Triptolide suppresses proliferation, hypoxia-inducible factor-1α and c-Myc expression in pancreatic cancer cells

Ding,Xiaoling; Zhou,Xiaorong; Jiang,Bo; Zhao,Qun; Zhou,Guoxiong

doi:10.3892/mmr.2015.3960

Triptolide suppresses proliferation, hypoxia-inducible factor-1α and c-Myc expression in pancreatic cancer cells

Authors:
- Xiaoling Ding
- Xiaorong Zhou
- Bo Jiang
- Qun Zhao
- Guoxiong Zhou
View Affiliations

Affiliations: Department of Gastroenterology, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China, Department of Immunology, Medical School of Nantong University, Nantong, Jiangsu 226001, P.R. China
Published online on: June 18, 2015 https://doi.org/10.3892/mmr.2015.3960
Pages: 4508-4513

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

Triptolide (TL) is known to suppress the proliferation of a number of pancreatic cancer cell lines in vitro. Marked antitumor effects were also observed in a xenograft model of pancreatic cancer. Hypoxia‑inducible factor‑1α (HIF‑1α) is highly expressed in pancreatic cancer cells lines. The present study therefore tested the hypothesis that suppression of HIF‑1α is associated with the antitumor activity of TL. Quantitative polymerase chain reaction and western blot analysis were used to determine the level of gene expression. A xenograft tumor model of pancreatic cancer was established in athymic nude mice and the tumor size was measured to evaluate the outcome of TL treatment. Immunohistochemistry was used to detect the expression of HIF‑1α and vascular endothelial growth factor (VEGF), and to assess microvessel density. Microarray was used to investigate gene expression in pancreatic cancer cells following TL treatment. The expression of HIF‑1α was shown to be reduced in pancreatic cell lines following treatment with TL, and this effect occurred in a dose‑dependent manner. In a xenograft model of pancreatic cancer, reduced levels of HIF‑1α were also observed in mice that were treated with TL. Furthermore, the expression of VEGF, which is a direct target of HIF‑1α, was also suppressed, and the microvessel density of tumor tissues was consequently reduced. A microarray analysis of gene expression was performed in order to investigate the potential mechanisms underlying the antitumor activity of TL. The results showed that 11 genes, including c‑Myc, SOX9 and Ets2, were downregulated at an early stage following treatment with TL. A recent study indicated that overexpression of c‑Myc in colon cancer cells promotes increased expression of HIF‑1α and VEGF. Therefore, TL may suppress HIF‑1α through a c‑Myc‑dependent mechanism, which is involved in antitumor effects in mouse models of pancreatic cancer.

View Figures

View References

1	Hidalgo M: Pancreatic cancer. N Engl J Med. 362:1605–1617. 2010. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
3	Castellanos E, Berlin J and Cardin DB: Current treatment options for pancreatic carcinoma. Curr Oncol Rep. 13:195–205. 2011. View Article : Google Scholar : PubMed/NCBI
4	Maitra A and Hruban RH: Pancreatic cancer. Annu Rev Pathol. 3:157–188. 2008. View Article : Google Scholar
5	Zhou GX, Ding XL, Huang JF, Zhang H, Wu SB, Cheng JP and Wei Q: Apoptosis of human pancreatic cancer cells induced by Triptolide. World J Gastroentero. 14:1504–1509. 2008. View Article : Google Scholar
6	Carter BZ, Mak DH, Schober WD, Dietrich MF, et al: Triptolide sensitizes AML cells to TRAIL-induced apoptosis via decrease of XIAP and p53-mediated increase of DR5. Blood. 111:3742–3750. 2008. View Article : Google Scholar : PubMed/NCBI
7	Chang WT, Kang JJ, Lee KY, et al: Triptolide and chemotherapy cooperate in tumor cell apoptosis. A role for the p53 pathway. J Biol Chem. 276:2221–2227. 2001. View Article : Google Scholar
8	Kulke MH: Systemic therapy for advanced pancreatic neuroendocrine tumors. Semin Oncol. 40:75–83. 2013. View Article : Google Scholar : PubMed/NCBI
9	Otrock ZK, Hatoum HA, Awada AH, Ishak RS and Shamseddine AI: Hypoxia-inducible factor in cancer angiogenesis: structure, regulation and clinical perspectives. Crit Rev Oncol Hematol. 70:93–102. 2009. View Article : Google Scholar : PubMed/NCBI
10	Semenza GL: HIF-1: upstream and downstream of cancer metabolism. Curr Opin Genet Dev. 20:51–56. 2010. View Article : Google Scholar :
11	Ding X, Zhu C, Qiang H, Zhou X and Zhou G: Enhancing antitumor effects in pancreatic cancer cells by combined use of COX-2 and 5-LOX inhibitors. Biomed Pharmacother. 65:486–490. 2011. View Article : Google Scholar : PubMed/NCBI
12	Weidner N: Intratumor microvessel density as a prognostic factor in cancer. Am J Pathol. 147:9–19. 1995.PubMed/NCBI
13	Hoffmann AC, Mori R, Vallbohmer D, et al: High expression of HIF1a is a predictor of clinical outcome in patients with pancreatic ductal adenocarcinomas and correlated to PDGFA, VEGF, and bFGF. Neoplasia. 10:674–679. 2008. View Article : Google Scholar : PubMed/NCBI
14	Sun HC, Qiu ZJ, Liu J, et al: Expression of hypoxia-inducible factor-1 alpha and associated proteins in pancreatic ductal adenocarcinoma and their impact on prognosis. Int J Oncol. 30:1359–1367. 2007.PubMed/NCBI
15	Tanaka T, Kuroki T, Adachi T, et al: Evaluation of SOX9 expression in pancreatic ductal adenocarcinoma and intraductal papillary mucinous neoplasm. Pancreas. 42:488–493. 2013. View Article : Google Scholar
16	Gao C, Xie R, Ren C and Yang X: Dickkopf-1 expression is a novel prognostic marker for gastric cancer. J Biomed Biotechnol. 2012:8045922012. View Article : Google Scholar : PubMed/NCBI
17	Lee SH, Hong HS, Liu ZX, Kim RH, Kang MK, Park NH and Shin KH: TNFα enhances cancer stem cell-like phenotype via Notch-Hes1 activation in oral squamous cell carcinoma cells. Biochem Biophys Res Commun. 424:58–64. 2012. View Article : Google Scholar : PubMed/NCBI
18	Dang CV: MYC on the path to cancer. Cell. 149:22–35. 2012. View Article : Google Scholar : PubMed/NCBI
19	Gabay M, Li Y and Felsher DW: MYC activation is a hallmark of cancer initiation and maintenance. Cold Spring Harb Perspect Med. 4:a0142412014. View Article : Google Scholar : PubMed/NCBI
20	Stellas D, Szabolcs M, Koul S, et al: Therapeutic effects of an anti-Myc drug on mouse pancreatic cancer. J Natl Cancer Inst. 106:dju3202014. View Article : Google Scholar : PubMed/NCBI
21	Zhou ZL, Yang YX, Ding J, Li YC and Miao ZH: Triptolide: structural modifications, structure-activity relationships, bioactivities, clinical development and mechanisms. Nat Prod Rep. 29:457–475. 2012. View Article : Google Scholar : PubMed/NCBI
22	Alsaied OA, Sangwan V, Banerjee S, Krosch TC, Chugh R, Saluja A, Vickers SM and Jensen EH: Sorafenib and triptolide as combination therapy for hepatocellular carcinoma. Surgery. 156:270–279. 2014. View Article : Google Scholar : PubMed/NCBI
23	Carter BZ, Mak DH, Shi Y, Fidler JM, Chen R, Ling X, Plunkett W and Andreeff M: MRx102, a triptolide derivative, has potent antileukemic activity in vitro and in a murine model of AML. Leukemia. 26:443–450. 2012. View Article : Google Scholar
24	Ding X, Zhang B, Pei Q, Pan J, Huang S, Yang Y, Zhu Z, Lv Y and Zou X: Triptolide induces apoptotic cell death of human cholangiocarcinoma cells through inhibition of myeloid cell leukemia-1. BMC Cancer. 14:2712014. View Article : Google Scholar : PubMed/NCBI
25	Liu Q: Triptolide and its expanding multiple pharmacological functions. Int Immunopharmacol. 11:377–383. 2011. View Article : Google Scholar : PubMed/NCBI
26	Yinjun L, Jie J and Yungui W: Triptolide inhibits transcription factor NF-kappaB and induces apoptosis of multiple myeloma cells. Leuk Res. 29:99–105. 2005. View Article : Google Scholar
27	Titov DV, Gilman B, He QL, Bhat S, Low WK, Dang Y, Smeaton M, Demain AL, Miller PS, Kugel JF, et al: XPB, a subunit of TFIIH, is a target of the natural product triptolide. Nat Chem Biol. 7:182–188. 2011. View Article : Google Scholar : PubMed/NCBI
28	Vispé S, DeVries L, Créancier L, Besse J, Bréand S, Hobson DJ, Svejstrup JQ, Annereau JP, Cussac D, Dumontet C, et al: Triptolide is an inhibitor of RNA polymerase I and II-dependent transcription leading predominantly to down-regulation of short-lived mRNA. Mol Cancer Ther. 8:2780–2790. 2009. View Article : Google Scholar : PubMed/NCBI
29	Wang Y, Lu JJ, He L and Yu Q: Triptolide (TPL) inhibits global transcription by inducing proteasome-dependent degradation of RNA polymerase II (Pol II). PLoS One. 6:e239932011. View Article : Google Scholar : PubMed/NCBI
30	Meyer N and Penn LZ: Reflecting on 25 years with MYC. Nat Rev Cancer. 8:976–990. 2008. View Article : Google Scholar : PubMed/NCBI
31	Rankin EB and Giaccia AJ: The role of hypoxia-inducible factors in tumorigenesis. Cell Death Differ. 15:678–685. 2008. View Article : Google Scholar : PubMed/NCBI
32	Chen C, Cai S, Wang G, Cao X, Yang X, Luo X, Feng Y and Hu J: c-Myc enhances colon cancer cell-mediated angiogenesis through the regulation of HIF-1α. Biochem Biophys Res Commun. 430:505–511. 2013. View Article : Google Scholar
33	Gordan JD, Thompson CB and Simon MC: HIF and c-Myc: sibling rivals for control of cancer cell metabolism and proliferation. Cancer Cell. 12:108–113. 2007. View Article : Google Scholar : PubMed/NCBI
34	Soucek L, Whitfield JR, Sodir NM, Massó-Vallés D, Serrano E, Karnezis AN, Swigart LB and Evan GI: Inhibition of Myc family proteins eradicates KRas-driven lung cancer in mice. Genes Dev. 27:504–513. 2013. View Article : Google Scholar : PubMed/NCBI
35	Soucek L, Whitfield J, Martins CP, Finch AJ, Murphy DJ, Sodir NM, Karnezis AN, Swigart LB, Nasi S and Evan GI: Modelling Myc inhibition as a cancer therapy. Nature. 455:679–683. 2008. View Article : Google Scholar : PubMed/NCBI
36	Sodir NM, Swigart LB, Karnezis AN, Hanahan D, Evan GI and Soucek L: Endogenous Myc maintains the tumor microenvironment. Genes Dev. 25:907–916. 2011. View Article : Google Scholar : PubMed/NCBI