Ubenimex inhibits cell proliferation, migration and invasion by inhibiting the expression of APN and inducing autophagic cell death in prostate cancer cells

Wang,Xiaoqing; Niu,Zhihong; Jia,Yang; Cui,Meng; Han,Liping; Zhang,Yongfei; Liu,Zheng; Bi,Dongbin; Liu,Shuai

doi:10.3892/or.2016.4611

Ubenimex inhibits cell proliferation, migration and invasion by inhibiting the expression of APN and inducing autophagic cell death in prostate cancer cells

Authors:
- Xiaoqing Wang
- Zhihong Niu
- Yang Jia
- Meng Cui
- Liping Han
- Yongfei Zhang
- Zheng Liu
- Dongbin Bi
- Shuai Liu
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Affiliations: Department of Urology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China, Shandong University, Jinan, Shandong 250000, P.R. China, Department of Neurology, Shandong Police Hospital, Jinan, Shandong 250000, P.R. China, Department of Dermatology, Mount Qianfu Attached Hospital of Shandong University, Jinan, Shandong 250000, P.R. China
Published online on: February 3, 2016 https://doi.org/10.3892/or.2016.4611
Pages: 2121-2130

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Abstract

Prostate cancer is the second most frequently diagnosed cancer in males worldwide and is commonly associated with metastasis. Moreover, in prostate cancer, aminopeptidase N (APN) expression is closely correlated with metastasis. Ubenimex, an APN inhibitor, is widely used as an adjunct therapy for cancer, enhancing the function of immunocompetent cells and conferring antitumor effects. However, due to the low expression of APN, it is rarely used to treat prostate cancer. Recently, the induction of autophagy as a molecular mechanism has been strongly connected with tumor cell death. Thus, we investigated whether ubenimex could inhibit cell proliferation, migration and invasion by downregulating APN expression to induce autophagic cell death in prostate cancer cells. The LNCaP and PC-3 cell lines were treated with different doses of ubenimex. Cell viability was measured using growth curve analysis and WST-8 proliferation assay. Autophagic cell death was assessed using fluorescence microscopy and acridine orange/ethidium bromide (AO/EB) staining. Protein expression was assessed by immunofluorescence and western blot analyses. Autophagosomes were evaluated using transmission electron microscopy. Wound-healing migration assays were performed to determine the migratory ability of the PC-3 cells. In addition, nude mice were used in the present study to examine PC-3 cell proliferation in vivo. The results revealed that APN expression differed between the metastatic and non-metastatic prostate cancer cells. In addition, ubenimex inhibited APN expression in the prostate cancer cells. Ubenimex increased prostate cancer cell death, as determined using the lactate dehydrogenase (LDH) cytotoxicity assay. This effect was accompanied by increased levels of LC3B. Furthermore, ubenimex inhibited PC-3 cell proliferation in vivo and in vitro. Ubenimex inhibited the cell migration and invasion in prostate cancer cells by downregulating APN expression. Finally, ubenimex induced autophagic cell death in both metastatic and non-metastatic prostate cancer cells. Based on these results, ubenimex appears to be an excellent adjunctive therapy for the treatment of prostate cancer.

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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	Damber JE and Aus G: Prostate cancer. Lancet. 371:1710–1721. 2008. View Article : Google Scholar : PubMed/NCBI
3	Liu S, Xie F, Wang H, Liu Z, Liu X, Sun L and Niu Z: Ubenimex inhibits cell proliferation, migration and invasion in renal cell carcinoma: The effect is autophagy-associated. Oncol Rep. 33:1372–1380. 2015.PubMed/NCBI
4	Ikeda N, Nakajima Y, Tokuhara T, Hattori N, Sho M, Kanehiro H and Miyake M: Clinical significance of aminopeptidase N/CD13 expression in human pancreatic carcinoma. Clin Cancer Res. 9:1503–1508. 2003.PubMed/NCBI
5	Tokuhara T, Hattori N, Ishida H, Hirai T, Higashiyama M, Kodama K and Miyake M: Clinical significance of aminopeptidase N in non-small cell lung cancer. Clin Cancer Res. 12:3971–3978. 2006. View Article : Google Scholar : PubMed/NCBI
6	Bogenrieder T, Finstad CL, Freeman RH, Papandreou CN, Scher HI, Albino AP, Reuter VE and Nanus DM: Expression and localization of aminopeptidase A, aminopeptidase N, and dipeptidyl peptidase IV in benign and malignant human prostate tissue. Prostate. 33:225–232. 1997. View Article : Google Scholar : PubMed/NCBI
7	Liu AY, Roudier MP and True LD: Heterogeneity in primary and metastatic prostate cancer as defined by cell surface CD profile. Am J Pathol1. 165:1543–1556. 2004. View Article : Google Scholar
8	Dall'Era MA, True LD, Siegel AF, Porter MP, Sherertz TM and Liu AY: Differential expression of CD10 in prostate cancer and its clinical implication. BMC Urol. 7:32007. View Article : Google Scholar : PubMed/NCBI
9	Ishii K, Usui S, Sugimura Y, Yoshida S, Hioki T, Tatematsu M, Yamamoto H and Hirano K: Aminopeptidase N regulated by zinc in human prostate participates in tumor cell invasion. Int J Cancer. 92:49–54. 2001. View Article : Google Scholar : PubMed/NCBI
10	Fontijn D, Duyndam MC, van Berkel MP, Yuana Y, Shapiro LH, Pinedo HM, Broxterman HJ and Boven E: CD13/Aminopeptidase N overexpression by basic fibroblast growth factor mediates enhanced invasiveness of 1F6 human melanoma cells. Br J Cancer. 94:1627–1636. 2006.PubMed/NCBI
11	Mina-Osorio P: The moonlighting enzyme CD13: Old and new functions to target. Trends Mol Med. 14:361–371. 2008. View Article : Google Scholar : PubMed/NCBI
12	Horita H, Frankel AE and Thorburn A: Acute myeloid leukemia-targeted toxin activates both apoptotic and necroptotic death mechanisms. PLoS One. 3:e39092008. View Article : Google Scholar : PubMed/NCBI
13	Mujtaba SF, Dwivedi A, Yadav N, Ch R, Kushwaha HN, Mudiam MK, Singh G and Ray RS: Superoxide mediated photo-modification and DNA damage induced apoptosis by Benz(a) anthracene via mitochondrial mediated pathway. J Photochem Photobiol B. 142:92–102. 2015. View Article : Google Scholar
14	Sørensen KD, Abildgaard MO, Haldrup C, Ulhøi BP, Kristensen H, Strand S, Parker C, Høyer S, Borre M and Ørntoft TF: Prognostic significance of aberrantly silenced ANPEP expression in prostate cancer. Br J Cancer. 108:420–428. 2013. View Article : Google Scholar : PubMed/NCBI
15	Ishii K, Usui S, Sugimura Y, Yamamoto H, Yoshikawa K and Hirano K: Inhibition of aminopeptidase N (AP-N) and urokinase-type plasminogen activator (uPA) by zinc suppresses the invasion activity in human urological cancer cells. Biol Pharm Bull. 24:226–230. 2001. View Article : Google Scholar : PubMed/NCBI
16	Hashida H, Takabayashi A, Kanai M, Adachi M, Kondo K, Kohno N, Yamaoka Y and Miyake M: Aminopeptidase N is involved in cell motility and angiogenesis: Its clinical significance in human colon cancer. Gastroenterology. 122:376–386. 2002. View Article : Google Scholar : PubMed/NCBI
17	Murakami H, Yokoyama A, Kondo K, Nakanishi S, Kohno N and Miyake M: Circulating aminopeptidase N/CD13 is an independent prognostic factor in patients with non-small cell lung cancer. Clin Cancer Res. 11:8674–8679. 2005. View Article : Google Scholar : PubMed/NCBI
18	Wickström M, Larsson R, Nygren P, Gullbo J and Aminopeptidase N: Aminopeptidase N (CD13) as a target for cancer chemotherapy. Cancer Sci. 102:501–508. 2011. View Article : Google Scholar : PubMed/NCBI
19	Ichinose Y, Genka K, Koike T, Kato H, Watanabe Y, Mori T, Iioka S, Sakuma A and Ohta M; NK421 Lung Cancer Surgery Group: Randomized double-blind placebo-controlled trial of bestatin in patients with resected stage I squamous-cell lung carcinoma. J Natl Cancer Inst. 95:605–610. 2003. View Article : Google Scholar : PubMed/NCBI
20	Wakita A, Ohtake S, Takada S, Yagasaki F, Komatsu H, Miyazaki Y, Kubo K, Kimura Y, Takeshita A, Adachi Y, et al: Randomized comparison of fixed-schedule versus response-oriented individualized induction therapy and use of ubenimex during and after consolidation therapy for elderly patients with acute myeloid leukemia: The JALSG GML200 Study. Int J Hematol. 96:84–93. 2012. View Article : Google Scholar : PubMed/NCBI
21	Tsukamoto H, Shibata K, Kajiyama H, Terauchi M, Nawa A, Kikkawa F and Aminopeptidase N: Aminopeptidase N (APN)/CD13 inhibitor, Ubenimex, enhances radiation sensitivity in human cervical cancer. BMC Cancer. 8:742008. View Article : Google Scholar : PubMed/NCBI
22	Xu JW, Li CG, Huang XE, Li Y and Huo JG: Ubenimex capsule improves general performance and chemotherapy related toxicity in advanced gastric cancer cases. Asian Pac J Cancer Prev. 12:985–987. 2011.PubMed/NCBI
23	Yang ZJ, Chee CE, Huang S and Sinicrope F: Autophagy modulation for cancer therapy. Cancer Biol Ther. 11:169–176. 2011. View Article : Google Scholar : PubMed/NCBI
24	Eum KH and Lee M: Crosstalk between autophagy and apoptosis in the regulation of paclitaxel-induced cell death in v-Ha-ras- transformed fibroblasts. Mol Cell Biochem. 348:61–68. 2011. View Article : Google Scholar