Expression of the lysyl oxidase propeptide in hepatocellular carcinoma and its clinical relevance

Zheng,Ying; Wang,Xuemei; Wang,Haidong; Yan,Wei; Zhang,Quan; Chang,Xin

doi:10.3892/or.2014.3044

Expression of the lysyl oxidase propeptide in hepatocellular carcinoma and its clinical relevance

Authors:
- Ying Zheng
- Xuemei Wang
- Haidong Wang
- Wei Yan
- Quan Zhang
- Xin Chang
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Affiliations: Department of Anesthesia, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China, Department of Ultrasound, Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011, P.R. China, Department of Obstetrics and Gynecology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China, Department of Gastroenterology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China, Department of Oncology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China, Department of Imaging Medicine Center, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China
Published online on: February 20, 2014 https://doi.org/10.3892/or.2014.3044
Pages: 1669-1676

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Abstract

Lysyl oxidase is an important extracellular matrix remodeling enzyme and plays critical roles in tumor progression and development. Its tumor-suppressor activity has been shown to depend on the propeptide region. Previous studies have reported that the expression levels of lysyl oxidase propeptide (LOX-PP) are associated with cancer of the breast, pancreas, lung, prostate and gastrointestinal system. However, to date, the exact effects and molecular mechanisms of LOX-PP in hepatocellular carcinoma progression are still unclear. The present study aimed to investigate the expression and clinical significance of LOX-PP in human hepatocellular carcinoma. First, 42 cases of hepatocellular carcinoma and corresponding adjacent non-cancerous tissues (ANCTs) were collected, and the expression of LOX-PP in these samples was assessed by immunohistochemistry (IHC). The clinicopathological characteristics of all patients were recorded. Next, in in vitro studies, recombinant adenovirus LOX (ad-LOX-PP) was used to infect hepatocellular carcinoma cell lines to determine the function of LOX-PP. To determine whether ad-LOX-PP affects hepatocellular carcinoma cell survival, cell viability was examined by CCK-8 assay, and cell cycle progression was assessed by flow cytometry. We also investigated the effects of LOX-PP on the expression of cell cycle regulators (cyclin D1 and cyclin E) by western blot analysis. The migration and invasion capacities of hepatocellular carcinoma cells were observed by wound-healing and tranwell invasion assays. To further investigate how LOX-PP affects migration levels of matrix metallopeptidase (MMP)-2 and MMP-9 were assessed by western blot analysis. Additionally, markers of the PI3K and MAPK signaling pathway were detected to further confirm the mechanisms of LOX-PP. As a result, reduced expression of LOX-PP was found in hepatocellular carcinoma tissues, when compared with that in the ANCTs (15 vs. 83%, P<0.01), and its expression was associated with tumor stage and distant metastasis (each P<0.05). Proliferation in hepatocellular carcinoma cells was significantly decreased in the ad-LOX-PP group as indicated by CCK-8 assay. LOX-PP significantly reduced the expression of Ki-67, while prominent increases in the rate of apoptosis and cell cycle arrest were observed. Similarly, cell migration was significantly inhibited in the ad-LOX-PP group as evidenced by transwell invasion and wound-healing assays. The expression levels of MMP-2 and MMP-9 were attenuated in the ad-LOX-PP group, suggesting that LOX-PP inhibits hepatocellular carcinoma cell migration via down-regulation of MMPs expression. When LOX-PP expression was potentiated by an adenovirus containing LOX-PP, the expression of p-ERK was significantly downregulated, indicating that LOX-PP inhibits hepatocellular carcinoma cell proliferation and induces its apoptosis probably through downregulation of the MAPK/ERK pathway.

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1	El-Serag HB and Rudolph KL: Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology. 132:2557–2576. 2007. View Article : Google Scholar : PubMed/NCBI
2	Hsu HC, Huang AM, Lai PL, et al: Genetic alterations at the splice junction of p53 gene in human hepatocellular carcinoma. Hepatology. 19:122–128. 1994. View Article : Google Scholar : PubMed/NCBI
3	Hsu HC, Jeng YM, Mao TL, et al: β-catenin mutations are associated with a subset of low-stage hepatocellular carcinoma negative for hepatitis B virus and with favorable prognosis. Am J Pathol. 157:763–770. 2000.
4	Yuan JM, Govindarajan S, Ross RK, et al: Chronic infection with hepatitis G virus in relation to hepatocellular carcinoma among non-Asians in Los Angeles County, California. Cancer. 86:936–943. 1999. View Article : Google Scholar : PubMed/NCBI
5	Kagan HM and Trackman PC: Properties and function of lysyl oxidase. Am J Respir Cell Mol Biol. 5:206–210. 1991. View Article : Google Scholar : PubMed/NCBI
6	Kim YM, Kim EC and Kim Y: The human lysyl oxidase-like 2 protein functions as an amine oxidase toward collagen and elastin. Mol Biol Rep. 38:145–149. 2011. View Article : Google Scholar : PubMed/NCBI
7	Bais MV, Nugent MA, Stephens DN, et al: Recombinant lysyl oxidase propeptide protein inhibits growth and promotes apoptosis of pre-existing murine breast cancer xenografts. PLoS One. 7:e311882012. View Article : Google Scholar : PubMed/NCBI
8	Palamakumbura AH, Vora SR, Nugent MA, et al: Lysyl oxidase propeptide inhibits prostate cancer cell growth by mechanisms that target FGF-2-cell binding and signaling. Oncogene. 28:3390–3400. 2009. View Article : Google Scholar : PubMed/NCBI
9	Hurtado PA, Vora S, Sume SS, et al: Lysyl oxidase propeptide inhibits smooth muscle cell signaling and proliferation. Biochem Biophys Res Commun. 366:156–161. 2008. View Article : Google Scholar : PubMed/NCBI
10	Min C, Kirsch KH, Zhao Y, et al: The tumor suppressor activity of the lysyl oxidase propeptide reverses the invasive phenotype of Her-2/neu-driven breast cancer. Cancer Res. 67:1105–1112. 2007. View Article : Google Scholar : PubMed/NCBI
11	Palamakumbura AH, Jeay S, Guo Y, et al: The propeptide domain of lysyl oxidase induces phenotypic reversion of ras-transformed cells. J Biol Chem. 279:40593–40600. 2004. View Article : Google Scholar : PubMed/NCBI
12	Min C, Zhao Y, Romagnoli M, et al: Lysyl oxidase propeptide sensitizes pancreatic and breast cancer cells to doxorubicin-induced apoptosis. J Cell Biochem. 111:1160–1168. 2010. View Article : Google Scholar : PubMed/NCBI
13	Barker HE, Cox TR and Erler JT: The rationale for targeting the LOX family in cancer. Nat Rev Cancer. 12:540–552. 2012. View Article : Google Scholar : PubMed/NCBI
14	Edmondson HA and Steiner PE: Primary carcinoma of the liver: a study of 100 cases among 48,900 necropsies. Cancer. 7:462–503. 1954. View Article : Google Scholar : PubMed/NCBI
15	Yang YH, Chen K, Li B, et al: Estradiol inhibits osteoblast apoptosis via promotion of autophagy through the ER-ERK-mTOR pathway. Apoptosis. 18:1363–1375. 2013. View Article : Google Scholar : PubMed/NCBI
16	Chiu CC, Lin CH and Fang K: Etoposide (VP-16) sensitizes p53-deficient human non-small cell lung cancer cells to caspase-7-mediated apoptosis. Apoptosis. 10:643–650. 2005. View Article : Google Scholar : PubMed/NCBI
17	Novello C, Pazzaglia L, Cingolani C, et al: miRNA expression profile in human osteosarcoma: Role of miR-1 and miR-133b in proliferation and cell cycle control. Int J Oncol. 42:667–675. 2013.PubMed/NCBI
18	Aigelsreiter A, Ress AL, Bettermann K, et al: Low expression of the putative tumour suppressor spinophilin is associated with higher proliferative activity and poor prognosis in patients with hepatocellular carcinoma. Br J Cancer. 108:1830–1837. 2013. View Article : Google Scholar : PubMed/NCBI
19	Li B, Yang Y, Jiang S, et al: Adenovirus-mediated overexpression of BMP-9 inhibits human osteosarcoma cell growth and migration through downregulation of the PI3K/AKT pathway. Int J Oncol. 41:1809–1819. 2012.PubMed/NCBI
20	Liu ZL, Wang G, Peng AF, et al: Fatty acid synthase expression in osteosarcoma and its correlation with pulmonary metastasis. Oncol Lett. 4:878–882. 2012.PubMed/NCBI
21	Thompson L, Wang S, Tawfik O, et al: Effect of 25-hydroxyvitamin D₃ and 1 α,25 dihydroxyvitamin D₃ on differentiation and apoptosis of human osteosarcoma cell lines. J Orthop Res. 30:831–844. 2012.
22	Kamei S, Sakayama K, Tamashiro S, et al: Ketoprofen in topical formulation decreases the matrix metalloproteinase-2 expression and pulmonary metastatic incidence in nude mice with osteosarcoma. J Orthop Res. 27:909–915. 2009. View Article : Google Scholar : PubMed/NCBI
23	Korpi JT, Hagstrom J, Lehtonen N, et al: Expression of matrix metalloproteinases-2, −8, −13, −26, and tissue inhibitors of metalloproteinase-1 in human osteosarcoma. Surg Oncol. 20:e18–22. 2011.
24	Zhang Y, Song L, Cai L, et al: Effects of baicalein on apoptosis, cell cycle arrest, migration and invasion of osteosarcoma cells. Food Chem Toxicol. 53C:325–333. 2012.PubMed/NCBI
25	Gu X, Xing L, Shi G, et al: The circadian mutation PER2^S662G is linked to cell cycle progression and tumorigenesis. Cell Death Differ. 19:397–405. 2012.
26	Oka K, Ohya-Shimada W, Mizuno S, et al: Up-regulation of cyclin-E₁ via proline-mTOR pathway is responsible for HGF-mediated G₁/S progression in the primary culture of rat hepatocytes. Biochem Biophys Res Commun. 435:120–125. 2013.PubMed/NCBI
27	Rasmussen N and Rathmell WK: Looking beyond inhibition of VEGF/mTOR: emerging targets for renal cell carcinoma drug development. Curr Clin Pharmacol. 6:199–206. 2011. View Article : Google Scholar : PubMed/NCBI
28	Van Waes C: Nuclear factor-κB in development, prevention, and therapy of cancer. Clin Cancer Res. 13:1076–1082. 2007.
29	Peng L, Liu A, Shen Y, et al: Antitumor and anti-angiogenesis effects of thymoquinone on osteosarcoma through the NF-κB pathway. Oncol Rep. 29:571–578. 2013.PubMed/NCBI
30	Jeng YM, Chang CC, Hu FC, et al: RNA-binding protein insulin-like growth factor II mRNA-binding protein 3 expression promotes tumor invasion and predicts early recurrence and poor prognosis in hepatocellular carcinoma. Hepatology. 48:1118–1127. 2008. View Article : Google Scholar
31	Chen YL, Wang TH, Hsu HC, et al: Overexpression of CTHRC1 in hepatocellular carcinoma promotes tumor invasion and predicts poor prognosis. PLoS One. 8:e703242013. View Article : Google Scholar : PubMed/NCBI
32	Sanchez-Morgan N, Kirsch KH, Trackman PC, et al: The lysyl oxidase propeptide interacts with the receptor-type protein tyrosine phosphatase kappa and inhibits β-catenin transcriptional activity in lung cancer cells. Mol Cell Biol. 31:3286–3297. 2011.PubMed/NCBI
33	Yoon JH, Park JK, Kang YH, et al: Lysyl oxidase G473A polymorphism is closely associated with susceptibility to gastric cancer in a South Korean population. APMIS. 119:762–768. 2011. View Article : Google Scholar : PubMed/NCBI
34	Yu Z, Sato S, Trackman PC, et al: Blimp1 activation by AP-1 in human lung cancer cells promotes a migratory phenotype and is inhibited by the lysyl oxidase propeptide. PLoS One. 7:e332872012. View Article : Google Scholar : PubMed/NCBI
35	Barcellos-Hoff MH and Ravani SA: Irradiated mammary gland stroma promotes the expression of tumorigenic potential by unirradiated epithelial cells. Cancer Res. 60:1254–1260. 2000.PubMed/NCBI
36	Wiseman BS and Werb Z: Stromal effects on mammary gland development and breast cancer. Science. 296:1046–1049. 2002. View Article : Google Scholar : PubMed/NCBI