Hedgehog signaling pathway regulates human pancreatic cancer cell proliferation and metastasis

Hao,Kun; Tian,Xiao-Dong; Qin,Chang-Fu; Xie,Xue-Hai; Yang,Yin-Mo

doi:10.3892/or.2012.2210

Hedgehog signaling pathway regulates human pancreatic cancer cell proliferation and metastasis

Authors:
- Kun Hao
- Xiao-Dong Tian
- Chang-Fu Qin
- Xue-Hai Xie
- Yin-Mo Yang
View Affiliations

Affiliations: Department of General Surgery, Peking University First Hospital, Beijing 100034, P.R. China
Published online on: December 24, 2012 https://doi.org/10.3892/or.2012.2210
Pages: 1124-1132

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

Pancreatic cancer is one of the most aggressive and devastating malignancies. The Hedgehog (Hh) pathway has been reported to play an important role in pancreatic cancer development and progression. The aim of this study was to examine the activation of the Hh pathway in human pancreatic cancer tissue samples and pancreatic cancer cell lines, and the molecular mechanisms involved in the Hh pathway mediated effects on pancreatic cancer cell proliferation and invasion. The expression levels of Hh molecules in human pancreatic cancer tissue samples and pancreatic cancer cell lines were evaluated using RT-PCR. The role of the Hh pathway in cell proliferation and invasion was evaluated using flow cytometry, MTT, colony formation assays and transwell invasion assays, and the expression of cancer stem cell markers and epithelial-mesenchymal transition (EMT) were evaluated using flow cytometry and RT-PCR. Tumorigenicity assays were used to further investigate the role of the Hh pathway in vivo. Hh molecules were highly expressed in human pancreatic cancer tissue samples and pancreatic cancer cell lines. Inhibition of the Hh pathway notably decreased cell proliferation and induced apoptosis through inhibition of the PI3K/AKT pathway and cancer stem cells. Furthermore, inhibition of the Hh signaling pathway significantly inhibited EMT by suppressing the activation of transcription factors Snail and Slug, which are correlated with significantly reduced pancreatic cancer cell invasion, suggesting that the Hh signaling pathway is involved in early metastasis. These results indicate that activation of the Hh pathway is a common event. Inhibition of the Hh pathway may be a potential molecular target of new therapeutic strategies for pancreatic cancer.

View Figures

View References

1	Siegel R, Ward E, Brawley O and Jemal A: Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin. 61:212–236. 2011. View Article : Google Scholar : PubMed/NCBI
2	Magee CJ, Ghaneh P and Neoptolemos JP: Surgical and medical therapy for pancreatic carcinoma. Best Pract Res Clin Gastroenterol. 16:435–455. 2002. View Article : Google Scholar : PubMed/NCBI
3	Yeo TP, Hruban RH, Leach SD, et al: Pancreatic cancer. Curr Probl Cancer. 26:176–275. 2002. View Article : Google Scholar : PubMed/NCBI
4	Warshaw AL and Fernández-del Castillo C: Pancreatic carcinoma. N Engl J Med. 326:455–465. 1992. View Article : Google Scholar : PubMed/NCBI
5	Koorstra JB, Hustinx SR, Offerhaus GJ and Maitra A: Pancreatic carcinogenesis. Pancreatology. 8:110–125. 2008. View Article : Google Scholar
6	Mueller MT, Hermann PC, Witthauer J, Rubio-Viqueira B, Leicht SF and Huber S: Combined targeted treatment to eliminate tumorigenic cancer stem cells in human pancreatic cancer. Gastroenterology. 137:1102–1113. 2009. View Article : Google Scholar : PubMed/NCBI
7	Tanaka H, Nakamura M, Kameda C, Kubo M, Sato N and Kuroki S: The Hedgehog signaling pathway plays an essential role in maintaining the CD44⁺CD24⁻/low subpopulation and the side population of breast cancer cells. Anticancer Res. 29:2147–2157. 2009.PubMed/NCBI
8	Shankar S, Nall D, Tang SN, Meeker D, Passarini J, Sharma J and Srivastava RK: Resveratrol inhibits pancreatic cancer stem cell characteristics in human and KrasG12D transgenic mice by inhibiting pluripotency maintaining factors and epithelial-mesenchymal transition. PLoS One. 6:e165302011. View Article : Google Scholar
9	Srivastava RK, Tang SN, Zhu W, Meeker D and Shankar S: Sulforaphane synergizes with quercetin to inhibit self-renewal capacity of pancreatic cancer stem cells. Front Biosci. 3:515–528. 2011. View Article : Google Scholar : PubMed/NCBI
10	Beachy PA, Karhadkar SS and Berman DM: Tissue repair and stem cell renewal in carcinogenesis. Nature. 432:324–331. 2004. View Article : Google Scholar : PubMed/NCBI
11	Ruiz i Altaba A, Mas C and Stecca B: The Gli code: an information nexus regulating cell fate, stemness and cancer. Trends Cell Biol. 17:438–447. 2007.PubMed/NCBI
12	Ruiz i Altaba A: Therapeutic inhibition of Hedgehog-GLI signaling in cancer: epithelial, stromal, or stem cell targets? Cancer Cell. 14:281–283. 2008.PubMed/NCBI
13	Bailey JM, Singh PK and Hollingsworth MA: Cancer metastasis facilitated by developmental pathways: Sonic hedgehog, Notch, and bone morphogenic proteins. J Cell Biochem. 102:829–839. 2007. View Article : Google Scholar : PubMed/NCBI
14	Yang Y, Tian X, Xie X, Zhuang Y, Wu W and Wang W: Expression and regulation of hedgehog signaling pathway in pancreatic cancer. Langenbecks Arch Surg. 395:515–525. 2010. View Article : Google Scholar : PubMed/NCBI
15	Olive KP, Jacobetz MA, Davidson CJ, et al: Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer. Science. 324:1457–1461. 2009. View Article : Google Scholar : PubMed/NCBI
16	Lee CJ, Dosch J and Simeone DM: Pancreatic cancer stem cells. J Clin Oncol. 26:2806–2812. 2008. View Article : Google Scholar : PubMed/NCBI
17	Tang SN, Fu J, Nall D, Rodova M, Shankar S and Srivastava RK: Inhibition of sonic hedgehog pathway and pluripotency maintaining factors regulate human pancreatic cancer stem cell characteristics. Int J Cancer. 131:30–40. 2012. View Article : Google Scholar : PubMed/NCBI
18	Yao J, An Y, Wie JS, et al: Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines. Swiss Med Wkly. 141:w132082011.PubMed/NCBI
19	Strand MF, Wilson SR, Dembinski JL, et al: A novel synthetic smoothened antagonist transiently inhibits pancreatic adenocarcinoma xenografts in a mouse model. PLoS One. 6:e199042011. View Article : Google Scholar
20	Nüsslein-Volhard C and Wieschaus E: Mutations affecting segment number and polarity in Drosophila. Nature. 287:795–801. 1980.PubMed/NCBI
21	Hooper JE and Scott MP: Communicating with Hedgehogs. Nat Rev Mol Cell Biol. 6:306–317. 2005. View Article : Google Scholar : PubMed/NCBI
22	Kalderon D: Transducing the hedgehog signal. Cell. 103:371–374. 2000. View Article : Google Scholar
23	Kayed H, Kleeff J, Osman T, Keleg S, Büchler MW and Friess H: Hedgehog signaling in the normal and diseased pancreas. Pancreas. 32:119–129. 2006. View Article : Google Scholar : PubMed/NCBI
24	Berman DM, Karhadkar SS, Maitra A, et al: Widespread requirement for Hedgehog ligand stimulation in growth of digestive tract tumors. Nature. 425:846–851. 2003. View Article : Google Scholar : PubMed/NCBI
25	Kayed H, Kleeff J, Keleg S, et al: Indian hedgehog signaling pathway: expression and regulation in pancreatic cancer. Int J Cancer. 110:668–676. 2004. View Article : Google Scholar : PubMed/NCBI
26	Thayer SP, di Magliano MP, Heiser PW, et al: Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis. Nature. 425:851–856. 2003. View Article : Google Scholar : PubMed/NCBI
27	Gao J, Li Z, Chen Z, et al: Antisense Smo under the control of the PTCH1 promoter delivered by an adenoviral vector inhibits the growth of human pancreatic cancer. Gene Ther. 13:1587–1594. 2006. View Article : Google Scholar : PubMed/NCBI
28	Xu XF, Guo CY, Liu J, et al: Gli1 maintains cell survival by up-regulating IGFBP6 and Bcl-2 through promoter regions in parallel manner in pancreatic cancer cells. J Carcinog. 8:132009. View Article : Google Scholar : PubMed/NCBI
29	Yoo YA, Kang MH, Lee HJ, et al: Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt, EMT, and MMP-9 pathway in gastric cancer. Cancer Res. 71:7061–7070. 2011. View Article : Google Scholar : PubMed/NCBI
30	Buonamici S, Williams J, Morrissey M, et al: Interfering with resistance to smoothened antagonists by inhibition of the PI3K pathway in medulloblastoma. Sci Transl Med. 2:51ra702010.PubMed/NCBI
31	Chen X, Lingala S, Khoobyari S, Nolta J, Zern MA and Wu J: Epithelial mesenchymal transition and hedgehog signaling activation are associated with chemoresistance and invasion of hepatoma subpopulations. J Hepatol. 55:838–845. 2011. View Article : Google Scholar : PubMed/NCBI
32	Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J and Dirks PB: Identification of a cancer stem cell in human brain tumors. Cancer Res. 63:5821–5828. 2003.PubMed/NCBI
33	Di Fiore R, Santulli A, Ferrante RD, et al: Identification and expansion of human osteosarcoma-cancer-stem cells by long-term 3-aminobenzamide treatment. J Cell Physiol. 219:301–313. 2009.PubMed/NCBI
34	Tirino V, Desiderio V, d’Aquino R, et al: Detection and characterization of CD133⁺ cancer stem cells in human solid tumours. PLoS One. 3:e34692008. View Article : Google Scholar : PubMed/NCBI
35	Mahller YY, Williams JP, Baird WH, et al: Neuroblastoma cell lines contain pluripotent tumor initiating cells that are susceptible to a targeted oncolytic virus. PLoS One. 4:e42352009. View Article : Google Scholar : PubMed/NCBI
36	Pode-Shakked N, Metsuyanim S, Rom-Gross E, et al: Developmental tumourigenesis: NCAM as a putative marker for the malignant renal stem/progenitor cell population. J Cell Mol Med. 13:1792–1808. 2009. View Article : Google Scholar : PubMed/NCBI
37	Walter D, Satheesha S, Albrecht P, et al: CD133 positive embryonal rhabdomyosarcoma stem-like cell population is enriched in rhabdospheres. PLoS One. 6:e195062011. View Article : Google Scholar : PubMed/NCBI
38	Han YG, Spassky N, Romaguera-Ros M, Garcia-Verdugo JM, Aguilar A, Schneider-Maunoury S and Alvarez-Buylla A: Hedgehog signaling and primary cilia are required for the formation of adult neural stem cells. Nat Neurosci. 11:277–284. 2008. View Article : Google Scholar : PubMed/NCBI
39	Singh BN, Fu J, Srivastava RK and Shankar S: Hedgehog signaling antagonist GDC-0449 (Vismodegib) inhibits pancreatic cancer stem cell characteristics: molecular mechanisms. PLoS One. 6:e273062011. View Article : Google Scholar : PubMed/NCBI
40	Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW and Guba M: Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell. 1:313–323. 2007. View Article : Google Scholar : PubMed/NCBI
41	Dembinski JL and Krauss S: Characterization and functional analysis of a slow cycling stem cell-like subpopulation in pancreas adenocarcinoma. Clin Exp Metastasis. 26:611–623. 2009. View Article : Google Scholar : PubMed/NCBI
42	Feldmann G, Dhara S, Fendrich V, et al: Blockade of hedgehog signaling inhibits pancreatic cancer invasion and metastases: a new paradigm for combination therapy in solid cancers. Cancer Res. 67:2187–2196. 2007. View Article : Google Scholar : PubMed/NCBI
43	Maitah MY, Ali S, Ahmad A, Gadgeel S and Sarkar FH: Up-regulation of sonic hedgehog contributes to TGF-β1-induced epithelial to mesenchymal transition in NSCLC cells. PLoS One. 6:e160682011.PubMed/NCBI
44	Liao X, Siu MK, Au CW, et al: Aberrant activation of hedgehog signaling pathway in ovarian cancers: effect on prognosis, cell invasion and differentiation. Carcinogenesis. 30:131–140. 2009. View Article : Google Scholar : PubMed/NCBI
45	Olmeda D, Jordá M, Peinado H, Fabra A and Cano A: Snail silencing effectively suppresses tumour growth and invasiveness. Oncogene. 26:1862–1874. 2007. View Article : Google Scholar : PubMed/NCBI
46	Peinado H, Olmeda D and Cano A: Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer. 7:415–428. 2007. View Article : Google Scholar : PubMed/NCBI