Analysis of Kras gene from induced pancreatic cancer rats administered with Momordicacharantia and Ocimumbasilicum leaf extracts

10.1016/j.jtcme.2017.04.003

J.B. Minari；C.A. Okelola；N.C. Ugochukwu

K-Ras ； Momordica charantia ； Ocimum basilicum ； Pancreatic cancer ； Rats

Journal of Traditional and Complementary Medicine

8卷2期（2018 / 04 / 01）

282 - 288

英文

Objective: To analyze K-ras gene from induced pancreatic cancer rats administered with Momordicacharantia and Ocimumbasilicum leaf extracts. Methods: Twenty-five (25) adult rats weighing between 90-120 g were divided into 5 groups namely RA, RB, RC, NC and PC, each group had 5 rats. The PC which served as the control was fed with normal fish meal and water ad libitum; the NC which is the negative control received 20 mg/ml/week of Nitrosamines only while other groups received different concentrations of aqueous extract of both M. charantia and O. basilicum (200 mg, 100 mg, 50 mg) and Nitrosamine. Qualitative phytochemical screening of the aqueous extract of both M. charantia and O. basilicum was carried out. The extraction of DNA was done using Jena Bioscience DNA preparation kit and the protocol was based on the spin column based genomic DNA purification from blood, animal and plant cells. Agarose gel electrophoresis was used to analyze the K-ras gene extracted from the pancreas tissues of experimental rats while hematoxylinand eosin staining was used for histological assay. Results: Phytochemical screening revealed the presence of alkaloids, tannins, flavonoids, saponins and glycosides in M. charantia while saponins, tannins and glycosides were discovered in O. basilicum. Significant reduction in the weight of rats treated with 200 mg of aqueous extracts of M. charantia and O. basilicum while rats that were dosed with nitrosamines only showed a slight increase in weight in the first three weeks when compared to the positive control. Histological studies revealed that there is both enlargement and reduction in the islet cell size, with one of the sections showing a normal islet cell size. While the agarose gel electrophoresis revealed that there may be possibility of prevention of damage to k-ras gene as a result of the effect of plants extract. Conclusion: This work has shown that the leaf extracts of both M. charantia and O. basilicum will serve as a measure against induced pancreatic cancer in rats.

1. Alawode, TT(2013).An overview of the anti-cancer properties of some plants used in traditional medicine in Nigeria.Int Res J Biochem Bioinform,3(1),7-14.
2. Almeida, I,Alviano, DS,Vieira, DP(2007).Antigiardial activity of Ocimum basilicum essential oil.Parasitol Res,101(2),443-452.
3. American Cancer Society(2015).Cancer Facts & Figures.Atlanta, Ga:American Cancer Society.
4. Arthur, FKN,Woode, E,Terlabi, EO,Larbie, C(2011).Evaluation of acute and subchlornic toxicity of Annona muricata (Linn.) aqueous extract in animals.Eur J Exp Biol.,1(4),115-124.
5. Ayeni, MJ,Oyeyemi, SD,Kayode, J,Peter, GP(2015).Phytochemical, proximate and mineral anayses of the leaves of Gossypium hirsutum L. and Momordica charantia L.J Nat Sci Res,5(6),12-20.
6. Baines, AT,Xu, D,Der, CJ(2011).Inhibition of Ras for cancer treatment: the search continues.Future Med Chem,3,1787-1808.
7. Bakare, RI,Magbagbeola, OA,Akinwande, AI,Okunowo, OW(2010).Nutritional and chemical evaluation of Momordica charantia.J Med Plants Res,4(21),2189-2193.
8. Bozin, B,Mimica-Dukic, N,Simin, N,Anackov, G(2006).Characterization of the volatilecomposition of essential oils of some lamiaceae spices and the antimicrobial and antioxidant activities of the entire oils.J Agric Food Chem,54(5),1822-1828.
9. Chiang, LC,Ng, LT,Cheng, PW,Chiang, W,Lin, CC(2005).Antiviral activities of extracts and selected pure constituents of Ocimum basilicum.Clin Exp Pharmacol Physiol,32(10),811-816.
10. Daniel, VN,Daniang, IE,Nimyel, ND(2011).Phytochemical analysis and mineral elements composition of Ocimum basilicum obtained in JOS metropolis, Plateau State, Nigeria.Int J Eng Technol,11(6),135-137.
11. Duncan, DB(1955).Multiple range and multiple F test.Biometrics,11(1),1-42.
12. Hayashi, N,Egami, H,Ogawa, M(2002).Genetics of pancreatic cancer: recent advances in molecular diagnosis.Nihon Geka Gakkai Zasshi,103(6),476-481.
13. Hecht, SS,Chen, CB,Ornaf, RM,Hoffmann, D,Tso, TC(1978).Chemical studies on tobacco smoke LVI. Tobacco specific nitrosamines: origins, carcinogenicity and metabolism.IARC Sci Publ.,19,395-413.
14. Hruban, RH,Petersen, GM,Ha, PK,Kern, SE(1998).Genetics of pancreatic cancer. From genes to families.Surg Oncol Clin N Am,7(1),1-23.
15. Jemal, A,Siegel, R,Xu, J,Ward, E(2010).Cancer statistics, 2010.CA Cancer J Clin,60,277-300.
16. Kabashima, H,Miura, N,Shimizu, M(2010).,未出版
17. Kuno, T,Tsukamoto, T,Hara, A,Tanaka, T(2012).Cancer chemoprevention through the induction of apoptosis by natural compounds.J Biophys Chem,3,156-173.
18. Lu, X,Hsieh, TC,Wu, JM(2004).Equiguard suppresses androgen dependent LNCaP prostate cancer cell proliferation by targeting cell cycle control via downregulation of the retinoblastoma protein Rb and induction of apoptosis via the release of cytochrome c.Int J Oncol.,25,1801-1807.
19. Makri, O,Kintzios, S(2007).Botany, cultivation, pharmaceutical properties, and biotechnology.J Herbs Spices Med Plants,13,123-150.
20. Malumbres, M,Barbacid, M(2003).RAS oncogenes: the first 30 years.Nat Rev Cancer,3,459-465.
21. Mann, E,Smith, MJ,Hellier, J,Balabanovic, JA,Hamed, H,Grunfeld, EA(2012).Cognitive behavioral treatment for women who have menopausal symptoms after breast cancer treatment (MENOS 1): a randomized controlled trial.Lancet Oncol,13,309-318.
22. Manosroi, J,Dhumtanom, P,Manosroi, A(2006).Anti-proliferative activity of essential oil extracted from Thai medicinal plants on KB and P388 cell lines.Cancer Lett,235(1),114-120.
23. Mao, JJ,Palmer, CS,Healy, KE,Desai, K,Amsterdam, J(2011).Complementary and alternative medicine use among cancer survivors: a population-based study.J Cancer Surviv,5,8-17.
24. Michaud, DS,Giovannucci, E,Willett, WC,Colditz, GA,Stampfer, MJ,Fuchs, CS(2001).Physical activity, obesity, height, and the risk of pancreatic cancer.JAMA,286,921-929.
25. Michaud, DS,Skinner, HG,Wu, K(2005).Dietary patterns and pancreatic cancer risk in men and women.J Natl Cancer Inst,97,518-524.
26. Muller, L,Kasper, P,Muller-Tegethoff, K,Petr, T(1994).The genooxic potential in vitro and in vivo of the allyl benzene etheric oils estragole, basil oil and transanethole.Mutant Res,325,129-136.
27. Nandini, G,Anil, KM,Sunil, M(2014).Essential Oils and their constituents as anticancer agents: a mechanistic view.Biomed Res Int,2014
28. Crosta Peter. Pancreatic Cancer: Causes, Symptoms and Treatments; 2015. Accessed 23rd March, 2016 http://www.medicalnewstoday.com/info/pancreatic-cancer/
29. Rodenhuis, S,Slebos, RJ,Boot, AJ(1988).Incidence and possible clinical significance of K-ras oncogene activation in adenocarcinoma of the human lung.Cancer Res,48,5738-5741.
30. Salek, C,Zavoral, M,Benesova-Minarikova, L(2004).Detection of k-ras mutation in pancreatic cancer samples collected by fine needle aspiration biopsy: an intermedeiate report on the first results and experience.Folia Gastroenterol Hepatol,2(4),150-155.
31. Sofowora, A.(1984).Medicinal Plants and Traditional Medicine in Africa.New York:John Wiley and Sons.
32. Stathis, A,Moore, MJ(2010).Advanced pancreatic carcinoma: current treatment and future challenges.Nat Rev Clin Oncol,7,163-172.
33. Upadhyaya, P,Lindgren, BR,Hecht, SS(2009).Comparative levels of O6-methylguanine, pyridyloxobutyl and pyridylhydroxybutyl-DNA adducts in lung and liver of rats treated chronically with the tobacco-specific carcinogen 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone.Drug Metab Dispos,37(6),1147-1151.
34. Vimala, JR,Leema, RA,Raja, S(2012).A study on the phytochemical analysis and corrosion inhibition on mild steel by Annona muricata L. leaves extract in 1 N hydrochloricacid.Der Chem Sin,3(3),582-588.
35. Watson, IR,Takahashi, K,Futreal, A,Chin, L(2013).Emerging patterns of somatic mutations in cancer.Nat Rev Genet,14(10),703-718.
36. Woo, HD,Kim, J(2013).Dietary flavonoid intake and risk of stomach and colorectal cancer.World J Gastroenterol,19(7),101-1019.
37. Ye, F,Xui, L,Yi, J,Zhang, W,Zhang, DY(2002).Anticancer activity of Scutellaria baicalensis and its potential mechanism.J Altern Complement Med,8,567-572.