Secreted recombinant human IL-24 protein inhibits the proliferation of esophageal squamous cell carcinoma Eca-109 cells in vitro and in vivo

Ma,Qunfeng; Jin,Bangming; Zhang,Yao; Shi,Yinan; Zhang,Chi; Luo,Dan; Wang,Pengkun; Duan,Cuimi; Song,Heyu; Li,Xue; Deng,Xuefeng; Chen,Zhinan; Wang,Ziling; Jiang,Hong; Liu,Yan

doi:10.3892/or.2016.4633

Secreted recombinant human IL-24 protein inhibits the proliferation of esophageal squamous cell carcinoma Eca-109 cells in vitro and in vivo

Authors:
- Qunfeng Ma
- Bangming Jin
- Yao Zhang
- Yinan Shi
- Chi Zhang
- Dan Luo
- Pengkun Wang
- Cuimi Duan
- Heyu Song
- Xue Li
- Xuefeng Deng
- Zhinan Chen
- Ziling Wang
- Hong Jiang
- Yan Liu
View Affiliations

Affiliations: Department of Thoracic Surgery, Affiliated Hospital of the Academy of Military Medical Sciences, Fengtai, Beijing 100071, P.R. China, College of Life Science and Bioengineering, School of Science, Beijing Jiaotong University, Haidian, Beijing 100044, P.R. China, Department of Biological Diagnosis, Beijing Institute of Basic Medical Sciences, Haidian, Beijing 100850, P.R. China, Cell Engineering Research Center, The Fourth Military Medical University, Xicheng, Xi'an, Shaanxi 710032, P.R. China, College of Life Science, Southwest University, Beibei, Chongqing 400715, P.R. China
Published online on: February 24, 2016 https://doi.org/10.3892/or.2016.4633
Pages: 2681-2690

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

Interleukin-24 (IL-24) displays cancer-specific apoptosis-inducing properties in a broad spectrum of human tumors without harmful effects on normal cells. The human IL-24 protein is secreted as a glycosylated protein and functions as a pro-Th1 cytokine and a potent antiangiogenic molecule. However, the function of secreted recombinant human IL-24 (srhIL-24) protein in esophageal squamous cell carcinoma (ESCC) cells has not been studied. In the present study, we prepared a stable site-specific-integrated cell line, Flp-InTMCHO/IL-24 (FCHO/IL-24), which secreted rhIL-24 at a higher level than three random-integrated cell lines. In vitro, we identified that the purified srhIL-24 inhibited proliferation and induced the apoptosis of ESCC Eca-109 cells and activated STAT3, which was related with the IL-20 receptors. In vivo, the tumorigenicity of Eca-109 cells was significantly inhibited by s.c. injection of FCHO/IL-24 cells. Decreased tumor microvessel density and an increased number of TUNEL-positive tumor cells were associated with tumor growth inhibition, indicating the presence of antiangiogenic activity and induction of apoptotic activity. In summary, the present study demonstrated that srhIL-24 induced growth inhibition and apoptosis in ESCC Eca-109 cells in vitro and in vivo, which may be mediated by the receptor pathway.

View Figures

View References

1	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
2	Wang JB, Fan JH, Liang H, Li J, Xiao HJ, Wei WQ, Dawsey SM, Qiao YL and Boffetta P: Attributable causes of esophageal cancer incidence and mortality in China. PLoS One. 7:e422812012. View Article : Google Scholar : PubMed/NCBI
3	Burrows WM: Gastrointestinal function and related problems following esophagectomy. Semin Thorac Cardiovasc Surg. 16:142–151. 2004. View Article : Google Scholar : PubMed/NCBI
4	Xu YW, Peng YH, Chen B, Wu ZY, Wu JY, Shen JH, Zheng CP, Wang SH, Guo HP, Li EM, et al: Autoantibodies as potential biomarkers for the early detection of esophageal squamous cell carcinoma. Am J Gastroenterol. 109:36–45. 2014. View Article : Google Scholar :
5	Jiang H, Lin JJ, Su ZZ, Goldstein NI and Fisher PB: Subtraction hybridization identifies a novel melanoma differentiation associated gene, mda-7, modulated during human melanoma differentiation, growth and progression. Oncogene. 11:2477–2486. 1995.PubMed/NCBI
6	Gupta P, Su ZZ, Lebedeva IV, Sarkar D, Sauane M, Emdad L, Bachelor MA, Grant S, Curiel DT, Dent P, et al: mda-7/IL-24: Multifunctional cancer-specific apoptosis-inducing cytokine. Pharmacol Ther. 111:596–628. 2006. View Article : Google Scholar : PubMed/NCBI
7	Caudell EG, Mumm JB, Poindexter N, Ekmekcioglu S, Mhashilkar AM, Yang XH, Retter MW, Hill P, Chada S and Grimm EA: 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	Fisher PB: Is mda-7/IL-24 a ʻmagic bulletʼ for cancer? Cancer Res. 65:10128–10138. 2005. View Article : Google Scholar : PubMed/NCBI
9	Su ZZ, Lebedeva IV, Sarkar D, Gopalkrishnan RV, Sauane M, Sigmon C, Yacoub A, Valerie K, Dent P and Fisher PB: Melanoma differentiation associated gene-7, mda-7/IL-24, selectively induces growth suppression, apoptosis and radiosensitization in malignant gliomas in a p53-independent manner. Oncogene. 22:1164–1180. 2003. View Article : Google Scholar : PubMed/NCBI
10	Wang J, Fan X, Bao L and Du L: Construction of eukaryotic expression vector of EGFRi-IL-24 recombinant gene. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 27:395–399. 2010.In Chinese. PubMed/NCBI
11	Sieger KA, Mhashilkar AM, Stewart A, Sutton RB, Strube RW, Chen SY, Pataer A, Swisher SG, Grimm EA, Ramesh R, et al: The tumor suppressor activity of MDA-7/IL-24 is mediated by intracellular protein expression in NSCLC cells. Mol Ther. 9:355–367. 2004. View Article : Google Scholar : PubMed/NCBI
12	Ekmekcioglu S, Ellerhorst JA, Mumm JB, Zheng M, Broemeling L, Prieto VG, Stewart AL, Mhashilkar AM, Chada S and Grimm EA: Negative association of melanoma differentiation-associated gene (mda-7) and inducible nitric oxide synthase (iNOS) in human melanoma: MDA-7 regulates iNOS expression in melanoma cells. Mol Cancer Ther. 2:9–17. 2003. View Article : Google Scholar : PubMed/NCBI
13	Zheng M, Bocangel D, Ramesh R, Ekmekcioglu S, Poindexter N, Grimm EA and Chada S: Interleukin-24 overcomes temozolo-mide resistance and enhances cell death by down-regulation of O⁶-methylguanine-DNA methyltransferase in human melanoma cells. Mol Cancer Ther. 7:3842–3851. 2008. View Article : Google Scholar : PubMed/NCBI
14	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
15	Chada S, Bocangel D, Ramesh R, Grimm EA, Mumm JB, Mhashilkar AM and Zheng M: mda-7/IL24 kills pancreatic cancer cells by inhibition of the Wnt/PI3K signaling pathways: Identification of IL-20 receptor-mediated bystander activity against pancreatic cancer. Mol Ther. 11:724–733. 2005. View Article : Google Scholar : PubMed/NCBI
16	Ramesh R, Mhashilkar AM, Tanaka F, Saito Y, Branch CD, Sieger K, Mumm JB, Stewart AL, Boquoi A, Dumoutier L, 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
17	Zheng M, Bocangel D, Doneske B, Mhashilkar A, Ramesh R, Hunt KK, Ekmekcioglu S, Sutton RB, Poindexter N, Grimm EA, et al: Human interleukin 24 (MDA-7/IL-24) protein kills breast cancer cells via the IL-20 receptor and is antagonized by IL-10. Cancer Immunol Immunother. 56:205–215. 2007. View Article : Google Scholar
18	Lebedeva IV, Su ZZ, Sarkar D and Fisher PB: Restoring apoptosis as a strategy for cancer gene therapy: Focus on p53 and mda-7. Semin Cancer Biol. 13:169–178. 2003. View Article : Google Scholar : PubMed/NCBI
19	Su Z, Emdad L, Sauane M, Lebedeva IV, Sarkar D, Gupta P, James CD, Randolph A, Valerie K, Walter MR, et al: Unique aspects of mda-7/IL-24 antitumor bystander activity: Establishing a role for secretion of MDA-7/IL-24 protein by normal cells. Oncogene. 24:7552–7566. 2005. View Article : Google Scholar : PubMed/NCBI
20	Ma G, Kawamura K, Shan Y, Okamoto S, Li Q, Namba M, Shingyoji M, Tada Y, Tatsumi K, Hiroshima K, et al: Combination of adenoviruses expressing melanoma differentiation-associated gene-7 and chemotherapeutic agents produces enhanced cyto-toxicity on esophageal carcinoma. Cancer Gene Ther. 21:31–37. 2014. View Article : Google Scholar : PubMed/NCBI
21	Ma Q, Deng X, Jin B, Zhang Y, Luo D, Song H, Wang P, Zhang C, Li X, Shi Y, et al: A novel human interleukin-24 peptide created by computer-guided design contributes to suppression of proliferation in esophageal squamous cell carcinoma Eca-109 cells. Oncol Rep. 33:193–200. 2015.
22	Yang J, Zhang W, Liu K, Jing S, Guo G, Luo P and Zou Q: Expression, purification, and characterization of recombinant human interleukin 24 in Escherichia coli. Protein Expr Purif. 53:339–345. 2007. View Article : Google Scholar : PubMed/NCBI
23	Chada S, Mhashilkar AM, Ramesh R, Mumm JB, Sutton RB, Bocangel D, Zheng M, Grimm EA and Ekmekcioglu S: Bystander activity of Ad-mda7: Human MDA-7 protein kills melanoma cells via an IL-20 receptor-dependent but STAT3-independent mechanism. Mol Ther. 10:1085–1095. 2004. View Article : Google Scholar : PubMed/NCBI
24	Aggarwal S, Takada Y, Mhashilkar AM, Sieger K, Chada S and Aggarwal BB: Melanoma differentiation-associated gene-7/IL-24 gene enhances NF-kappa B activation and suppresses apoptosis induced by TNF. J Immunol. 173:4368–4376. 2004. View Article : Google Scholar : PubMed/NCBI
25	Yacoub A, Mitchell C, Hong Y, Gopalkrishnan RV, Su ZZ, Gupta P, Sauane M, Lebedeva IV, Curiel DT, Mahasreshti PJ, et al: MDA-7 regulates cell growth and radiosensitivity in vitro of primary (non-established) human glioma cells. Cancer Biol Ther. 3:739–751. 2004. View Article : Google Scholar : PubMed/NCBI
26	Kreis S, Philippidou D, Margue C, Rolvering C, Haan C, Dumoutier L, Renauld JC and Behrmann I: Recombinant interleukin-24 lacks apoptosis-inducing properties in melanoma cells. PLoS One. 2:e13002007. View Article : Google Scholar : PubMed/NCBI
27	Mumm JB, Ekmekcioglu S, Poindexter NJ, Chada S and Grimm EA: Soluble human MDA-7/IL-24: Characterization of the molecular form(s) inhibiting tumor growth and stimulating monocytes. J Interferon Cytokine Res. 26:877–886. 2006. View Article : Google Scholar
28	Dash R, Bhutia SK, Azab B, Su ZZ, Quinn BA, Kegelmen TP, Das SK, Kim K, Lee SG, Park MA, et al: mda-7/IL-24: A unique member of the IL-10 gene family promoting cancer-targeted toxicity. Cytokine Growth Factor Rev. 21:381–391. 2010. View Article : Google Scholar : PubMed/NCBI
29	Saeki T, Mhashilkar A, Swanson X, Zou-Yang XH, Sieger K, Kawabe S, Branch CD, Zumstein L, Meyn RE, Roth JA, 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
30	Tamai H, Miyake K, Yamaguchi H, Takatori M, Dan K, Inokuchi K and Shimada T: AAV8 vector expressing IL24 efficiently suppresses tumor growth mediated by specific mechanisms in MLL/AF4-positive ALL model mice. Blood. 119:64–71. 2012. View Article : Google Scholar
31	Gu J, Chen X, Xin H, Fang X and Sha X: Serum-resistant complex nanoparticles functionalized with imidazole-rich polypeptide for gene delivery to pulmonary metastatic melanoma. Int J Pharm. 461:559–569. 2014. View Article : Google Scholar