Downregulation of Sp1 is involved in β-lapachone-induced cell cycle arrest and apoptosis in oral squamous cell carcinoma

Jeon,Young-Joo; Bang,Woong; Shin,Jae-Cheon; Park,Seon-Min; Cho,Jung-Jae; Choi,Yung  Hyun; Seo,Kang  Seok; Choi,Nag-Jin; Shim,Jung-Hyun; Chae,Jung-Il

doi:10.3892/ijo.2015.2972

Downregulation of Sp1 is involved in β-lapachone-induced cell cycle arrest and apoptosis in oral squamous cell carcinoma

Authors:
- Young-Joo Jeon
- Woong Bang
- Jae-Cheon Shin
- Seon-Min Park
- Jung-Jae Cho
- Yung Hyun Choi
- Kang Seok Seo
- Nag-Jin Choi
- Jung-Hyun Shim
- Jung-Il Chae
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Affiliations: Department of Oral Pharmacology, School of Dentistry and Institute of Oral Bioscience, BK21 plus, Chonbuk National University, Jeonju 561-756, Republic of Korea, Pohang Center for Evaluation of Biomaterials, Pohang, Gyeongbuk 790‑834, Republic of Korea, Department of Pharmacy, College of Pharmacy and Natural Medicine Research Institute, Mokpo National University, Jeonnam 534-729, Republic of Korea, Department of Biochemistry, Dongeui University College of Oriental Medicine, Busan 614-052, Republic of Korea, Department of Animal Science and Technology, Sunchon National University, Suncheon, Republic of Korea, Department of Animal Science, College of Agricultural and Life Science, Chonbuk National University, Jeonju 651-756, Republic of Korea
Published online on: April 20, 2015 https://doi.org/10.3892/ijo.2015.2972
Pages: 2606-2612

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Abstract

β-lapachone (β-lap) is a naturally occurring quinone obtained from the bark of lapacho tree (Tabebuia avellanedae) with anti-proliferative properties against various cancers. The present study investigated the cell proliferation and apoptosis effect of β-lap on two oral squamous cell carcinoma lines (OSCCs). We carried out a series of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl-2H-tetrazolium (MTS) assays, 4',6-diamidino-2-phenylindole (DAPI) staining, cell cycle analysis, and western blot analysis to characterize β-lap and its underlying signaling pathway. We demonstrated that β-lap-treated cells significantly reduced cell proliferation but increased DNA condensation and increased sub-G1 population in OSCCs. Particularly, β-lap suppresses activation of transcription factor specificity protein 1 (Sp1) followed by apoptosis in a concentration-dependent manner in OSCCs. Furthermore, β-lap modulated protein expression levels of cell cycle regulatory proteins and apoptosis-related proteins that are known as Sp1 target genes, resulting in apoptosis. Our results collectively indicated that β-lap was able to modulate Sp1 transactivation and induce apoptosis through the regulation of cell cycle and apoptosis-related proteins. Therefore, β-lap may be used in cancer prevention and therapies to improve clinical outcome as an anticancer drug candidate.

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1	Forastiere A, Koch W, Trotti A and Sidransky D: Head and neck cancer. N Engl J Med. 345:1890–1900. 2001. View Article : Google Scholar
2	Jefferies S and Foulkes WD: Genetic mechanisms in squamous cell carcinoma of the head and neck. Oral Oncol. 37:115–126. 2001. View Article : Google Scholar : PubMed/NCBI
3	Herskovic A, Martz K, al-Sarraf M, Leichman L, Brindle J, Vaitkevicius V, Cooper J, Byhardt R, Davis L and Emami B: Combined chemotherapy and radiotherapy compared with radiotherapy alone in patients with cancer of the esophagus. N Engl J Med. 326:1593–1598. 1992. View Article : Google Scholar : PubMed/NCBI
4	Warnakulasuriya S: Global epidemiology of oral and oropharyngeal cancer. Oral Oncol. 45:309–316. 2009. View Article : Google Scholar
5	Moon DO, Kang CH, Kim MO, Jeon YJ, Lee JD, Choi YH and Kim GY: Beta-lapachone (LAPA) decreases cell viability and telomerase activity in leukemia cells: Suppression of telomerase activity by LAPA. J Med Food. 13:481–488. 2010. View Article : Google Scholar : PubMed/NCBI
6	Ferreira SB, Salomão K, de Carvalho da Silva F, Pinto AV, Kaiser CR, Pinto AC, Ferreira VF and de Castro SL: Synthesis and anti-Trypanosoma cruzi activity of β-lapachone analogues. Eur J Med Chem. 46:3071–3077. 2011. View Article : Google Scholar : PubMed/NCBI
7	de Almeida ER, da Silva Filho AA, dos Santos ER and Lopes CA: Antiinflammatory action of lapachol. J Ethnopharmacol. 29:239–241. 1990. View Article : Google Scholar : PubMed/NCBI
8	Tzeng HP, Ho FM, Chao KF, Kuo ML, Lin-Shiau SY and Liu SH: beta-Lapachone reduces endotoxin-induced macrophage activation and lung edema and mortality. Am J Respir Crit Care Med. 168:85–91. 2003. View Article : Google Scholar : PubMed/NCBI
9	Guiraud P, Steiman R, Campos-Takaki GM, Seigle-Murandi F and Simeon de Buochberg M: Comparison of antibacterial and antifungal activities of lapachol and beta-lapachone. Planta Med. 60:373–374. 1994. View Article : Google Scholar : PubMed/NCBI
10	Li CJ, Zhang LJ, Dezube BJ, Crumpacker CS and Pardee AB: Three inhibitors of type 1 human immunodeficiency virus long terminal repeat-directed gene expression and virus replication. Proc Natl Acad Sci USA. 90:1839–1842. 1993. View Article : Google Scholar : PubMed/NCBI
11	Kung HN, Yang MJ, Chang CF, Chau YP and Lu KS: In vitro and in vivo wound healing-promoting activities of beta-lapachone. Am J Physiol Cell Physiol. 295:C931–C943. 2008. View Article : Google Scholar : PubMed/NCBI
12	Planchon SM, Wuerzberger S, Frydman B, Witiak DT, Hutson P, Church DR, Wilding G and Boothman DA: Beta-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: A p53-independent response. Cancer Res. 55:3706–3711. 1995.PubMed/NCBI
13	Planchon SM, Pink JJ, Tagliarino C, Bornmann WG, Varnes ME and Boothman DA: beta-Lapachone-induced apoptosis in human prostate cancer cells: Involvement of NQO1/xip3. Exp Cell Res. 267:95–106. 2001. View Article : Google Scholar : PubMed/NCBI
14	Li CJ, Averboukh L and Pardee AB: beta-Lapachone, a novel DNA topoisomerase I inhibitor with a mode of action different from camptothecin. J Biol Chem. 268:22463–22468. 1993.PubMed/NCBI
15	Krishnan P and Bastow KF: Novel mechanism of cellular DNA topoisomerase II inhibition by the pyranonaphthoquinone derivatives alpha-lapachone and beta-lapachone. Cancer Chemother Pharmacol. 47:187–198. 2001. View Article : Google Scholar : PubMed/NCBI
16	Lee JH, Cheong J, Park YM and Choi YH: Down-regulation of cyclooxygenase-2 and telomerase activity by beta-lapachone in human prostate carcinoma cells. Pharmacol Res. 51:553–560. 2005. View Article : Google Scholar : PubMed/NCBI
17	Wierstra I: Sp1: emerging roles - beyond constitutive activation of TATA-less housekeeping genes. Biochem Biophys Res Commun. 372:1–13. 2008. View Article : Google Scholar : PubMed/NCBI
18	Black AR, Black JD and Azizkhan-Clifford J: Sp1 and krüppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol. 188:143–160. 2001. View Article : Google Scholar : PubMed/NCBI
19	Jiang Y, Wang L, Gong W, Wei D, Le X, Yao J, Ajani J, Abbruzzese JL, Huang S and Xie K: A high expression level of insulin-like growth factor I receptor is associated with increased expression of transcription factor Sp1 and regional lymph node metastasis of human gastric cancer. Clin Exp Metastasis. 21:755–764. 2004. View Article : Google Scholar
20	Chae JI, Jeon YJ and Shim JH: Downregulation of Sp1 is involved in honokiol-induced cell cycle arrest and apoptosis in human malignant pleural mesothelioma cells. Oncol Rep. 29:2318–2324. 2013.PubMed/NCBI
21	Kim DW, Ko SM, Jeon YJ, Noh YW, Choi NJ, Cho SD, Moon HS, Cho YS, Shin JC, Park SM, et al: Anti-proliferative effect of honokiol in oral squamous cancer through the regulation of specificity protein 1. Int J Oncol. 43:1103–1110. 2013.PubMed/NCBI
22	Mann J: Natural products in cancer chemotherapy: Past, present and future. Nat Rev Cancer. 2:143–148. 2002. View Article : Google Scholar
23	Wilson RM and Danishefsky SJ: Small molecule natural products in the discovery of therapeutic agents: The synthesis connection. J Org Chem. 71:8329–8351. 2006. View Article : Google Scholar : PubMed/NCBI
24	Jackson JK, Higo T, Hunter WL and Burt HM: Topoisomerase inhibitors as anti-arthritic agents. Inflamm Res. 57:126–134. 2008. View Article : Google Scholar : PubMed/NCBI
25	Tudan C, Jackson JK, Higo TT and Burt HM: The effect of inhibiting topoisomerase I and II on the anti-apoptotic response associated with pro-inflammatory crystals of calcium pyrophosphate dihydrate in human neutrophils. Inflamm Res. 52:8–17. 2003. View Article : Google Scholar : PubMed/NCBI
26	Pink JJ, Wuerzberger-Davis S, Tagliarino C, Planchon SM, Yang X, Froelich CJ and Boothman DA: Activation of a cysteine protease in MCF-7 and T47D breast cancer cells during beta-lapachone-mediated apoptosis. Exp Cell Res. 255:144–155. 2000. View Article : Google Scholar : PubMed/NCBI
27	Wuerzberger SM, Pink JJ, Planchon SM, Byers KL, Bornmann WG and Boothman DA: Induction of apoptosis in MCF-7:WS8 breast cancer cells by beta-lapachone. Cancer Res. 58:1876–1885. 1998.PubMed/NCBI
28	Choi YH, Kang HS and Yoo MA: Suppression of human prostate cancer cell growth by beta-lapachone via down-regulation of pRB phosphorylation and induction of Cdk inhibitor p21(WAF1/CIP1). J Biochem Mol Biol. 36:223–229. 2003. View Article : Google Scholar : PubMed/NCBI
29	Dong GZ, Oh ET, Lee H, Park MT, Song CW and Park HJ: Beta-lapachone suppresses radiation-induced activation of nuclear factor-kappaB. Exp Mol Med. 42:327–334. 2010. View Article : Google Scholar : PubMed/NCBI
30	Safe S and Abdelrahim M: Sp transcription factor family and its role in cancer. Eur J Cancer. 41:2438–2448. 2005. View Article : Google Scholar : PubMed/NCBI
31	Li L and Davie JR: The role of Sp1 and Sp3 in normal and cancer cell biology. Ann Anat. 192:275–283. 2010. View Article : Google Scholar : PubMed/NCBI
32	Hsu TI, Wang MC, Chen SY, Yeh YM, Su WC, Chang WC and Hung JJ: Sp1 expression regulates lung tumor progression. Oncogene. 31:3973–3988. 2012. View Article : Google Scholar :
33	Kavurma MM and Khachigian LM: Sp1 inhibits proliferation and induces apoptosis in vascular smooth muscle cells by repressing p21^WAF1/Cip1 transcription and cyclin D1-Cdk4-p21^WAF1/Cip1 complex formation. J Biol Chem. 278:32537–32543. 2003. View Article : Google Scholar : PubMed/NCBI
34	Xu R, Zhang P, Huang J, Ge S, Lu J and Qian G: Sp1 and Sp3 regulate basal transcription of the survivin gene. Biochem Biophys Res Commun. 356:286–292. 2007. View Article : Google Scholar : PubMed/NCBI
35	Sankpal UT, Goodison S, Abdelrahim M and Basha R: Targeting Sp1 transcription factors in prostate cancer therapy. Med Chem. 7:518–525. 2011. View Article : Google Scholar : PubMed/NCBI
36	Chae JI, Jeon YJ and Shim JH: Anti-proliferative properties of kahweol in oral squamous cancer through the regulation specificity protein 1. Phytother Res. 28:1879–1886. 2014. View Article : Google Scholar : PubMed/NCBI
37	Sherr CJ and Roberts JM: CDK inhibitors: Positive and negative regulators of G1-phase progression. Genes Dev. 13:1501–1512. 1999. View Article : Google Scholar : PubMed/NCBI
38	Murray AW: Recycling the cell cycle: Cyclins revisited. Cell. 116:221–234. 2004. View Article : Google Scholar : PubMed/NCBI
39	Sherr CJ: G1 phase progression: Cycling on cue. Cell. 79:551–555. 1994. View Article : Google Scholar : PubMed/NCBI
40	Ewen ME and Lamb J: The activities of cyclin D1 that drive tumorigenesis. Trends Mol Med. 10:158–162. 2004. View Article : Google Scholar : PubMed/NCBI
41	Weinstein IB: Relevance of cyclin D1 and other molecular markers to cancer chemoprevention. J Cell Biochem. (Suppl 25): S23–S28. 1996. View Article : Google Scholar
42	Xu Q, Liu M, Xu N and Zhu H: Variation in Sp1 binding sites correlates with expression of survivin in breast cancer. Mol Med Rep. 10:1395–1399. 2014.PubMed/NCBI