KLF5 promotes hypoxia-induced survival and inhibits apoptosis in non-small cell lung cancer cells via HIF-1α

Li,Xiaochen; Liu,Xiansheng; Xu,Yongjian; Liu,Jin; Xie,Min; Ni,Wang; Chen,Shixin

doi:10.3892/ijo.2014.2544

KLF5 promotes hypoxia-induced survival and inhibits apoptosis in non-small cell lung cancer cells via HIF-1α

Authors:
- Xiaochen Li
- Xiansheng Liu
- Yongjian Xu
- Jin Liu
- Min Xie
- Wang Ni
- Shixin Chen
View Affiliations

Affiliations: Department of Respiratory and Critical Care Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P.R. China
Published online on: July 17, 2014 https://doi.org/10.3892/ijo.2014.2544
Pages: 1507-1514

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

Transcription factor Krüppel-like factors 5 (KLF5) is overexpressed in a wide range of tumor tissues and acts as a prognostic factor in cancer. However, the role of KLF5 in non-small cell lung cancer is not clear. Hypoxia plays a vital part in the development of cancer via hypoxia-inducible factor 1 (HIF-1). Our study showed that hypoxia (1% O2) increased cell viability, clonality and proliferation and inhibited cell apoptosis in A549 cells. The expression of HIF-1α and KLF5 was increased time-dependently in hypoxia. Using small interfering RNA (siRNA) targeting KLF5 or HIF-1α, we demonstrated that KLF5 or HIF-1α knockdown inhibited hypoxia-induced cell survival and promoted cell apoptosis by actively downregulating cyclin B1, survivin and upregulating caspase-3. Given the similar effect of KLF5 and HIF-1α on cell survival, an attempt was made to investigate the putative interaction of them in hypoxia. KLF5 was revealed to co-immunoprecipitate with HIF-1α and hypoxia increased the amount of KLF5 and HIF-1α complex. Moreover, silencing of KLF5 decreased HIF-1α expression while KLF5 was not affected by HIF-1α inhibition in hypoxia, confirming the effect of KLF5 on upregulation of HIF-1α. In conclusion, this study identified hypoxia as a tumor promoter by triggering KLF5 → HIF-1α → cyclin B1/survivin/caspase-3 in lung cancer cells.

View Figures

View References

1	Estimated Incidence, Mortality and Prevalence Worldwide in 2012. //globocaniarc.fr/Pages/fact_sheets_cancer.aspx. Accessed Mar 27, 2014
2	Evan GI and Vousden KH: Proliferation, cell cycle and apoptosis in cancer. Nature. 411:342–348. 2001. View Article : Google Scholar : PubMed/NCBI
3	Wilson WR and Hay MP: Targeting hypoxia in cancer therapy. Nature reviews Cancer. 11:393–410. 2011. View Article : Google Scholar
4	Brahimi-Horn MC, Chiche J and Pouyssegur J: Hypoxia and cancer. J Mol Med. 85:1301–1307. 2007. View Article : Google Scholar
5	Zhang H, Okamoto M, Panzhinskiy E, Zawada WM and Das M: PKC delta/midkine pathway drives hypoxia-induced proliferation and differentiation of human lung epithelial cells. Am J Physiol Cell Physiol. 306:C648–C658. 2014. View Article : Google Scholar : PubMed/NCBI
6	Vaporidi K, Tsatsanis C, Georgopoulos D and Tsichlis PN: Effects of hypoxia and hypercapnia on surfactant protein expression proliferation and apoptosis in A549 alveolar epithelial cells. Life Sci. 78:284–293. 2005. View Article : Google Scholar : PubMed/NCBI
7	Zagzag D, Zhong H, Scalzitti JM, Laughner E, Simons JW and Semenza GL: Expression of hypoxia-inducible factor 1alpha in brain tumors: association with angiogenesis, invasion, and progression. Cancer. 88:2606–2618. 2000. View Article : Google Scholar : PubMed/NCBI
8	Volm M and Koomagi R: Hypoxia-inducible factor (HIF-1) and its relationship to apoptosis and proliferation in lung cancer. Anticancer Res. 20:1527–1533. 2000.PubMed/NCBI
9	Wang Q, Li LH, Gao GD, et al: HIF-1alpha up-regulates NDRG1 expression through binding to NDRG1 promoter, leading to proliferation of lung cancer A549 cells. Mol Biol Rep. 40:3723–3729. 2013. View Article : Google Scholar : PubMed/NCBI
10	Luo F, Liu X, Yan N, et al: Hypoxia-inducible transcription factor-1alpha promotes hypoxia-induced A549 apoptosis via a mechanism that involves the glycolysis pathway. BMC Cancer. 6:262006. View Article : Google Scholar : PubMed/NCBI
11	Chi JT, Wang Z, Nuyten DS, et al: Gene expression programs in response to hypoxia: cell type specificity and prognostic significance in human cancers. PLoS Med. 3:e472006. View Article : Google Scholar : PubMed/NCBI
12	Kim WY and Kaelin WG: Role of VHL gene mutation in human cancer. J Clin Oncol. 22:4991–5004. 2004. View Article : Google Scholar : PubMed/NCBI
13	Sermeus A and Michiels C: Reciprocal influence of the p53 and the hypoxic pathways. Cell Death Dis. 2:e1642011. View Article : Google Scholar : PubMed/NCBI
14	Lee SJ, No YR, Dang DT, et al: Regulation of hypoxia-inducible factor 1alpha (HIF-1alpha) by lysophosphatidic acid is dependent on interplay between p53 and Kruppel-like factor 5. J Biol Chem. 288:25244–25253. 2013. View Article : Google Scholar : PubMed/NCBI
15	Park SY, Jeong KJ, Lee J, et al: Hypoxia enhances LPA-induced HIF-1alpha and VEGF expression: their inhibition by resveratrol. Cancer Lett. 258:63–69. 2007. View Article : Google Scholar : PubMed/NCBI
16	Schumpelick VH-PB and Schackert HK: Chirurgisches Forum und DGAV Forum. Springer; Berlin: 2009
17	Meyer SE, Hasenstein JR, Baktula A, et al: Kruppel-like factor 5 is not required for K-RasG12D lung tumorigenesis, but represses ABCG2 expression and is associated with better disease-specific survival. Am J Pathol. 177:1503–1513. 2010. View Article : Google Scholar
18	Tong D, Czerwenka K, Heinze G, et al: Expression of KLF5 is a prognostic factor for disease-free survival and overall survival in patients with breast cancer. Clin Cancer Res. 12:2442–2448. 2006. View Article : Google Scholar : PubMed/NCBI
19	Budihardjo I, Oliver H, Lutter M, Luo X and Wang X: Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol. 15:269–290. 1999. View Article : Google Scholar : PubMed/NCBI
20	Ambrosini G, Adida C and Altieri DC: A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med. 3:917–921. 1997. View Article : Google Scholar : PubMed/NCBI
21	Harris AL: Hypoxia - a key regulatory factor in tumour growth. Nature reviews Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
22	Graves EE, Maity A and Le QT: The tumor microenvironment in non-small-cell lung cancer. Semin Rad Oncol. 20:156–163. 2010. View Article : Google Scholar : PubMed/NCBI
23	Cui J, Mao X, Olman V, Hastings PJ and Xu Y: Hypoxia and miscoupling between reduced energy efficiency and signaling to cell proliferation drive cancer to grow increasingly faster. J Mol Cell Biol. 4:174–176. 2012. View Article : Google Scholar : PubMed/NCBI
24	Bruick RK: Oxygen sensing in the hypoxic response pathway: regulation of the hypoxia-inducible transcription factor. Genes Dev. 17:2614–2623. 2003. View Article : Google Scholar : PubMed/NCBI
25	Lenihan CR and Taylor CT: The impact of hypoxia on cell death pathways. Biochem Soc Trans. 41:657–663. 2013. View Article : Google Scholar : PubMed/NCBI
26	Hu Y, Liu J and Huang H: Recent agents targeting HIF-1alpha for cancer therapy. J Cell Biochem. 114:498–509. 2013. View Article : Google Scholar : PubMed/NCBI
27	Diakiw SM, D’Andrea RJ and Brown AL: The double life of KLF5: opposing roles in regulation of gene-expression, cellular function, and transformation. IUBMB Life. 65:999–1011. 2013. View Article : Google Scholar : PubMed/NCBI
28	Nandan MO, Chanchevalap S, Dalton WB and Yang VW: Kruppel-like factor 5 promotes mitosis by activating the cyclin B1/Cdc2 complex during oncogenic Ras-mediated transformation. FEBS Lett. 579:4757–4762. 2005. View Article : Google Scholar : PubMed/NCBI
29	Reed JC: Dysregulation of apoptosis in cancer. J Clin Oncol. 17:2941–2953. 1999.PubMed/NCBI
30	Ghavami S, Hashemi M, Ande SR, et al: Apoptosis and cancer: mutations within caspase genes. J Med Genet. 46:497–510. 2009. View Article : Google Scholar : PubMed/NCBI
31	Krepela E, Dankova P, Moravcikova E, et al: Increased expression of inhibitor of apoptosis proteins, survivin and XIAP, in non-small cell lung carcinoma. Int J Oncol. 35:1449–1462. 2009. View Article : Google Scholar : PubMed/NCBI
32	Zhu N, Gu L, Findley HW, et al: KLF5 interacts with p53 in regulating survivin expression in acute lymphoblastic leukemia. J Biol Chem. 281:14711–14718. 2006. View Article : Google Scholar : PubMed/NCBI
33	Yun YJ, Li SH, Cho YS, Park JW and Chun YS: Survivin mediates prostate cell protection by HIF-1alpha against zinc toxicity. Prostate. 70:1179–1188. 2010. View Article : Google Scholar : PubMed/NCBI
34	Van Hoecke M, Prigent-Tessier AS, Garnier PE, et al: Evidence of HIF-1 functional binding activity to caspase-3 promoter after photothrombotic cerebral ischemia. Mol Cell Neurosci. 34:40–47. 2007.PubMed/NCBI
35	Majumder PK, Febbo PG, Bikoff R, et al: mTOR inhibition reverses Akt-dependent prostate intraepithelial neoplasia through regulation of apoptotic and HIF-1-dependent pathways. Nat Med. 10:594–601. 2004. View Article : Google Scholar : PubMed/NCBI
36	Zhong H, Chiles K, Feldser D, et al: Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res. 60:1541–1545. 2000.
37	Park JH, Lee JY, Shin DH, Jang KS, Kim HJ and Kong G: Loss of Mel-18 induces tumor angiogenesis through enhancing the activity and expression of HIF-1alpha mediated by the PTEN/PI3K/Akt pathway. Oncogene. 30:4578–4589. 2011. View Article : Google Scholar : PubMed/NCBI
38	Zhang M, Ma Q, Hu H, et al: Stem cell factor/c-kit signaling enhances invasion of pancreatic cancer cells via HIF-1alpha under normoxic condition. Cancer Lett. 303:108–117. 2011. View Article : Google Scholar : PubMed/NCBI
39	Sodhi A, Montaner S, Patel V, et al: The Kaposi’s sarcoma-associated herpes virus G protein-coupled receptor up-regulates vascular endothelial growth factor expression and secretion through mitogen-activated protein kinase and p38 pathways acting on hypoxia-inducible factor 1alpha. Cancer Res. 60:4873–4880. 2000.
40	Fukuda R, Hirota K, Fan F, Jung YD, Ellis LM and Semenza GL: Insulin-like growth factor 1 induces hypoxia-inducible factor 1-mediated vascular endothelial growth factor expression, which is dependent on MAP kinase and phosphatidylinositol 3-kinase signaling in colon cancer cells. J Biol Chem. 277:38205–38211. 2002. View Article : Google Scholar
41	Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3:721–732. 2003. View Article : Google Scholar
42	Zundel W, Schindler C, Haas-Kogan D, et al: Loss of PTEN facilitates HIF-1-mediated gene expression. Genes Dev. 14:391–396. 2000.PubMed/NCBI
43	Liu R, Zheng HQ, Zhou Z, Dong JT and Chen C: KLF5 promotes breast cell survival partially through fibroblast growth factor-binding protein 1-pERK-mediated dual specificity MKP-1 protein phosphorylation and stabilization. J Biol Chem. 284:16791–16798. 2009. View Article : Google Scholar : PubMed/NCBI
44	Schofield CJ and Ratcliffe PJ: Oxygen sensing by HIF hydroxylases. Nat Rev Mol Cell Biol. 5:343–354. 2004. View Article : Google Scholar : PubMed/NCBI
45	Forsythe JA, Jiang BH, Iyer NV, et al: Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol. 16:4604–4613. 1996.PubMed/NCBI
46	Melillo G, Musso T, Sica A, Taylor LS, Cox GW and Varesio L: A hypoxia-responsive element mediates a novel pathway of activation of the inducible nitric oxide synthase promoter. J Exp Med. 182:1683–1693. 1995. View Article : Google Scholar : PubMed/NCBI
47	Feldser D, Agani F, Iyer NV, Pak B, Ferreira G and Semenza GL: Reciprocal positive regulation of hypoxia-inducible factor 1alpha and insulin-like growth factor 2. Cancer Res. 59:3915–3918. 1999.PubMed/NCBI
48	Lofstedt T, Jogi A, Sigvardsson M, et al: Induction of ID2 expression by hypoxia-inducible factor-1: a role in dedifferentiation of hypoxic neuroblastoma cells. J Biol Chem. 279:39223–39231. 2004. View Article : Google Scholar : PubMed/NCBI
49	Yee Koh M, Spivak-Kroizman TR and Powis G: HIF-1 regulation: not so easy come, easy go. Trends Biochem Sci. 33:526–534. 2008.PubMed/NCBI
50	Zou C, Yu S, Xu Z, et al: ERRalpha augments HIF-1 signalling by directly interacting with HIF-1alpha in normoxic and hypoxic prostate cancer cells. J Pathol. 233:61–73. 2014. View Article : Google Scholar : PubMed/NCBI
51	Jung JE, Kim HS, Lee CS, et al: STAT3 inhibits the degradation of HIF-1alpha by pVHL-mediated ubiquitination. Exp Mol Med. 40:479–485. 2008. View Article : Google Scholar : PubMed/NCBI
52	Minet E, Mottet D, Michel G, et al: Hypoxia-induced activation of HIF-1: role of HIF-1alpha-Hsp90 interaction. FEBS Lett. 460:251–256. 1999. View Article : Google Scholar : PubMed/NCBI
53	Ohh M, Park CW, Ivan M, et al: Ubiquitination of hypoxia-inducible factor requires direct binding to the beta-domain of the von Hippel-Lindau protein. Nat Cell Biol. 2:423–427. 2000. View Article : Google Scholar : PubMed/NCBI
54	Mori A, Moser C, Lang SA, et al: Upregulation of Kruppel-like factor 5 in pancreatic cancer is promoted by interleukin-1beta signaling and hypoxia-inducible factor-1alpha. Mol Cancer Res. 7:1390–1398. 2009. View Article : Google Scholar : PubMed/NCBI