BMC Complementary and Alternative Medicine | |
Cephalotaxus griffithii Hook.f. needle extract induces cell cycle arrest, apoptosis and suppression of hTERT and hTR expression on human breast cancer cells | |
Narayan Chandra Talukdar2  Mohan Chandra Kalita1  Jagat Chandra Borah2  Surbala Laishram2  Dinesh Singh Moirangthem2  | |
[1] Department of Biotechnology, Gauhati University, Guwahati 781014, Assam, India;Institute of Bioresources and Sustainable Development, Takyelpat Institutional Area, Imphal 795001, Manipur, India | |
关键词: p53; Telomerase; Cell cycle; Apoptosis; Cephalotaxus griffithii; | |
Others : 1086940 DOI : 10.1186/1472-6882-14-305 |
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received in 2014-04-30, accepted in 2014-08-13, 发布年份 2014 | |
【 摘 要 】
Background
Cephalotaxus spp. are known to possess anticancer potential. In this present work, for the first time the effects of C. griffithii needle (CGN) extracts on human cancer cells were examined.
Methods
The CGN was successively extracted with petroleum ether (PE), acetone and methanol. The extracts were tested for its effect on proliferation of cancer cells (MTT assay on HeLa, ZR751 and HepG2). Extract that showed the maximum growth inhibitory effect was subjected for mechanism of action study. These included apoptosis (morphological and DNA fragmentation assay), cell cycle (flow cytometry), caspase expression (Western blot) and activity (assay kit), p53 (western blot and TP53 siRNA interference) and telomerase expression (reverse transcriptase PCR) analysis.
Results
Among the extracts, PE extract induced maximum cytotoxicity, with highest death occurred in ZR751 cells. Since, PE extract induced cell death was highest among the CGN extracts, with maximum cancer cell death occurred in ZR751 cells; we carried out mechanism study of PE extract induced ZR751 cell death. It was observed that PE extract induced ZR751 cell death was associated with cell cycle arrest and apoptosis by activating both intrinsic and extrinsic apoptotic pathways. Knock down study revealed that p53 is essential for loss of ZR751 cell viability induced by PE extract. Further, PE extract down-regulated hTERT, hTR, and c-Myc expression. Thin layer chromatography analysis indicated the presence of unique phytochemicals in PE extract.
Conclusion
Based on the observations, we concluded that PE extract of C. griffithii needle contains important phyto-components with multiple cellular targets for control of breast cancer and is worthy of future studies.
【 授权许可】
2014 Moirangthem et al.; licensee BioMed Central Ltd.
【 预 览 】
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【 参考文献 】
- [1]American Cancer Society: Global Cancer Facts and Figures 2007. Atlanta: American Cancer Society; 2007.
- [2]Newman DJ, Cragg GM: Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod 2003, 75:311-335.
- [3]Takano I, Yasuda I, Nishijima M, Hitotsuyanagi Y, Takeya K, Itokawa H: Drupangtonine, a novel antileukemic alkaloid from Cephalotaxus harringtonia var. drupacea. Bioorg Med Chem Lett 1996, 6:1689-1690.
- [4]Takano I, Yasuda I, Nishijima M, Yanagi Y, Takeya K, Itokawa H: Ester-type Cephalotaxus alkaloids from Cephalotaxus harringtonia var. Drupacea. Phytochemistry 1997, 44:735-738.
- [5]Kuo YH, Lin CH, Hwang SY, Shen YC, Lee YL, Shyh-Yuan L: A novel cytotoxic C-methylated biflavone from the stem of Cephalotaxus wilsoniana. Chem Pharm Bull 2000, 48:440-441.
- [6]Morita H, Yoshinaga M, Kobayashi J: Cephalezomines G, H, J, K, L. and M, new alkaloids from Cephalotaxus harringtonia var. nana. Tetrahedron 2002, 58:5489-5495.
- [7]U.S. Food and Drug Administration: FDA approves Synribo for chronic myelogenous leukemia. 2012. Available: http://www.fda.gov/NewsEvents/%20Newsroom/%20PressAnnouncements/ucm325895.htm webcite. Accessed 14 January 2014
- [8]Uma S: Blooming potential of the detached shoots of Cephalotaxus griffithii Hook. f. Curr Sci 2008, 95:320-321.
- [9]Kamil M, Khan NA, Ahmad I, Iiyas M, Rahman W: Chemical constituents of Cephalotaxus griffithii. J Indian Chem Soc 1982, 59:1199.
- [10]Phutdhawong W, Korth J, Budhasukh D, Pyne SG: Volatile components from Cephalotaxus griffithii growing in northern Thailand. Flavour Fragr J 2002, 179:153-155.
- [11]Moirangthem DS, Talukdar NC, Kasoju N, Bora U: Antioxidant, antibacterial, cytotoxic, and apoptotic activity of stem bark extracts of Cephalotaxus griffithii Hook. F. BMC Complement Altern Med 2012, 12:30. doi:10.1186/1472-6882-12-30 BioMed Central Full Text
- [12]Kayani SA, Masood A, Achakzai AKK, Anbreen S: Distribution of secondary metabolites in plants of quetta-balochistan. Pak J Bot 2007, 39:1173-1179.
- [13]Achakzai AKK, Achakzai P, Masood A, Kayani SA, Tareen RB: Response of plant parts and age on the distribution of secondary metabolites on plants found in quetta. Pak J Bot 2009, 41:2129-2135.
- [14]Hyder PW, Fredrickson EL, Estell RE, Tellez M, Gibben RP: Distribution and concentration of total phenolics, condensed tannins, and nordihydroguaiaretic acid (NDGA) in creosotebush (Larrea tridentate). Biochem Sys Ecol 2002, 30:905-912.
- [15]Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65:55-63.
- [16]Ribble D, Goldstein NB, Norris DA, Shellman YG: A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnol 2005, 5:12. doi:10.1186/1472-6750-5-12 BioMed Central Full Text
- [17]Riccardi C, Nicoletti I: Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 2006, 1:1458-1461.
- [18]Fido J, Tatham S, Shewry R: Western blotting analysis. Methods Mol Biol 1995, 49:423-437.
- [19]Fuke H, Shiraki K, Sugimoto K, Tanaka J, Beppu T, Yoneda K, Yamamoto N, Ito K, Masuya M, Takei Y: Jak inhibitor induces S phase cell-cycle arrest and augments TRAIL-induced apoptosis in human hepatocellular carcinoma cells. Biochem Biophys Res Commun 2007, 363:738-744.
- [20]Chang JS, Son JK, Li G, Oh EJ, Kim JY, Park SH, Bae JT, Kim HJ, Lee IS, Kim OM, Kozukue N, Han JS, Hirose M, Lee KR: Inhibition of cell cycle progression on HepG2 cells by hypsiziprenol A9, isolated from Hypsizigus marmoreus. Cancer Lett 2004, 212:7-14.
- [21]Ye M, Liu JK, Lu ZX, Zhao Y, Liu SF, Li LL, Tan M, Weng XX, Li W, Cao Y: Grifolin, a potential antitumor natural product from the mushroom Albatrellus confluens, inhibits tumor cell growth by inducing apoptosis in vitro. FEBS Lett 2005, 579:3437-3443.
- [22]Hsieh TC, Wu P, Park S, Wu JM: Induction of cell cycle changes and modulation of apoptogenic/anti-apoptotic and extracellular signaling regulatory protein expression by water extracts of I’m-YunityTM (PSP). BMC Complement Altern Med 2006, 6:30. doi:10.1186/1472-6882-6-30 BioMed Central Full Text
- [23]Bortner CD, Oldenburg NB, Cidlowski JA: The role of DNA fragmentation in apoptosis. Trends Cell Biol 1995, 5:21-26.
- [24]Yu J, Liu H, Lei J, Tan W, Hu X, Zou G: Antitumor activity of chloroform fraction of Scutellaria barbata and its active constituents. Phytother Res 2007, 21:817-822.
- [25]Taylor WR, Stark GR: Regulation of the G2/M transition by p53. Oncogene 2001, 20:1803-1815.
- [26]Ghobrial IM, Witzig TE, Adjei AA: Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin 2005, 55:178-194.
- [27]Bartek J, Iggo R, Gannon J, Lane DP: Genetic and immunochemical analysis of mutant p53 in human breast cancer cell lines. Oncogene 1990, 5:893-909.
- [28]Yu J, Zhang L, Hwang PM, Kinzler KW, Vogelstein B: PUMA induces the rapid apoptosis of colorectal cancer cells. Mol Cell 2001, 7:673-682.
- [29]Makin G, Dive C: Apoptosis and cancer chemotherapy. Trends Cell Biol 2001, 11:S22-S26.
- [30]Murray-Zmijewski F, Slee EA, Lu X: A complex barcode underlies the heterogeneous response of p53 to stress. Nat Rev Mol Cell Biol 2008, 9:702-712.
- [31]Scovassi AI, Poirier GG: Poly (ADP-ribosylation) and apoptosis. Mol Cell Biochem 1999, 199:125-127.
- [32]Mandal M, Kumar R: Bcl2 modulates telomerase activity. J Biol Chem 1997, 72:14183-14187.
- [33]Hahn WC, Stewart SA, Brooks MW, York SG, Eaton E, Kurachi A, Beijersbergen RL, Knoll JH, Meyerson M, Weinberg RA: Inhibition of telomerase limits the growth of human cancer cells. Nat Med 1999, 5:1164-1170.
- [34]Saretzki G: Telomerase, mitochondria and oxidative stress. Exp Gerontol 2009, 44:485-492.
- [35]Hahn WC, Meyerson M: Telomerase activation, cellular immortalization and cancer. Ann Med 2001, 33:123-129.
- [36]Oulton R, Harrington L: Telomeres, telomerase, and cancer: life on the edge of genomic stability. Curr Opin Oncol 2000, 12:74-81.
- [37]Kirkpatrick KL, Clark G, Ghilchick M, Newbold RF, Mokbel K: hTERT mRNA expression correlates with telomerase activity in human breast cancer. Eur J Surg Oncol 2003, 29:321-326.
- [38]Casillas MA, Brotherton SL, Andrews LG, Ruppert JM, Tollefsbol TO: Induction of endogenous telomerase (hTERT) by c-Myc in WI-38 fibroblasts transformed with specific genetic elements. Gene 2003, 316:57-65.