[an error occurred while processing this directive]

Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (7): 645-654.doi: 10.3736/jcim20100708

• Original Experimental Research • Previous Articles     Next Articles

Anti-oncogenic potentials of a plant coumarin (7-hydroxy-6-methoxy coumarin) against 7,12-dimethylbenz[a]anthracene-induced skin papilloma in mice:The possible role of several key signal proteins

Soumya Sundar Bhattacharyyaa,Saili Paula,Suman Duttaa,Naoual Boujedainib,Anisur Rahman Khuda- Bukhsha   

  1. a Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani，Kaslyani 741235, India
    b Boiron Laboratory, 20 rue de la Liberation, Sainte-Les-Foy-Lyon 69110, France
  • Received:2010-01-25 Online:2010-07-20 Published:2018-10-20
  • Contact: Rahman Khuda- Bukhsh Anisur E-mail:khudabukhsh_48@rediffmail.com

RichHTML

5

PDF (PC)

507

Like

Objective

Anti-cancer potentials of scopoletin (7-hydroxy-6-methoxy coumarin) separated from plant extract (Gelsemium sempervirens) were demonstrated earlier from our in vitro studies. In the present study, its in vivo effects have been evaluated in mice.
Methods

Mice were chronically administered 7,12-dimethylbenz[a]anthracene (DMBA) once a week and croton oil twice a week on their back, which resulted in the development of fully grown finger-like projections (papilloma) after 24 weeks. Two subgroups of mice (drug-treated) were treated with two doses of scopoletin (50 mg and 100 mg/kg body weight) respectively while control received 2% ethyl alcohol (the “vehicle” of scopoletin). After the 24-week drug administration, expressions of several key receptors such as aryl hydrocarbon receptor (AhR) and signal proteins like p53, cytochrome P450 1A1 (CYP1A1), proliferating cell nuclear antigen (PCNA), signal transducer and activator of transcription-3 (Stat-3), survivin, matrix metalloproteinase-2 (MMP-2), cyclin D1, c-myc, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and caspase-3, and some anti-oxidant markers were studied. Lipid peroxidation, superoxide dismutase, catalase, glutathione peroxidase and glutathione-s-transferase in supernatant were also detected.
Results

Carcinogens induced toxicity, and over-expression of AhR, CYP1A1, PCNA, Stat-3, survivin, MMP-2, cyclin D1 and c-myc and down-regulation of p53, caspase-3 and TIMP-2. In mice treated with scopoletin, the expressions of these proteins and toxicity biomarkers were reverted.
Conclusion

Since AhR is known to be ligand-activated by DMBA to release signals for several downstream proteins initiating reactive oxygen species generation, the down-regulation of AhR by scopoletin appeared to play a significant role in subsequent down-regulation of some key signal proteins. One possible mechanism of down-regulation of AhR may be through competitive inhibition by scopoletin. Mitogen-activated protein kinases may also have some critical role. This compound can be considered as a possible candidate for chemoprevention.

Key words: Scopoletin, Skin neoplasms, 7, 12-dimethylbenz[a]anthracene, Signaling receptors, Apoptosis, Mice

Figure 1

Structure of scopoletin"

"

Figure 2

Histology of skin tested by haematoxylin and eosin staining and PCNA protein localization Histology of skin: A, B, and C; PCNA localization: D, E, and F. A and D: Normal group; B and E: DMBA plus croton oil group; C and F: Scopoletin-fed group. G: Proportion of average fluorescence intensity of PCNA protein in different groups. **P<0.01, vs DMBA plus alcohol group."

"

"

"

"

"

Figure 3

Immunoblots and mRNA expressions of β-actin, AhR, CYP1A1, Stat-3, p53, PCNA and caspase-3 A: Normal group; B: DMBA plus croton oil group; C: DMBA, croton oil plus alcohol group; D: DMBA, croton oil plus scopoletin (50 mg/kg) group; E: DMBA, croton oil plus scopoletin (100 mg/kg) group."

Figure 4

mRNA expressions and densitometric representation of survivin, cyclin D1, c-myc, MMP-2 and TIMP-2 A: Normal group; B: DMBA plus croton oil group; C: DMBA, croton oil plus alcohol group; D: DMBA, croton oil plus scopoletin (50 mg/kg) group; E: DMBA, croton oil plus scopoletin (100 mg/kg) group."

Figure 5

Immunoblots of expressions of NF-κB and p38MAPK A: Normal group; B: DMBA plus croton oil group; C: DMBA, croton oil plus alcohol group; D: DMBA, croton oil plus scopoletin (50 mg/kg) group; E: DMBA, croton oil plus scopoletin (100 mg/kg) group."

Figure 6

Immunoprecipitation of ARNT blots A: Normal group; B: DMBA plus croton oil group; C: DMBA, croton oil plus alcohol group; D: DMBA, croton oil plus scopoletin (50 mg/kg) group; E: DMBA, croton oil plus scopoletin (100 mg/kg) group."

[1] Kawaii S, Tomono Y, Ogawa K, Sugiura M, Yano M, Yoshizawa Y . The antiproliferative effect of coumarins on several cancer cell lines[J]. Anticancer Res, 2001,21(2A):917-923
[2] Bhattacharyya SS, Mandal SK, Biswas R, Paul S, Pathak S, Boujaedaini N, Belon P, Khuda-Bukhsh AR . In vitro studies demonstrate anticancer activity of an alkaloid of plant Gelsemium semipervirens[J]. Exp Biol Med (Maywood), 2008,233(12):1591-1601
doi: 10.3181/0805-RM-181
[3] Bhattacharyya SS, Paul S, Mondal SK, Banerjee A, Boujedaini N, Khuda-Bukhsh AR . A synthetic coumarin(4-methyl-7-hydroxy coumarin) has anti-cancer potentials against DMBA-induced skin cancer in mice[J]. Eur J Pharmacol, 2009,614(1-3):128-136
doi: 10.1016/j.ejphar.2009.04.015
[4] Takayama H, Sakai SI. Gelsemium alkaloids. In: Cordell GA. The alkaloids(Vol 49)[M]. San Diego, California: Academic Press, 1997,( Vol 49):1-78
[5] Evans WC.Trease and Evans pharmacognosy(15th ed)[M].Philadelphia: WB Saunders,2002,(15th ed)
[6] Das RK, Hossain SK, Bhattacharya S . Diphenylmethyl selenocyanate inhibits DMBA-croton oil induced two-stage skin carcinogenesis by inducing apoptosis and inhibiting cutaneous cell proliferation[J]. Cancer Lett, 2005,230(1):90-101
doi: 10.1016/j.canlet.2004.12.021
[7] Katiyar SK, Roy AM, Baliga MS . Silymarin induces apoptosis primarily through a p53-dependent pathway involving Bcl-2/Bax, cytochrome c release and caspase activation[J]. Mol Cancer Ther, 2005,4(2):207-216
[8] Motiwale L, Ingle AD, Rao KV . Mouse skin tumor promotion by sodium arsenate is associated with enhanced PCNA expression[J]. Cancer Lett, 2005,223(1):27-35
doi: 10.1016/j.canlet.2004.10.020 pmid: 15890234
[9] Limtrakul P, Anuchareeda S, Lipigorngoson S, Dunn FW . Inhibition of carcinogen induced c-Ha-ras and c-fos proto-oncogenes expression by dietary curcumin[J]. BMC Cancer, 2001,1(1):1-7
doi: 10.1186/1471-2407-1-1
[10] Yokozawa T, Chen CP, Dong E, Tanaka T, Nonaka GI, Nishioka I . Study on the inhibitory effect of tannins and flavonoids against the 1,1-diphenyl-2-picrylhydrazyl radical[J]. Biochem Pharmacol, 1998,56(2):213-222
doi: 10.1016/S0006-2952(98)00128-2
[11] Sreejayan N, Rao MNA . Nitric oxide scavenging by curcuminioids[J]. J Pharm Pharmacol, 1997,49(1):105-107
doi: 10.1111/j.2042-7158.1997.tb06761.x pmid: 9120760
[12] Mary NK, Shylesh BS, Babu BH, Padikkala J . Antioxidant and hypolipidaemic activity of a herbal formulation — liposem[J]. Ind J Exp Biol, 2002,40(8):901-904
pmid: 12597019
[13] Currier N, Solomon SE, Demicco EG, Chang DL, Farago M, Ying H, Dominguez I, Sonenshein GE, Cardiff RD, Xiao ZX, Sherr DH, Seldin DC . Oncogenic signaling pathways activated in DMBA-induced mouse mammary tumors[J]. Toxicol Pathol, 2005,33(6):726-737
doi: 10.1080/01926230500352226
[14] Ohkawa H, Ohishi N, Yagi K . Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction[J]. Anal Biochem, 1979,95(2):351-358
doi: 10.1016/0003-2697(79)90738-3 pmid: 36810
[15] Kakkar P, Das B, Viswanathan PN . A modified spectrophotometric assay of superoxide dismutase[J]. Indian J Biochem Biophys, 1984,21(2):130-132
doi: 10.1515/bchm2.1984.365.2.1489 pmid: 6490072
[16] Maehly AC, Chance B.The assay of catalase and peroxidase[J].Methods Biochem Anal , 1954, 1: 357-424
doi: 10.1002/9780470110171.ch14
[17] Paglia DE, Valentine WN . Studies on the quantitative and qualitative characterisation of erythrocyte glutathione peroxidase[J]. J Lab Clin Med, 1967,70(1):158-169
[18] Habig WH, Pabst MJ, Jakoby WB . Glutathione-S-transferase, the first enzymatic step in mercapturic acid formation[J]. J Biol Chem, 1974,249(22):7130-7139
[19] Arabzadeh A, Troy TC , Turksen K.Changes in the distribution pattern of Claudin tight junction proteins during the progression of mouse skin tumorigenesis[J].BMC Cancer, 2007, 7: 196
doi: 10.1186/1471-2407-7-196
[20] Burnette WN . “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate -polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A[J]. Anal Biochem, 1981,112(2):195-203
doi: 10.1016/0003-2697(81)90281-5
[21] Symonds H, Krall L, Remington L, Saenz-Robles M, Lowe S, Jacks T , Van Dyke T.p53-dependent apoptosis suppresses tumor growth and progression in vivo[J]. Cell, 1994,78(4):703-711
doi: 10.1016/0092-8674(94)90534-7 pmid: 8069917
[22] Lowe SW, Jacks T, Housman DE, Ruley HE . Abrogation of oncogene-associated apoptosis allows transformation of p53-deficient cells[J]. Proc Natl Acad Sci, 1994,91(6):2026-2030
doi: 10.1016/j.ajpath.2012.08.025 pmid: 8134344
[23] Donehower LA, Harvey M, Slagle BL , McArthur MJ, Montgomery CA Jr, Butel JS, Bradley A.Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumors[J]. Nature, 1992,356(6366):215-221
doi: 10.1038/356215a0 pmid: 1552940
[24] Hollstein M, Rice K, Greenblatt MS, Soussi T, Fuchs R, Sorlie T, Hovig E, Smith-Shorensen B, Montesano R, Harris CC . Database of p53 gene somatic mutations in human tumors and cell lines[J]. Nucleic Acids Res, 1994,22(17):3551-3555
[25] Venkatarachalam S, Shi YP, Jones SN, Vogel H, Bradley A, Pinkel D, Donehower LA . Retention of wild-type p53 in tumors from p53 heterozygous mice: reduction of p53 dosage can promote cancer formation[J]. EMBO J, 1998,17(16):4657-4667
doi: 10.1093/emboj/17.16.4657
[26] Li FZ, Brattain MG . Role of the survivin gene in pathophysiology[J]. Am J Pathol, 2006,169(1):1-11
doi: 10.2353/ajpath.2006.060121 pmid: 16816356
[27] Singhal R, Badger TM, Ronis MJ . Reduction in 7,12-dimethylbenz[a]anthracene-induced hepatic cytochrome-P450 1A1 expression following soy consumption in female rats is mediated by degradation of Aryl hydrocarbon receptor[J]. J Nutr, 2007,137(1):19-24
doi: 10.1093/jn/137.1.19
[28] Jung KJ, Wallig MA, Singletary KW . Purple grape juice inhibits 7,12-dimethylbenz[a]antracene(DMBA)-induced rat mammary tumorigenesis and in vivo DMBA-DNA adduct formation[J]. Cancer Lett, 2006,233(2):279-288
doi: 10.1016/j.canlet.2005.03.020
[29] Lee BC, Lee SY, Lee HJ, Sim GS, Kim JH, Kim JH, Cho YH, Lee DH, Pyo HB, Choe TB, Moon DC, Yun YP, Hong JT . Anti-oxidative and photo-protective effects of coumarins isolated from Fraxinus Chinensis[J]. Arch Pharm Res, 2007,30(10):1293-1301
doi: 10.1007/BF02980270
[30] Kim SO, Kundu JK, Shin YK, Park JH, Cho MH, Kim TY, Surh YJ .[ 6]-Gingerol inhibits COX-2 expression by blocking the activation of p38 MAP kinase and NF-kappaB in phorbol ester-stimulated mouse skin[J]. Oncogene, 2005,24(15):2558-2567
doi: 10.1038/sj.onc.1208446
[1] Bikash Manandhar, Hyun Jung Kim, Dong Young Rhyu. Caulerpa okamurae extract attenuates inflammatory interaction, regulates glucose metabolism and increases insulin sensitivity in 3T3-L1 adipocytes and RAW 264.7 macrophages. Journal of Integrative Medicine, 2020, 18(3): 253-264.
[2] Mehranghiz Ebrahimi-Mameghani, Mohammad Asghari-Jafarabadi, Khatereh Rezazadeh. TCF7L2-rs7903146 polymorphism modulates the effect of artichoke leaf extract supplementation on insulin resistance in metabolic syndrome: A randomized, double-blind, placebo-controlled trial. Journal of Integrative Medicine, 2018, 16(5): 329-334.
[3] Sandeep Kumar Singh, Arjun Patra. Evaluation of phenolic composition, antioxidant, anti-inflammatory and anticancer activities of Polygonatum verticillatum (L.). Journal of Integrative Medicine, 2018, 16(4): 273-282.
[4] Shui-jie Shen, Yong-hong Zhang, Xiao-xia Gu, Shui-ju Jiang, Ling-jun Xu. Yangfei Kongliu Formula, a compound Chinese herbal medicine, combined with cisplatin, inhibits growth of lung cancer cells through transforming growth factor-β1 signaling pathway. Journal of Integrative Medicine, 2017, 15(3): 242-251.
[5] Bhesh Raj Sharma, Dong Wook Kim, Dong Young Rhyu. Korean Chungtaejeon tea extract attenuates weight gain in C57BL/6J-Lep ob/ob mice and regulates adipogenesis and lipolysis in 3T3-L1 adipocytes. Journal of Integrative Medicine, 2017, 15(1): 56-63.
[6] Avijit Paul, Kausik Bishayee, Samrat Ghosh, Avinaba Mukherjee, Sourav Sikdar, Debrup Chakraborty, Naoual Boujedaini, Anisur Rahman Khuda-Bukhsh. Chelidonine isolated from ethanolic extract of Chelidonium majus promotes apoptosis in HeLa cells through p38-p53 and PI3K/AKT signalling pathways. Journal of Chinese Integrative Medicine, 2012, 10(9): 1025-1038.
[7] Yi-tao Zhao , Xin-guang Zhang , Li Bai , Li-qing Li , Jian-er Yu . Effects of Chinese herbal medicine Pingchuan Formula on airway inflammation, interferon-γ and interleukin-4 in mice with asthma. Journal of Chinese Integrative Medicine, 2012, 10(7): 807-813.
[8] Lin Wang, Guan-liang Wang, Jia-han Liu, Di Li, De-zeng Zhu, Liang-neng Wu . Effects of ursolic acid in ameliorating insulin resistance in liver of KKAy mice via peroxisome proliferator-activated receptors α and γ. Journal of Chinese Integrative Medicine, 2012, 10(7): 793-799.
[9] Mei-jiao Mao, Jun-ping Hu, Cong Wang, Yi-yi Zhang, Ping Liu. Effects of Chinese herbal medicine Guanxinkang on expression of PPARγ-LXRα-ABCA1 pathway in ApoE-knockout mice with atherosclerosis. Journal of Chinese Integrative Medicine, 2012, 10(7): 814-820.
[10] Dong-mei Zhang, Li-xia Lou, Ai-ming Wu, Xi-ying Lü, Zhao-juan Hu, Yan-hong Zhang, Hong-fang Liu. Effects of Astragalus membranaceus and Potentilla discolor mixture on insulin resistance and its related mRNA expressions in KKAy mice with type 2 diabetes. Journal of Chinese Integrative Medicine, 2012, 10(7): 821-826.
[11] Yogesh Bendale, Vineeta Bendale, Saili Paul, Soumya Sundar Bhattacharyya. Green synthesis, characterization and anticancer potential of platinum nanoparticles Bioplatin. Journal of Chinese Integrative Medicine, 2012, 10(6): 681-689.
[12] Surendra Kr. Sharma, Naveen Goyal. Protective effect of Heliotropium eichwaldi against cisplatin-induced nephrotoxicity in mice. Journal of Chinese Integrative Medicine, 2012, 10(5): 555-560.
[13] Jing Guo, Ju-yong Wang, Zhan Zheng, Qing Wang, Chang-sheng Dong . Effects of Chinese herbal medicine Feiyanning Decoction on the ratio of CD4+CD25+ regulatory T cells and expression of transcription factor Foxp3 in mice bearing Lewis lung carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(5): 584-590.
[14] Santu Kumar Saha, Sreemanti Das, Anisur Rahman Khuda-Bukhsh. Phenotypic evidence of ultra-highly diluted homeopathic remedies acting at gene expression level: A novel probe on experimental phage infectivity in bacteria. Journal of Chinese Integrative Medicine, 2012, 10(4): 462-470.
[15] Zhan gheng, Ju-yong Wang, Qing Wang, Ling Xu, Zhen-ye Xu. Effects of Chinese herbal medicine Feiyanning decoction on expressions of nucleosome conformation-regulating factors H3-K56, Rtt109, Asf1 and E2F1 in Lewis-bearing mice. Journal of Chinese Integrative Medicine, 2012, 10(4): 448-453.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278
[an error occurred while processing this directive]