數株乳酸桿菌細胞區分物抑制大腸癌細胞增生及抗細胞毒性研究

Study on the Antiproliferation of Colon Cancer Cells and Anticytotoxicity of Various Cell Fractions from Lactobacillus spp.

林佩儀(Pei-Yi Lin)

指導教授 ： 游若篍

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.10006

摘要

大腸結腸癌為台灣惡性腫瘤十大死因之一，相關研究指出，乳酸菌可以和致突變物質進行結合的作用，降低致突變物質對腸道細胞組織受到致癌物的傷害，具有抗氧化活性、抗致突變性和抗腫瘤等功能。因此本研究利用三株乳酸菌Lactobacillus casei 01、L. plantarum、L. acidophillus為材料，進行細胞試驗，結果顯示乳酸菌之不同細胞區分物不會對人類腸道上皮細胞 Int-407 生長造成影響。抑制癌細胞增生的實驗結果，乳酸菌之不同區分物經過48小時作用之後，可顯著的下降大腸癌細胞HT-29 及 Caco-2之存活率，最低達 33.0% 與54.8%，為 L. casei 01 與 L. acidophilus之熱致死細胞作用結果。此外，利用彗星電泳法 (Comet assay) 評估樣品降低化學致突變劑 4NQO 誘導 Int-407產生細胞損傷及基因毒性之效果。進一步利用分段式預反應，探討其抗細胞毒性機制。最後以化學抗氧化方法檢測包括螯合亞鐵離子能力以及DPPH自由基捕捉能力。三株乳酸菌之胞內萃取液皆可顯著降低4NQO誘導人體腸道細胞Int-407株之DNA損傷，細胞壁部分則是 L. casei 01，損傷分數由172.2下降至71.7最為顯著。並發現乳酸菌胞內液其抗細胞毒性之機制主要為生物抗細胞毒性(Bioanticytotoxic effect) 以及去細胞毒性 (Descytotoxic) 作用。化學抗氧化試驗中，與亞鐵離子螯合能力最佳的是 L. plantarum，其熱致死細胞、粗細胞壁與之螯合力分別為 51.8、55.6 及 56.8%。對DPPH之清除能力，當三株乳酸菌之菌數為109 CFU/mL時，其熱致死細胞部位對於清除 DPPH 自由基之能力達33.7%-46.4%。

關鍵字

Lactobacillus ； 彗星電泳 ； 抗細胞毒性 ； 抗癌細胞增生

並列摘要

Colon cancer is a major cause of death in Taiwan. Recently, studies on lactic acid bacteria have indicated that they possess antioxidative, antimutagenic, and antitumor activities. In this study, the cell fractions (heat-killed cells, crude cell walls and intracellular extracts) of Lactobacillus acidophilus, L. casei 01, and L. plantarum were studied for their effects on the growth of human intestinal epithelial cells, intestine 407 and antiproliferation of human colon cancer cells, HT-29 and Caco-2. The viability of intestine 407 cells was not affected by all cell fractions (p<0.05). And the results showed that, after treated with 48h, all the cell fractions of three Lactobacillus spp. could reduce the cell viability in HT-29 and Caco-2 cells(p<0.05). The lowest in HT-29 and Caco-2 cells are 33.0% and 54.8%, after treated with the heat-killed cells of L. casei 01 and L. acidophilus BCRC 14079. Furthermore, the prevention of 4NQO-induced DNA damage on intestine 407 cells was studied as by comet assay. The possible anticytotoxic mechanisms of cell fractions were also investigated by section-preincubation test. The intracellular extract of three Lactobacillus spp. decreased the 4NQO-induced DNA damage on Int-407 (p<0.05). As in the part of crude cell wall, L. casei 01 has the most significantly effect, reduce the DNA damage score from 172.2 to 78.1. And the primary mechanism of intracellular extract of three Lactobacillus spp. were bioanticytotoxic effect and descytotoxic effect. In addition, all of the three Lactobacillus spp. have antioxidative abilities, the cell fractions of L. plantarum, has the chelating effects of ferrous ions between 51.8-56.8%, was the most significantly one. The scavenging effects of 2-Dipheny-1-picryl-hydrazyl (DPPH), showed the heat-killed cells of 109 CFU/mL Lactobacillus spp. were the most effective one, scavenged from 33.7-46.4%.

並列關鍵字

Lactobacillus ； Comet assay ； anticytotoxic ； antiproliferation

參考文獻

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