Pim-3 promotes human pancreatic cancer growth by regulating tumor vasculogenesis

Liu,Bin; Wang,Zhen; Li,Hong-Yu; Zhang,Bo; Ping,Bo; Li,Ying-Yi

doi:10.3892/or.2014.3158

Pim-3 promotes human pancreatic cancer growth by regulating tumor vasculogenesis

Authors:
- Bin Liu
- Zhen Wang
- Hong-Yu Li
- Bo Zhang
- Bo Ping
- Ying-Yi Li
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Affiliations: Cancer Research Institute, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, P.R. China, Department of Gastroenterology, Shenyang General Hospital, Shenyang, Liaoning, P.R. China, Department of Pancreas and Hepatobiliary Surgery, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, P.R. China, Department of Pathology, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, P.R. China
Published online on: April 25, 2014 https://doi.org/10.3892/or.2014.3158
Pages: 2625-2634

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Abstract

Pim-3, a proto-oncogene with serine/threonine kinase activity, is aberrantly expressed in malignant lesions, but not in normal pancreatic tissues. To assess the role of Pim-3 in human pancreatic carcinogenesis in vivo and to determine the underlying Pim-3 signaling regulatory mechanisms, we established MiaPaca-2 cells overexpressing wild-type Pim-3 or Pim-3 kinase dead mutants (K69M-Pim-3) as well as PCI55 cells stably expressing Pim-3 shRNA or scrambled shRNA in a tetracycline-inducible manner. In addition, we conducted studies utilizing a nude mouse tumor xenograft model. Our results demonstrated that cells stably overexpressing wild-type Pim-3 exhibited functionally enhanced phosphorylation of Bad at Ser112 and increased proliferation. In contrast, the stable inactivation of Pim-3 by K69M-Pim-3 or silencing of Pim-3 expression by Pim-3 shRNA resulted in functionally decreased phosphorylation of Bad at Ser112 and higher apoptotic cells. Following subcutaneous injection of these stable cell lines, nude mice injected with Pim-3 overexpressing cells developed 100% subcutaneous tumors, together with increased PCNA-positive cells and enhanced intratumoral CD31-positive vascular areas. On the other hand, intratumoral neovascularization and tumor cell proliferation was attenuated in mice injected with Pim-3 kinase inactive cells, eventually reducing tumorigenicity in these mice to 46.6%. Moreover, Pim-3 overexpression upregulated the intratumoral levels of pSTAT3Try705, pSurvivinThr34, HGF, EGF, FGF-2 and VEGF, while the increases were markedly diminished on Pim-3 kinase inactivation. Collectively, the Pim-3 kinase emerges as being involved in accelerating human pancreatic cancer development and in promoting tumor neovascularization and subsequent tumor growth. Targeting Pim-3 may play a dual role in halting tumor progression, by promoting tumor cell death and blocking angiogenesis.

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1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2013. CA Cancer J Clin. 63:11–30. 2013. View Article : Google Scholar
2	Winter JM, Cameron JL, Campbell KA, et al: 1423 pancreaticoduodenectomies for pancreatic cancer: a single-institution experience. J Gastrointest Surg. 10:1199–1211. 2006. View Article : Google Scholar : PubMed/NCBI
3	Wang Z, Li Y, Ahmad A, et al: Pancreatic cancer: understanding and overcoming chemoresistance. Nat Rev Gastroenterol Hepatol. 8:27–33. 2011. View Article : Google Scholar : PubMed/NCBI
4	Chu D, Kohlmann W and Adler DG: Identification and screening of individuals at increased risk for pancreatic cancer with emphasis on known environmental and genetic factors and hereditary syndromes. JOP. 11:203–212. 2010.PubMed/NCBI
5	Li D, Xie K, Wolff R and Abbruzzese JL: Pancreatic cancer. Lancet. 363:1049–1057. 2004. View Article : Google Scholar
6	Vincent A, Herman J, Schulick R, Hruban RH and Goggins M: Pancreatic cancer. Lancet. 378:607–620. 2011. View Article : Google Scholar
7	Zavoral M, Minarikova P, Zavada F, Salek C and Minarik M: Molecular biology of pancreatic cancer. World J Gastroenterol. 17:2897–2908. 2011. View Article : Google Scholar
8	Li YY, Popivanova BK, Nagai Y, Ishikura H, Fujii C and Mukaida N: Pim-3, a proto-oncogene with serine/threonine kinase activity, is aberrantly expressed in human pancreatic cancer and phosphorylates bad to block bad-mediated apoptosis in human pancreatic cancer cell lines. Cancer Res. 66:6741–6747. 2006. View Article : Google Scholar
9	Feldman JD, Vician L, Crispino M, et al: KID-1, a protein kinase induced by depolarization in brain. J Biol Chem. 273:16535–16543. 1998. View Article : Google Scholar : PubMed/NCBI
10	Deneen B, Welford SM, Ho T, Hernandez F, Kurland I and Denny CT: PIM3 proto-oncogene kinase is a common transcriptional target of divergent EWS/ETS oncoproteins. Mol Cell Biol. 23:3897–3908. 2003. View Article : Google Scholar : PubMed/NCBI
11	Fujii C, Nakamoto Y, Lu P, et al: Aberrant expression of serine/threonine kinase Pim-3 in hepatocellular carcinoma development and its role in the proliferation of human hepatoma cell lines. Int J Cancer. 114:209–218. 2005. View Article : Google Scholar : PubMed/NCBI
12	Popivanova BK, Li YY, Zheng H, et al: Proto-oncogene, Pim-3 with serine/threonine kinase activity, is aberrantly expressed in human colon cancer cells and can prevent Bad-mediated apoptosis. Cancer Sci. 98:321–328. 2007. View Article : Google Scholar
13	Zheng HC, Tsuneyama K, Takahashi H, et al: Aberrant Pim-3 expression is involved in gastric adenoma-adenocarcinoma sequence and cancer progression. J Cancer Res Clin Oncol. 134:481–488. 2008. View Article : Google Scholar : PubMed/NCBI
14	Wu Y, Wang YY, Nakamoto Y, et al: Accelerated hepatocellular carcinoma development in mice expressing the Pim-3 transgene selectively in the liver. Oncogene. 29:2228–2237. 2010. View Article : Google Scholar : PubMed/NCBI
15	Zhang F, Liu B, Wang Z, et al: A novel regulatory mechanism of Pim-3 kinase stability and its involvement in pancreatic cancer progression. Mol Cancer Res. 11:1508–1520. 2013. View Article : Google Scholar : PubMed/NCBI
16	Yunis AA, Arimura GK and Russin DJ: Human pancreatic carcinoma (MIA PaCa-2) in continuous culture: sensitivity to asparaginase. Int J Cancer. 19:128–135. 1977. View Article : Google Scholar : PubMed/NCBI
17	Yano T, Ishikura H, Kato H, et al: Vaccination effect of interleukin-6-producing pancreatic cancer cells in nude mice: a model of tumor prevention and treatment in immune-compromised patients. Jpn J Cancer Res. 92:83–87. 2001. View Article : Google Scholar : PubMed/NCBI
18	Wang YY, Taniguchi T, Baba T, Li YY, Ishibashi H and Mukaida N: Identification of a phenanthrene derivative as a potent anticancer drug with Pim kinase inhibitory activity. Cancer Sci. 103:107–115. 2012. View Article : Google Scholar : PubMed/NCBI
19	Wang C, Li HY, Liu B, Huang S, Wu L and Li YY: Pim-3 promotes the growth of human pancreatic cancer in the orthotopic nude mouse model through vascular endothelium growth factor. J Surg Res. 185:595–604. 2013. View Article : Google Scholar : PubMed/NCBI
20	Xu D, Cobb MG, Gavilano L, et al: Inhibition of oncogenic Pim-3 kinase modulates transformed growth and chemosensitizes pancreatic cancer cells to gemcitabine. Cancer Biol Ther. 14:492–501. 2013. View Article : Google Scholar : PubMed/NCBI
21	Qian KC, Wang L, Hickey ER, et al: Structural basis of constitutive activity and a unique nucleotide binding mode of human Pim-1 kinase. J Biol Chem. 280:6130–6137. 2005. View Article : Google Scholar : PubMed/NCBI
22	Yang XY, Ren CP, Wang L, et al: Identification of differentially expressed genes in metastatic and non-metastatic nasopharyngeal carcinoma cells by suppression subtractive hybridization. Cell Oncol. 27:215–223. 2005.
23	Byers LA, Sen B, Saigal B, et al: Reciprocal regulation of c-Src and STAT3 in non-small cell lung cancer. Clin Cancer Res. 15:6852–6861. 2009. View Article : Google Scholar : PubMed/NCBI
24	He M and Young CY: New approaches to target the androgen receptor and STAT3 for prostate cancer treatments. Mini Rev Med Chem. 9:395–400. 2009. View Article : Google Scholar : PubMed/NCBI
25	Kim DY, Cha ST, Ahn DH, et al: STAT3 expression in gastric cancer indicates a poor prognosis. J Gastroenterol Hepatol. 24:646–651. 2009. View Article : Google Scholar : PubMed/NCBI
26	Scholz A, Heinze S, Detjen KM, et al: Activated signal transducer and activator of transcription 3 (STAT3) supports the malignant phenotype of human pancreatic cancer. Gastroenterology. 125:891–905. 2003. View Article : Google Scholar : PubMed/NCBI
27	Hirano T, Ishihara K and Hibi M: Roles of STAT3 in mediating the cell growth, differentiation and survival signals relayed through the IL-6 family of cytokine receptors. Oncogene. 19:2548–2556. 2000. View Article : Google Scholar : PubMed/NCBI
28	Shirogane T, Fukada T, Muller JM, Shima DT, Hibi M and Hirano T: Synergistic roles for Pim-1 and c-Myc in STAT3-mediated cell cycle progression and antiapoptosis. Immunity. 11:709–719. 1999. View Article : Google Scholar : PubMed/NCBI
29	Aksoy I, Sakabedoyan C, Bourillot PY, et al: Self-renewal of murine embryonic stem cells is supported by the serine/threonine kinases Pim-1 and Pim-3. Stem Cells. 25:2996–3004. 2007. View Article : Google Scholar : PubMed/NCBI
30	Li YY, Wu Y, Tsuneyama K, Baba T and Mukaida N: Essential contribution of Ets-1 to constitutive Pim-3 expression in human pancreatic cancer cells. Cancer Sci. 100:396–404. 2009. View Article : Google Scholar : PubMed/NCBI
31	Mackenzie GG, Huang L, Alston N, et al: Targeting mitochondrial STAT3 with the novel phospho-valproic acid (MDC-1112) inhibits pancreatic cancer growth in mice. PLoS One. 8:e615322013. View Article : Google Scholar : PubMed/NCBI
32	You W, Tang Q, Zhang C, et al: IL-26 promotes the proliferation and survival of human gastric cancer cells by regulating the balance of STAT1 and STAT3 activation. PLoS One. 8:e635882013. View Article : Google Scholar : PubMed/NCBI
33	Thoennissen NH, Iwanski GB, Doan NB, et al: Cucurbitacin B induces apoptosis by inhibition of the JAK/STAT pathway and potentiates antiproliferative effects of gemcitabine on pancreatic cancer cells. Cancer Res. 69:5876–5884. 2009. View Article : Google Scholar : PubMed/NCBI
34	Chang M, Kanwar N, Feng E, et al: PIM kinase inhibitors downregulate STAT3(Try705) phosphorylation. Mol Cancer Ther. 9:2478–2487. 2010. View Article : Google Scholar : PubMed/NCBI
35	Talbot DC, Ranson M, Davies J, et al: Tumor survivin is downregulated by the antisense oligonucleotide LY2181308: a proof-of-concept, first-in-human dose study. Clin Cancer Res. 16:6150–6158. 2010. View Article : Google Scholar : PubMed/NCBI
36	Sah NK, Khan Z, Khan GJ and Bisen PS: Structural, functional and therapeutic biology of survivin. Cancer Lett. 244:164–171. 2006. View Article : Google Scholar : PubMed/NCBI
37	Liu BB and Wang WH: Survivin and pancreatic cancer. World J Clin Oncol. 2:164–168. 2011. View Article : Google Scholar : PubMed/NCBI
38	Diaz N, Minton S, Cox C, et al: Activation of stat3 in primary tumors from high-risk breast cancer patients is associated with elevated levels of activated SRC and survivin expression. Clin Cancer Res. 12:20–28. 2006. View Article : Google Scholar : PubMed/NCBI
39	Kanwar JR, Kamalapuram SK and Kanwar RK: Targeting survivin in cancer: the cell-signalling perspective. Drug Discov Today. 16:485–494. 2011. View Article : Google Scholar : PubMed/NCBI
40	O’Connor DS, Grossman D, Plescia J, et al: Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Natl Acad Sci USA. 97:13103–13107. 2000.PubMed/NCBI
41	Wheatley SP and McNeish IA: Survivin: a protein with dual roles in mitosis and apoptosis. Int Rev Cytol. 247:35–88. 2005. View Article : Google Scholar : PubMed/NCBI
42	Zhang Y, Park TS and Gidday JM: Hypoxic preconditioning protects human brain endothelium from ischemic apoptosis by Akt-dependent survivin activation. Am J Physiol Heart Circ Physiol. 292:H2573–H2581. 2007. View Article : Google Scholar : PubMed/NCBI
43	Hanahan D and Weinberg RA: Hallmarks of cancer: the next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
44	Cao Z, Shang B, Zhang G, et al: Tumor cell-mediated neovascularization and lymphangiogenesis contrive tumor progression and cancer metastasis. Biochim Biophys Acta. 1836:273–286. 2013.PubMed/NCBI
45	Folkman J: Tumor angiogenesis: therapeutic implications. N Engl J Med. 285:1182–1186. 1971. View Article : Google Scholar : PubMed/NCBI
46	Kalluri R and Zeisberg M: Fibroblasts in cancer. Nat Rev Cancer. 6:392–401. 2006. View Article : Google Scholar
47	Carmeliet P and Jain RK: Molecular mechanisms and clinical applications of angiogenesis. Nature. 473:298–307. 2011. View Article : Google Scholar : PubMed/NCBI
48	Johannessen TC, Wagner M, Straume O, Bjerkvig R and Eikesdal HP: Tumor vasculature: the Achilles’ heel of cancer? Expert Opin Ther Targets. 17:7–20. 2013.
49	De Bock K, Mazzone M and Carmeliet P: Antiangiogenic therapy, hypoxia, and metastasis: risky liaisons, or not? Nat Rev Clin Oncol. 8:393–404. 2011.