Silencing of hypoxia‑inducible adrenomedullin using RNA interference attenuates hepatocellular carcinoma cell growth in vivo

Li,Fenbao; Yang,Ruimin; Zhang,Xizhong; Liu,Aiguang; Zhao,Yongli; Guo,Yingchang

doi:10.3892/mmr.2014.2320

Silencing of hypoxia‑inducible adrenomedullin using RNA interference attenuates hepatocellular carcinoma cell growth in vivo

Authors:
- Fenbao Li
- Ruimin Yang
- Xizhong Zhang
- Aiguang Liu
- Yongli Zhao
- Yingchang Guo
View Affiliations

Affiliations: Department of Interventional Therapy, First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, P.R. China
Published online on: June 13, 2014 https://doi.org/10.3892/mmr.2014.2320
Pages: 1295-1302

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

Adrenomedullin (ADM) is an angiogenic peptide that has been shown to increase the risk of endometrial hyperplasia and to promote tumor cell survival following hypoxia. ADM may induce microvessel proliferation and partially decrease hypoxia in solid tumors, thus contributing to the proliferation of tumor cells, as well as tumor invasion and metastasis. However, the impact of hypoxia‑induced ADM expression on hepatocellular carcinoma (HCC) cells requires further elucidation. In the present study it was found that the levels of ADM mRNA in tumor tissue from patients with HCC were significantly increased compared with the mRNA levels in adjacent non‑tumorous liver tissue. Under hypoxic conditions, the mRNA and protein levels of ADM, as well as those of the cancer‑promoting genes vascular endothelial growth factor and hypoxia‑inducible factor 1α, were significantly elevated in a time‑dependent manner in three human HCC cell lines. In addition, knockdown of ADM expression using short hairpin RNA (shRNA) in SMMC‑7721 cells resulted in apoptosis that was not observed in untransfected cells. Furthermore, combined treatment with cisplatin and ADM‑shRNA significantly decreased tumor growth in vivo compared with treatment with cisplatin or ADM‑shRNA alone. These data demonstrate that ADM acts as a critical promoter of cell cycle progression in HCC and that the inhibition of ADM may be an effective interventional therapeutic strategy in HCC.

View Figures

View References

1	Moradpour D and Blum HE: Pathogenesis of hepatocellular carcinoma. Eur J Gastroenterol Hepatol. 17:477–483. 2005. View Article : Google Scholar
2	Tanaka S and Arii S: Current status and perspective of antiangiogenic therapy for cancer: hepatocellular carcinoma. Int J Clin Oncol. 11:82–89. 2006. View Article : Google Scholar : PubMed/NCBI
3	Gwak GY, Yoon JH, Kim KM, Lee HS, Chung JW and Gores GJ: Hypoxia stimulates proliferation of human hepatoma cells through the induction of hexokinase II expression. J Hepatol. 42:358–364. 2005. View Article : Google Scholar : PubMed/NCBI
4	Liotta LA, Steeg PS and Stetler-Stevenson WG: Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell. 64:327–336. 1991. View Article : Google Scholar : PubMed/NCBI
5	Hanahan D and Folkman J: Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell. 86:353–364. 1996. View Article : Google Scholar : PubMed/NCBI
6	Carmeliet P and Jain RK: Angiogenesis in cancer and other diseases. Nature. 407:249–257. 2000. View Article : Google Scholar : PubMed/NCBI
7	Dvorak HF: Vascular permeability factor/vascular endothelial growth factor: a critical cytokine in tumor angiogenesis and a potential target for diagnosis and therapy. J Clin Oncol. 20:4368–4380. 2002. View Article : Google Scholar
8	Kitamura K, Kangawa K, Kawamoto M, et al: Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun. 192:553–560. 1993. View Article : Google Scholar : PubMed/NCBI
9	Nikitenko LL, Fox SB, Kehoe S, Rees MC and Bicknell R: Adrenomedullin and tumour angiogenesis. Br J Cancer. 94:1–7. 2006. View Article : Google Scholar
10	Jiménez N, Abasolo I, Jongsma J, et al: Androgen-independent expression of adrenomedullin and peptidylglycine alpha-amidating monooxygenase in human prostatic carcinoma. Mol Carcinog. 38:14–24. 2003.PubMed/NCBI
11	Li Z, Takeuchi S, Otani T and Maruo T: Implications of adrenomedullin expression in the invasion of squamous cell carcinoma of the uterine cervix. Int J Clin Oncol. 6:263–270. 2001. View Article : Google Scholar : PubMed/NCBI
12	Park SC, Yoon JH, Lee JH, et al: Hypoxia-inducible adrenomedullin accelerates hepatocellular carcinoma cell growth. Cancer Lett. 271:314–322. 2008. View Article : Google Scholar : PubMed/NCBI
13	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
14	Deville JL, Salas S, Figarella-Branger D, et al: Adrenomedullin as a therapeutic target in angiogenesis. Expert Opin Ther Targets. 14:1059–1072. 2010. View Article : Google Scholar : PubMed/NCBI
15	Wu SD, Ma YS, Fang Y, Liu LL, Fu D and Shen XZ: Role of the microenvironment in hepatocellular carcinoma development and progression. Cancer Treat Rev. 38:218–225. 2012. View Article : Google Scholar : PubMed/NCBI
16	Zudaire E, Martínez A and Cuttitta F: Adrenomedullin and cancer. Regul Pept. 112:175–183. 2003. View Article : Google Scholar
17	Oehler MK, Norbury C, Hague S, Rees MC and Bicknell R: Adrenomedullin inhibits hypoxic cell death by upregulation of Bcl-2 in endometrial cancer cells: a possible promotion mechanism for tumour growth. Oncogene. 20:2937–2945. 2001. View Article : Google Scholar : PubMed/NCBI
18	Oladipupo S, Hu S, Kovalski J, et al: VEGF is essential for hypoxia-inducible factor-mediated neovascularization but dispensable for endothelial sprouting. Proc Natl Acad Sci USA. 108:13264–13269. 2011. View Article : Google Scholar : PubMed/NCBI
19	Frede S, Freitag P, Otto T, Heilmaier C and Fandrey J: The proinflammatory cytokine interleukin 1beta and hypoxia cooperatively induce the expression of adrenomedullin in ovarian carcinoma cells through hypoxia inducible factor 1 activation. Cancer Res. 65:4690–4697. 2005. View Article : Google Scholar
20	Ribatti D, Nico B, Spinazzi R, Vacca A and Nussdorfer GG: The role of adrenomedullin in angiogenesis. Peptides. 26:1670–1675. 2005. View Article : Google Scholar : PubMed/NCBI
21	Jung KH, Zheng HM, Jeong Y, et al: Suppression of tumor proliferation and angiogenesis of hepatocellular carcinoma by HS-104, a novel phosphoinositide 3-kinase inhibitor. Cancer Lett. 328:176–187. 2013. View Article : Google Scholar : PubMed/NCBI
22	Harris AL: Hypoxia - a key regulatory factor in tumour growth. Nat Rev Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
23	Kim Y, Lin Q, Glazer PM and Yun Z: Hypoxic tumor microenvironment and cancer cell differentiation. Curr Mol Med. 9:425–434. 2009. View Article : Google Scholar : PubMed/NCBI
24	Tang N, Wang L, Esko J, et al: Loss of HIF-1alpha in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis. Cancer Cell. 6:485–495. 2004. View Article : Google Scholar : PubMed/NCBI
25	Carmeliet P, Dor Y, Herbert JM, et al: Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis. Nature. 394:485–490. 1998. View Article : Google Scholar : PubMed/NCBI
26	Iwase T, Nagaya N, Fujii T, et al: Adrenomedullin enhances angiogenic potency of bone marrow transplantation in a rat model of hindlimb ischemia. Circulation. 111:356–362. 2005. View Article : Google Scholar : PubMed/NCBI
27	Hazan RB, Phillips GR, Qiao RF, Norton L and Aaronson SA: Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis. J Cell Biol. 148:779–790. 2000. View Article : Google Scholar : PubMed/NCBI
28	Gwak GY, Yoon JH, Yu SJ, et al: Anti-apoptotic N-cadherin signaling and its prognostic implication in human hepatocellular carcinomas. Oncol Rep. 15:1117–1123. 2006.PubMed/NCBI
29	Shichiri M and Hirata Y: Regulation of cell growth and apoptosis by adrenomedullin. Hypertens Res. 26(Suppl): S9–S14. 2003. View Article : Google Scholar : PubMed/NCBI
30	Pang X, Shang H, Deng B, Wen F and Zhang Y: The interaction of adrenomedullin and macrophages induces ovarian cancer cell migration via activation of RhoA signaling pathway. Int J Mol Sci. 14:2774–2787. 2013. View Article : Google Scholar : PubMed/NCBI