligand that regulates angiogenesis via the IL-22 receptor. Cancer Res. 63:5105–5113. 2003.PubMed/NCBI
5	Saeki T, Mhashilkar A, Swanson X, Zou-Yang XH, Sieger K, Kawabe S, et al: Inhibition of human lung cancer growth following adenovirus-mediated mda-7 gene expression in vivo. Oncogene. 21:4558–4566. 2002. View Article : Google Scholar : PubMed/NCBI
6	Nishikawa T, Ramesh R, Munshi A, Chada S and Meyn RE: Adenovirus-mediated mda-7 (IL24) gene therapy suppresses angiogenesis and sensitizes NSCLC xenograft tumors to radiation. Mol Ther. 9:818–828. 2004. View Article : Google Scholar : PubMed/NCBI
7	Caudell EG, Mumm JB, Poindexter N, Ekmekcioglu S, Mhashilkar AM, Yang XH, et al: The protein product of the tumor suppressor gene, melanoma differentiation-associated gene 7, exhibits immunostimulatory activity and is designated IL-24. J Immunol. 168:6041–6046. 2002. View Article : Google Scholar : PubMed/NCBI
8	Ramesh R, Ito I, Gopalan B, Saito Y, Mhashilkar AM and Chada S: Ectopic production of MDA-7/IL-24 inhibits invasion and migration of human lung cancer cells. Mol Ther. 9:510–518. 2004. View Article : Google Scholar : PubMed/NCBI
9	Sauane M, Su ZZ, Gupta P, Lebedeva IV, Dent P, Sarkar D and Fisher PB: Autocrine regulation of mda-7/IL-24 mediates cancer-specific apoptosis. Proc Natl Acad Sci USA. 105:9763–9768. 2008. View Article : Google Scholar : PubMed/NCBI
10	Hamed HA, Das SK, Sokhi UK, Park MA, Cruickshanks N, Archer K, et al: Combining histone deacetylase inhibitors with MDA-7/IL-24 enhances killing of renal carcinoma cells. Cancer Biol Ther. 14:1039–1049. 2013. View Article : Google Scholar : PubMed/NCBI
11	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
12	Tinoco G, Warsch S, Glück S, Avancha K and Montero AJ: Treating breast cancer in the 21st century: Emerging biological therapies. J Cancer. 4:117–132. 2013. View Article : Google Scholar : PubMed/NCBI
13	Patani N, Douglas-Jones A, Mansel R, Jiang W and Mokbel K: Tumour suppressor function of MDA-7/IL-24 in human breast cancer. Cancer Cell Int. 10:292010.PubMed/NCBI
14	Frewer NC, Ye L, Sun PH, Owen S, Ji K, Frewer KA, et al: Potential implication of IL-24 in lymphangiogenesis of human breast cancer. Int J Mol Med. 31:1097–1104. 2013.PubMed/NCBI
15	Sarkar D, Su ZZ, Vozhilla N, Park ES, Gupta P and Fisher PB: Dual cancer-specific targeting strategy cures primary and distant breast carcinomas in nude mice. Proc Natl Acad Sci USA. 102:14034–14039. 2005. View Article : Google Scholar : PubMed/NCBI
16	Sauane M, Gopalkrishnan RV, Choo HT, Gupta P, Lebedeva IV, Yacoub A, et al: Mechanistic aspects of mda-7/IL-24 cancer cell selectivity analysed via a bacterial fusion protein. Oncogene. 23:7679–7690. 2004. View Article : Google Scholar : PubMed/NCBI
17	Pei DS, Yang ZX, Zhang BF, Yin XX, Li LT, Li HZ and Zheng JN: Enhanced apoptosis-inducing function of MDA-7/IL-24 RGD mutant via the increased adhesion to tumor cells. J Interferon Cytokine Res. 32:66–73. 2012. View Article : Google Scholar : PubMed/NCBI
18	Weidner N: Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors. Breast Cancer Res Treat. 36:169–180. 1995. View Article : Google Scholar : PubMed/NCBI
19	Lebedeva IV, Emdad L, Su ZZ, Gupta P, Sauane M, Sarkar D, et al: mda-7/IL-24, novel anticancer cytokine: Focus on bystander antitumor, radiosensitization and antiangiogenic properties and overview of the phase I clinical experience (Review). Int J Oncol. 31:985–1007. 2007.PubMed/NCBI
20	Cunningham CC, Chada S, Merritt JA, Tong A, Senzer N, Zhang Y, et al: Clinical and local biological effects of an intratumoral injection of mda-7 (IL24; INGN 241) in patients with advanced carcinoma: a phase I study. Mol Ther. 11:149–159. 2005. View Article : Google Scholar
21	Xie Y, Sheng W, Xiang J, Ye Z, Zhu Y, Chen X and Yang J: Recombinant human IL-24 suppresses lung carcinoma cell growth via induction of cell apoptosis and inhibition of tumor angiogenesis. Cancer Biother Radiopharm. 23:310–320. 2008. View Article : Google Scholar : PubMed/NCBI
22	Yan S, Zhang H, Xie Y, Sheng W, Xiang J, Ye Z, et al: Recombinant human interleukin-24 suppresses gastric carcinoma cell growth in vitro and in vivo. Cancer Invest. 28:85–93. 2010. View Article : Google Scholar
23	Sauane M, Gupta P, Lebedeva IV, Su ZZ, Sarkar D, Randolph A, et al: N-glycosylation of MDA-7/IL-24 is dispensable for tumor cell-specific apoptosis and ‘bystander’ antitumor activity. Cancer Res. 66:11869–11877. 2006. View Article : Google Scholar : PubMed/NCBI
24	Danial NN and Korsmeyer SJ: Cell death: Critical control points. Cell. 116:205–219. 2004. View Article : Google Scholar : PubMed/NCBI
25	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
26	Welti J, Loges S, Dimmeler S and Carmeliet P: Recent molecular discoveries in angiogenesis and antiangiogenic therapies in cancer. J Clin Invest. 123:3190–3200. 2013. View Article : Google Scholar : PubMed/NCBI