Tags

Type your tag names separated by a space and hit enter

Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer.
BMC Complement Altern Med. 2017 Jan 10; 17(1):36.BC

Abstract

BACKGROUND

Previous studies in our laboratory showed that the Lebanese Daucus carota ssp. carota (wild carrot) oil extract possesses in vitro and in vivo anticancer activities. The present study aims to examine the cytotoxic effect of Daucus carota oil fractions on human epidermal keratinocytes and evaluate the chemopreventive activity of the pentane diethyl ether fraction on DMBA/TPA induced skin carcinogenesis in mice.

METHODS

Wild carrot oil extract was chromatographed to yield four fractions (F1, 100% pentane; F2, 50:50 pentane:diethyl ether; F3, 100% diethyl ether; F4 93:7 chloroform:methanol). The cytotoxic effect of fractions (10, 25, 50 and 100 μg/mL) was tested on human epidermal keratinocytes (non-tumorigenic HaCaT cells and tumorigenic HaCaT-ras variants) using WST a ssay. Cell cycle phase distribution of tumorigenic HaCaT-ras variants was determined by flow cytometry post-treatment with F2 fraction. Apoptosis related proteins were also assessed using western blot. The antitumor activity of F2 fraction was also evaluated using a DMBA/TPA induced skin carcinoma in Balb/c mice.

RESULTS

All fractions exhibited significant cytotoxicity, with HaCaT cells being 2.4-3 times less sensitive than HaCaT-ras A5 (benign tumorigenic), and HaCaT-ras II4 (malignant) cells. GC-MS analysis revealed the presence of a major compound (around 60%) in the pentane/diethylether fraction (F2), identified as 2-himachalen-6-ol. Treatment of HaCaT-ras A5 and HaCaT-ras II4 cells with F2 fraction resulted in the accumulation of cells in the sub-G1 apoptotic phase and decreased the population of cells in the S and G2/M phases. Additionally, F2 fraction treatment caused an up-regulation of the expression of pro-apoptotic (Bax) and down-regulation of the expression of anti-apoptotic (Bcl2) proteins. A decrease in the phosphorylation of AKT and ERK was also observed. Intraperitoneal treatment with F2 fraction (50 or 200 mg/kg) in the DMBA/TPA skin carcinogenesis mouse model showed a significant inhibition of papilloma incidence (mice with papilloma), yield (number of papilloma/mouse) and volume (tumor relative size) at weeks 15, 18 and 21.

CONCLUSION

The present data reveal that F2 fraction has a remarkable antitumor activity against DMBA/TPA-induced skin carcinogenesis, an effect that may be mediated through inhibition of the MAPK/ERK and PI3K/AKT pathways.

Authors+Show Affiliations

Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, P.O. Box 36, Byblos, Lebanon.School of Pharmacy, Department of Pharmaceutical Sciences, Lebanese American University, P.O. Box 36, Byblos, Lebanon.Deutsches Krebsforschungszentrum DKFZ, German Cancer Research Center, Genetics of Skin Carcinogenesis, Im Neuenheimer Feld 280, Heidelberg, 69120, Germany. IUF-Leibniz Research Institute for Environmental Medicine, Duesseldorf, Germany.Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, P.O. Box 36, Byblos, Lebanon.Faculty of Health and Medical Sciences, Department of Microbial and Cellular Sciences, University of Surrey, Guildford, UK.Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, P.O. Box 36, Byblos, Lebanon.Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, P.O. Box 36, Byblos, Lebanon. cdaher@lau.edu.lb.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28073348

Citation

Shebaby, Wassim N., et al. "Wild Carrot Pentane-based Fractions Suppress Proliferation of Human HaCaT Keratinocytes and Protect Against Chemically-induced Skin Cancer." BMC Complementary and Alternative Medicine, vol. 17, no. 1, 2017, p. 36.
Shebaby WN, Mroueh MA, Boukamp P, et al. Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer. BMC Complement Altern Med. 2017;17(1):36.
Shebaby, W. N., Mroueh, M. A., Boukamp, P., Taleb, R. I., Bodman-Smith, K., El-Sibai, M., & Daher, C. F. (2017). Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer. BMC Complementary and Alternative Medicine, 17(1), 36. https://doi.org/10.1186/s12906-016-1531-0
Shebaby WN, et al. Wild Carrot Pentane-based Fractions Suppress Proliferation of Human HaCaT Keratinocytes and Protect Against Chemically-induced Skin Cancer. BMC Complement Altern Med. 2017 Jan 10;17(1):36. PubMed PMID: 28073348.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer. AU - Shebaby,Wassim N, AU - Mroueh,Mohamad A, AU - Boukamp,Petra, AU - Taleb,Robin I, AU - Bodman-Smith,Kikki, AU - El-Sibai,Mirvat, AU - Daher,Costantine F, Y1 - 2017/01/10/ PY - 2015/12/12/received PY - 2016/12/19/accepted PY - 2017/1/12/entrez PY - 2017/1/12/pubmed PY - 2017/1/21/medline KW - Daucus carota KW - HaCaT KW - TPA/DMBA skin cancer KW - Wild carrot SP - 36 EP - 36 JF - BMC complementary and alternative medicine JO - BMC Complement Altern Med VL - 17 IS - 1 N2 - BACKGROUND: Previous studies in our laboratory showed that the Lebanese Daucus carota ssp. carota (wild carrot) oil extract possesses in vitro and in vivo anticancer activities. The present study aims to examine the cytotoxic effect of Daucus carota oil fractions on human epidermal keratinocytes and evaluate the chemopreventive activity of the pentane diethyl ether fraction on DMBA/TPA induced skin carcinogenesis in mice. METHODS: Wild carrot oil extract was chromatographed to yield four fractions (F1, 100% pentane; F2, 50:50 pentane:diethyl ether; F3, 100% diethyl ether; F4 93:7 chloroform:methanol). The cytotoxic effect of fractions (10, 25, 50 and 100 μg/mL) was tested on human epidermal keratinocytes (non-tumorigenic HaCaT cells and tumorigenic HaCaT-ras variants) using WST a ssay. Cell cycle phase distribution of tumorigenic HaCaT-ras variants was determined by flow cytometry post-treatment with F2 fraction. Apoptosis related proteins were also assessed using western blot. The antitumor activity of F2 fraction was also evaluated using a DMBA/TPA induced skin carcinoma in Balb/c mice. RESULTS: All fractions exhibited significant cytotoxicity, with HaCaT cells being 2.4-3 times less sensitive than HaCaT-ras A5 (benign tumorigenic), and HaCaT-ras II4 (malignant) cells. GC-MS analysis revealed the presence of a major compound (around 60%) in the pentane/diethylether fraction (F2), identified as 2-himachalen-6-ol. Treatment of HaCaT-ras A5 and HaCaT-ras II4 cells with F2 fraction resulted in the accumulation of cells in the sub-G1 apoptotic phase and decreased the population of cells in the S and G2/M phases. Additionally, F2 fraction treatment caused an up-regulation of the expression of pro-apoptotic (Bax) and down-regulation of the expression of anti-apoptotic (Bcl2) proteins. A decrease in the phosphorylation of AKT and ERK was also observed. Intraperitoneal treatment with F2 fraction (50 or 200 mg/kg) in the DMBA/TPA skin carcinogenesis mouse model showed a significant inhibition of papilloma incidence (mice with papilloma), yield (number of papilloma/mouse) and volume (tumor relative size) at weeks 15, 18 and 21. CONCLUSION: The present data reveal that F2 fraction has a remarkable antitumor activity against DMBA/TPA-induced skin carcinogenesis, an effect that may be mediated through inhibition of the MAPK/ERK and PI3K/AKT pathways. SN - 1472-6882 UR - https://www.unboundmedicine.com/medline/citation/28073348/Wild_carrot_pentane_based_fractions_suppress_proliferation_of_human_HaCaT_keratinocytes_and_protect_against_chemically_induced_skin_cancer_ L2 - https://bmccomplementalternmed.biomedcentral.com/articles/10.1186/s12906-016-1531-0 DB - PRIME DP - Unbound Medicine ER -