Thyme essential oil inhibits proliferation of DLD-1 colorectal cancer cells through antioxidant effect
Öz
Abstract
Aim. It is a very hot topic to investigate the effects and the mechanisms of action of biological active extracts such as thyme essential oil on various cancer cells. This paper examines the ability of thyme essential oil to inhibit the growth of DLD-1 colorectal cancer cells and if it is the case, determine the contribution of antioxidant effect to this ability. Methods. To do this, we treated human DLD-1 colorectal cancer cells with thyme essential oil and assessed its effects on cell proliferation by using real time cell analyzing system (Xcelligence System). Furthermore, we applied the same concentrations to the L929 fibroblast cells in order to determine if thyme essential oil has cytotoxicity on normal cells in the same concentrations it showed anticancer effects. Then, antioxidant ability of thyme essential oil was evaluated by using DPPH assay. Results. Thyme essential oil treated DLD-1 cells exhibited decreasing cell index values in a concentration-dependent manner. An inhibitor concentration 50 (IC50) value of 0.347 mg/mL was achieved. On the other hand, when thyme essential oil was tested on L929 fibroblast cells, it was observed that thyme essential oil did not show cytotoxic effect on L929 fibroblast cells. IC50 value was 22 mg/mL for this experiment. Besides, thyme essential oil showed a strong antioxidant effect on DPPH assay. Conclusion. When the antiproliferative and the antioxidant effects of thyme essential oil are considered, it can be concluded that thyme essential oil may be an alternative option in the treatment of colorectal cancer.
Keywords: Thyme, cytotoxicity, DLD-1, colorectal cancer, oxidative stress, Xcelligence system
Özet
Amaç. Kekik uçucu yağı gibi biyolojik aktif ekstraktların çeşitli kanser hücreleri üzerindeki etkilerini ve etki mekanizmalarını araştırma konusu oldukça önemli bir konudur. Bu çalışmada kekik uçucu yağının insan DLD-1 kolorektal kanser hücrelerinin büyümesini inhibe edip etmediği, eğer ediyorsa bu inhibisyona antioksidan etkisinin katkısını belirlemek amaçlanmıştır. Yöntem. İnsan DLD-1 kolorektal kanser hücrelerine kekik uçucu yağı uygulayarak, uçucu yağın hücre proliferasyonu üzerindeki etkilerini gerçek zamanlı hücre analiz sistemi (Xcelligence Sistemi) ile değerlendirdik. Ayrıca kekik uçucu yağının antikanser etkinlik gösterdiği konsantrasyonların normal hücrelerde sitotoksik olup olmadığını belirlemek amacıyla L929 fibroblast hücrelerine de aynı konsantrasyonları uyguladık. Daha sonra kekik uçucu yağının antioksidan etkinliğini DPPH yöntemini kullanarak değerlendirdik. Bulgular. Kekik uçucu yağı uygulanan DLD-1 hücreleri, konsantrasyona bağımlı olarak hücre indeksinde azalan değerler gösterdi. IC50 değeri 0,347 mg/mL olarak bulundu. Diğer taraftan kekik uçucu yağı L929 fibroblast hücreleri üzerine uygulandığında sitotoksik etkisi gözlenmedi. Bu deney için IC50 değeri 22 mg/mL olarak bulundu. Bunun yanında kekik uçucu yağı DPPH yönteminde güçlü antioksidan etkinlik gösterdi. Sonuç. Kekik uçucu yağının antiproliferatif ve antioksidan etkileri dikkate alındığında, kolorektal kanser tedavisinde alternatif bir seçenek olabileceği sonucuna varılmaktadır.
Anahtar sözcükler: Kekik, sitotoksisite, kolorektal kanser, oksidatif stress, Xcelligence system
Anahtar Kelimeler
Thyme cytotoxicity DLD-1 colorectal cancer oxidative stress Xcelligence system
Kaynakça
- Anand P, Kunnumakkara AB, Sundaram C, Harikumar KB, Tharakan ST, Lai OS, Sung B, Aggarwal BB. Cancer is a Preventable Disease that Requires Major Lifestyle Changes. Pharm Res 2008; 25: 2097-116.
- Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global Cancer Statistics. Ca Cancer J Clin 2011; 61: 69-90.
- Erarslan E, Yüksel İ, Haznedaroğlu S. Kolorektal karsinogenez ve metabolik sendrom ilişkisi. Cumhuriyet Med J 2012; 34: 380-5.
- Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007; 39: 44-84.
- Triantafillidis JK, Nasioulas G, Kosmidis PA. Colorectal Cancer and Inflammatory Bowel Disease: Epidemiology, Risk Factors, Mechanisms of Carcinogenesis and Prevention Strategies. Anticancer Res 2009; 29: 2727Ait M'barek L, Ait Mouse H, Jaâfari A, Aboufatima R, Benharref A, Kamal M, Bénard J, El Abbadi N, Bensalah M, Gamouh A, Chait A, Dalal A, Zyad A. Cytotoxic effect of essential oil of thyme (Thymus broussonettii) on the IGR-OV1 tumor cells resistant to chemotherapy. Braz J Med Biol Res 2007; 40: 1537-44.
- Badisa RB, Tzakou O, Couladis M, Pilarinou E. Cytotoxic Activities of some Greek Labiatae Herbs. Phytother Res 2003; 17: 472-6.
- Lee SJ, Umano K, Shibamoto T, Lee KG. Identification of volatile components in basil (Ocimum basilicum L.) and thyme leaves (Thymus vulgaris L.) and their antioxidant properties. Food Chemistry 2005; 91: 131Burits M, Bucar F. Antioxidant Activity of Nigella sativa Essential Oil. Phytother Res 2000; 14: 323-8.
- Roche Diagnostics GmbH. Introduction of the RTCA SP Instrument. RTCA SP Instrument Operator‘s Manual, A. Acea Biosciences, Inc 2008; 14SEER Stat Fact Sheet; colon and rectum. National Cancer Institute. (Accessed on February 20, 2013).
- O’Connell JB, Maggard MA, Ko CY. Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst 2004; 96: 1420-5.
- Kabbinavar FF, Hambleton J, Mass RD, Hurwitz HI, Bergsland E, Sarkar S.Combined analysis of efficacy: the addition of bevacizumab to fluorouracil/ leucovorin improves survival for patients with metastatic colorectal cancer. J Clin Oncol 2005; 23: 3706-12.
- Fuchs CS, Marshall J, Barrueco J. Randomized, controlled trial of irinotecan plus Infusional, bolus, or oral Fluoropyrimidines in first-line treatment of metastatic colorectal cancer: updated results from the BICC-C study. J Clin Oncol 2008; 26: 689-90.
- Hedberg I. Botanical methods in ethnopharmacology and the need for conservation of medicinal plants. J Ethnopharmacol 1993; 38: 121-8.
- Heinrich M, Gibbons S. Ethnopharmacology in drug discovery: an analysis of its role and potential contribution. J Pharm Pharmacol 2001; 53: 425-32.
- Ao Y, Satoh K, Shibano K, Kawahito Y, Shioda S. Singlet oxygen scavenging activity and cytotoxicity of essential oils from rutaceae. J Clin Biochem Nutr 2008; 43: 6-12.
- Pinn G. Herbs used in obstetrics and gynaecology. Aust Fam Physician 2001; 30: 351-4, 356.
- Ankli A, Heinrich M, Bork P, Wolfram L, Bauerfeind P, Brun R, Schmid C, Weiss C, Bruggisser R, Gertsch J, Wasescha M, Sticher O. Yucatec Mayan medicinal plants: evaluation based on indigenous uses. J Ethnopharmacol 2002; 79: 43-52.
- Manosroi J, Dhumtanom P, Manosroi A. Anti-proliferative activity of essential oil extracted from Thai medicinal plants on KB and P388 cell lines. Cancer Lett 2006; 235: 114-20.
- Yoo HH, Park JH, Kwon SW. In vitro cytotoxic activity of some Korean medicinal plants on human cancer cell lines: enhancement in cytotoxicity by heat processing. Phytother Res 2007; 21: 900-3.
- Aponte JC, Vaisberg AJ, Rojas R, Caviedes L, Lewis WH, Lamas G, Sarasara C, Gilman RH, Hammond GB. Isolation of cytotoxic metabolites from targeted peruvian amazonian medicinal plants. J Nat Prod 2008; 71: 102Lampronti I, Saab AM, Gambari R. Antiproliferative activity of essential oils derived from plants belonging to the Magnoliophyta division. Int J Oncol 2006; 29: 989-95.
- Albuquerque MR, Costa SM, Bandeira PN, Santiago GM, Andrade-Neto M, Silveira ER, Pessoa OD. Nematicidal and larvicidal activities of the essential oils from aerial parts of Pectis oligocephala and Pectis apodocephala Baker. An Acad Bras Cienc 2007; 79: 209-13.
- Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils--a review. Food Chem Toxicol 2008; 46: 446-75.
- Baik JS, Kim SS, Lee JA, Oh TH, Kim JY, Lee NH, Hyun CG. Chemical composition and biological activities of essential oils extracted from Korean endemic citrus species. J Microbiol Biotechnol 2008; 18: 74-9.
- Berrington D, Lall N. Anticancer Activity of Certain Herbs and Spices on the Cervical Epithelial Carcinoma (HeLa) Cell Line. Evid Based Complement Alternat Med 2012; 564927.
- Sertel S, Eichhorn T, Plinkert PK, Efferth T. Cytotoxicity of Thymus vulgaris essential oil towards human oral cavity squamous cell carcinoma. Anticancer Res 2011; 31: 1: 81-7.
- Yi W, Wetzstein HY. Anti-tumorigenic activity of five culinary and medicinal herbs grown under greenhouse conditions and their combination effects. J Sci Food Agric 2011; 15: 91: 1849-54.
- Roessner A, Kuester D, Malfertheiner P, Schneider-Stock R. Oxidative stress in ulcerative colitis-associated carcinogenesis. Pathol Res Pract 2008; 204: 511-24.
- Kraus S, Arber N. Inflammation and colorectal cancer. Curr Opin Pharmacol 2009; 9: 405-10.
- Seril DN, Liao J, Yang GY. Colorectal carcinoma development in inducible nitric oxide synthase-deficient mice with dextran sulfate sodiuminduced ulcerative colitis. Mol Carcinog 2007; 46: 341-53.
- El-Nekeety AA, Mohamed SR, Hathout AS, Hassan NS, Aly SE, AbdelWahhab MA. Antioxidant properties of Thymus vulgaris oil against aflatoxin-induce oxidative stress in male rats. Toxicon 2011; 57: 984-91. Undeğer U, Başaran A, Degen GH, Başaran N. Antioxidant activities of major thyme ingredients and lack of (oxidative) DNA damage in V79 Chinese hamster lung fibroblast cells at low levels of carvacrol and thymol. Food Chem Toxicol 2009; 47:2037-43
Öz
Amaç. Kekik uçucu yağı gibi biyolojik aktif ekstraktların çeşitli kanser hücreleri üzerindeki etkilerini ve etki mekanizmalarını araştırma konusu oldukça önemli bir konudur. Bu çalışmada kekik uçucu yağının insan DLD-1 kolorektal kanser hücrelerinin büyümesini inhibe edip etmediği, eğer ediyorsa bu inhibisyona antioksidan etkisinin katkısını belirlemek amaçlanmıştır. Yöntem. İnsan DLD-1 kolorektal kanser hücrelerine kekik uçucu yağı uygulayarak, uçucu yağın hücre proliferasyonu üzerindeki etkilerini gerçek zamanlı hücre analiz sistemi (Xcelligence Sistemi) ile değerlendirdik. Ayrıca kekik uçucu yağının antikanser etkinlik gösterdiği konsantrasyonların normal hücrelerde sitotoksik olup olmadığını belirlemek amacıyla L929 fibroblast hücrelerine de aynı konsantrasyonları uyguladık. Daha sonra kekik uçucu yağının antioksidan etkinliğini DPPH yöntemini kullanarak değerlendirdik. Bulgular. Kekik uçucu yağı uygulanan DLD-1 hücreleri, konsantrasyona bağımlı olarak hücre indeksinde azalan değerler gösterdi. IC değeri 0,347 mg/mL olarak bulundu. Diğer taraftan kekik uçucu yağı L929 fibroblast hücreleri üzerine uygulandığında sitotoksik etkisi gözlenmedi. Bu deney için IC değeri 22 mg/mL olarak bulundu. Bunun yanında kekik uçucu yağı DPPH yönteminde güçlü antioksidan etkinlik gösterdi. Sonuç. Kekik uçucu yağının antiproliferatif ve antioksidan etkileri dikkate alındığında, kolorektal kanser tedavisinde alternatif bir seçenek olabileceği sonucuna varılmaktadır.
Anahtar Kelimeler
Kekik sitotoksisite kolorektal kanser oksidatif stress Xcelligence system
Kaynakça
- Anand P, Kunnumakkara AB, Sundaram C, Harikumar KB, Tharakan ST, Lai OS, Sung B, Aggarwal BB. Cancer is a Preventable Disease that Requires Major Lifestyle Changes. Pharm Res 2008; 25: 2097-116.
- Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global Cancer Statistics. Ca Cancer J Clin 2011; 61: 69-90.
- Erarslan E, Yüksel İ, Haznedaroğlu S. Kolorektal karsinogenez ve metabolik sendrom ilişkisi. Cumhuriyet Med J 2012; 34: 380-5.
- Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007; 39: 44-84.
- Triantafillidis JK, Nasioulas G, Kosmidis PA. Colorectal Cancer and Inflammatory Bowel Disease: Epidemiology, Risk Factors, Mechanisms of Carcinogenesis and Prevention Strategies. Anticancer Res 2009; 29: 2727Ait M'barek L, Ait Mouse H, Jaâfari A, Aboufatima R, Benharref A, Kamal M, Bénard J, El Abbadi N, Bensalah M, Gamouh A, Chait A, Dalal A, Zyad A. Cytotoxic effect of essential oil of thyme (Thymus broussonettii) on the IGR-OV1 tumor cells resistant to chemotherapy. Braz J Med Biol Res 2007; 40: 1537-44.
- Badisa RB, Tzakou O, Couladis M, Pilarinou E. Cytotoxic Activities of some Greek Labiatae Herbs. Phytother Res 2003; 17: 472-6.
- Lee SJ, Umano K, Shibamoto T, Lee KG. Identification of volatile components in basil (Ocimum basilicum L.) and thyme leaves (Thymus vulgaris L.) and their antioxidant properties. Food Chemistry 2005; 91: 131Burits M, Bucar F. Antioxidant Activity of Nigella sativa Essential Oil. Phytother Res 2000; 14: 323-8.
- Roche Diagnostics GmbH. Introduction of the RTCA SP Instrument. RTCA SP Instrument Operator‘s Manual, A. Acea Biosciences, Inc 2008; 14SEER Stat Fact Sheet; colon and rectum. National Cancer Institute. (Accessed on February 20, 2013).
- O’Connell JB, Maggard MA, Ko CY. Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst 2004; 96: 1420-5.
- Kabbinavar FF, Hambleton J, Mass RD, Hurwitz HI, Bergsland E, Sarkar S.Combined analysis of efficacy: the addition of bevacizumab to fluorouracil/ leucovorin improves survival for patients with metastatic colorectal cancer. J Clin Oncol 2005; 23: 3706-12.
- Fuchs CS, Marshall J, Barrueco J. Randomized, controlled trial of irinotecan plus Infusional, bolus, or oral Fluoropyrimidines in first-line treatment of metastatic colorectal cancer: updated results from the BICC-C study. J Clin Oncol 2008; 26: 689-90.
- Hedberg I. Botanical methods in ethnopharmacology and the need for conservation of medicinal plants. J Ethnopharmacol 1993; 38: 121-8.
- Heinrich M, Gibbons S. Ethnopharmacology in drug discovery: an analysis of its role and potential contribution. J Pharm Pharmacol 2001; 53: 425-32.
- Ao Y, Satoh K, Shibano K, Kawahito Y, Shioda S. Singlet oxygen scavenging activity and cytotoxicity of essential oils from rutaceae. J Clin Biochem Nutr 2008; 43: 6-12.
- Pinn G. Herbs used in obstetrics and gynaecology. Aust Fam Physician 2001; 30: 351-4, 356.
- Ankli A, Heinrich M, Bork P, Wolfram L, Bauerfeind P, Brun R, Schmid C, Weiss C, Bruggisser R, Gertsch J, Wasescha M, Sticher O. Yucatec Mayan medicinal plants: evaluation based on indigenous uses. J Ethnopharmacol 2002; 79: 43-52.
- Manosroi J, Dhumtanom P, Manosroi A. Anti-proliferative activity of essential oil extracted from Thai medicinal plants on KB and P388 cell lines. Cancer Lett 2006; 235: 114-20.
- Yoo HH, Park JH, Kwon SW. In vitro cytotoxic activity of some Korean medicinal plants on human cancer cell lines: enhancement in cytotoxicity by heat processing. Phytother Res 2007; 21: 900-3.
- Aponte JC, Vaisberg AJ, Rojas R, Caviedes L, Lewis WH, Lamas G, Sarasara C, Gilman RH, Hammond GB. Isolation of cytotoxic metabolites from targeted peruvian amazonian medicinal plants. J Nat Prod 2008; 71: 102Lampronti I, Saab AM, Gambari R. Antiproliferative activity of essential oils derived from plants belonging to the Magnoliophyta division. Int J Oncol 2006; 29: 989-95.
- Albuquerque MR, Costa SM, Bandeira PN, Santiago GM, Andrade-Neto M, Silveira ER, Pessoa OD. Nematicidal and larvicidal activities of the essential oils from aerial parts of Pectis oligocephala and Pectis apodocephala Baker. An Acad Bras Cienc 2007; 79: 209-13.
- Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils--a review. Food Chem Toxicol 2008; 46: 446-75.
- Baik JS, Kim SS, Lee JA, Oh TH, Kim JY, Lee NH, Hyun CG. Chemical composition and biological activities of essential oils extracted from Korean endemic citrus species. J Microbiol Biotechnol 2008; 18: 74-9.
- Berrington D, Lall N. Anticancer Activity of Certain Herbs and Spices on the Cervical Epithelial Carcinoma (HeLa) Cell Line. Evid Based Complement Alternat Med 2012; 564927.
- Sertel S, Eichhorn T, Plinkert PK, Efferth T. Cytotoxicity of Thymus vulgaris essential oil towards human oral cavity squamous cell carcinoma. Anticancer Res 2011; 31: 1: 81-7.
- Yi W, Wetzstein HY. Anti-tumorigenic activity of five culinary and medicinal herbs grown under greenhouse conditions and their combination effects. J Sci Food Agric 2011; 15: 91: 1849-54.
- Roessner A, Kuester D, Malfertheiner P, Schneider-Stock R. Oxidative stress in ulcerative colitis-associated carcinogenesis. Pathol Res Pract 2008; 204: 511-24.
- Kraus S, Arber N. Inflammation and colorectal cancer. Curr Opin Pharmacol 2009; 9: 405-10.
- Seril DN, Liao J, Yang GY. Colorectal carcinoma development in inducible nitric oxide synthase-deficient mice with dextran sulfate sodiuminduced ulcerative colitis. Mol Carcinog 2007; 46: 341-53.
- El-Nekeety AA, Mohamed SR, Hathout AS, Hassan NS, Aly SE, AbdelWahhab MA. Antioxidant properties of Thymus vulgaris oil against aflatoxin-induce oxidative stress in male rats. Toxicon 2011; 57: 984-91. Undeğer U, Başaran A, Degen GH, Başaran N. Antioxidant activities of major thyme ingredients and lack of (oxidative) DNA damage in V79 Chinese hamster lung fibroblast cells at low levels of carvacrol and thymol. Food Chem Toxicol 2009; 47:2037-43
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Bölüm | Temel Tıp Bilimleri Araştırma Yazıları |
| Yazarlar | |
| Yayımlanma Tarihi | 22 Mart 2013 |
| Yayımlandığı Sayı | Yıl 2013Cilt: 35 Sayı: 1 |