省產石花菜萃取物對細胞生長抑制的探討

本研究之目的乃在探討省產石花對於細胞之生長是否具有抑制作用，並且觀察其生長抑制之作用是否具有選擇性，同時並探討其是否因誘導細胞程序化凋亡(apoptosis)之故。石花菜分別以PBS及methanol萃取之，並依照一般食用方式製備石花凍，經冷凍乾燥後溶解於DMSO中而後偵測不同之萃取物對Hepa1(Murine hepatoma cell)、HL-60 Human promyelocytic leukemia cell) 二株癌細胞以及正常NIH3T3 Murine embryo fibroblast cell) 細胞生長的影響。細胞數目使用Coulter counter計算之，細胞增殖測試則使用MTS assay kit偵測，並以DNA電泳以及Annexin V-FITC螢光染色法觀察石花菜萃取物是否可誘導細胞產生apoptosis的現象。結果顯示，石花菜之PBS- soluble extract對3種細胞株之生長及增殖都沒有抑制的效果( p＞0.05) ；但其methanol-soluble extract對Hepa1及NIH3T3細胞之生長及增殖有抑制的作用 (p＜0.05)；且DMSO-soluble extract可抑制3種細胞株之生長及增殖的作用 (p＜0.05)。螢光染色及DNA電泳分析結果顯示，Hepa1及NIH3T3細胞經methanol-soluble extract處理，3種細胞株經DMSO-soluble extract處理後，都有annexin V positive反應，細胞之DNA亦有斷片之產生，顯示石花菜萃取物會誘導細胞apoptosis的發生。總言之，石花菜之methanol- 及DMSO-soluble extract具有抑制細胞生長的果，且其生長抑制作用可能由於其可誘導細胞apoptosis的發生，但是此作用並不具有選擇性。

關鍵字

石花菜；生長抑制；細胞凋亡；螢光染色法； DNA斷片

並列摘要

The objective of this study was to investigate the effect of Gelidium amansii on the cell growth and the potential role of apoptosis on the growth inhibition was also explored. Two lines of cancer cells, murine hepatoma cell (Hepa1) and human promeyolitic leukemia cell (HL-60), and one normal cell lines, murine embryo fibroblast cell (NIH3T3) were used in this study. The cells were treated with PBS-, methanol-, or DMSO extracts of Gelidium amansii, and the cell growth and the cell proliferation were measured. The results indicate the PBS - soluble extract did not have the effects on the all lines of cells, whereas methanol-soluble extract had the inhibitory effect on the Hepa1 and NIH3T3 cells (p＜0.05), and DMSO- soluble extract had inhibitory effects on all lines cells (p＜0.05). The methanol-soluble extract treated Hepa1 and NIH3T3 cells, and all lines of cells treated with DMSO-soluble fraction of the Gelidium amansii showed annexin V positive response, and had fragmented DNA ladders on agarose gels indicating that the cells underwent apoptosis. Gelidium amansii possesse the antiproliferation effect of cultured cells and this may due to the induction of the apoptosis pathway, this inhibitory effect did not show cell specificity.

並列關鍵字

Gelidium amansii ； growth inhibition ； apoptosis ； DNA fragmentation ； Annexin V staining摘要

參考文獻

Berridge MV and Tan AS, 1993. Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch. Biochem. Biophys. 3, 207 - 215 Diamandopoulus GT, 1996. Cancer: An historical perspective. Anticancer Res. 16, 1595 — 1602. Evan G and Littlewood T, 1998. A matter of life and cell death. Science. 281, 1317-1322. Fahey JW, Zhang Y and Talalay P, 1997. Broccoli sprouts: An exceptionally rich source of inducers of enzyme that protect against chemical carcinogens. Proc. Nad. Acad. Sci. 94, 10367 - 10372 Furusawa E and Furusawa S, 1985. Anticancer activity of a natural product, viva-natural, extracted from Undaria pinnantifida on intraperitoneally implanted Lewis lung carcinoma. Oncology. 42, 364- 369 Fukushima S, Takada N, Hori T and Wanibuchi H, 1997. Cancer prevention by organosulfur compounds from garlic and onion. J. Cell. Biochem. —Supplement. 27, 200 - 205. Gerhauser C, You M, Lin J, Moriarty RM, Hawthorne M, Mehta RG, Moon RC and Pezzuto, JM, 1997. Cancer chemoprevention potential of sulforamate, a novel analogue of sulforaphane that induces phase 2 drug-metabolizing enzymes. Cancer Res. 57(2) : 272-278. Harada H, Noro T and Kamei Y, 1997. Selectivity antitumor activity in vitro from marine algae form Japan coasts. Biol. Pharm. Bull. 20(5), 541- 546. Haskell CM, 1990. Cancer treatment 3rd edition. pp9 — 43. Henshaw EC, 1995. Clinical Oncology: a multidisciplinary approach for physicians and students. 7th edition. 23-24. Johnson PH and Grossman LI, 1977. Electrophoresis of DNA in agarose gels. Optimizing separation of conformational isomers of double- and single-stranded DNAs. Biochem. 16, 4217- 4224.

Google Scholar

國際替代計量

省產石花菜萃取物對細胞生長抑制的探討

未授權

主題瀏覽