3-黃酮醇抑制人類骨肉瘤細胞143B之侵襲及活體腫瘤之生長能力

3-hydroxyflavone Reduces Cell Invasion in Human Osteosarcoma 143B Cells and Inhibit Tumor Growth In Vivo

鄭富元(Fu-Yuan Cheng)

指導教授 ： 謝易修

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2015年)

https://doi.org/10.6834/CSMU.2015.00106

摘要

骨肉瘤Osteosarcoma (OS)，是原發性骨瘤中最常見，惡性程度最高的骨腫瘤，常見於膝關節，這是因為膝關節附近的生長板成長得最快，且病情進展迅速容易發生肺轉移及肝轉移，是導致病人死亡的主要原因。類黃酮之 C-3 位置上有羥基 (hydroxy；OH )者稱之為黃酮醇，3-hydroxyflavone為黃酮醇的結構骨架，已有報告指出可抑制乳癌、前列腺癌細胞和大腸癌細胞的增生，對於人類骨肉瘤細胞亦有所抑制作用。本實驗研究3-hydroxyflavone是否能抑制人類骨肉瘤細胞143B的侵襲與轉移，以免疫缺陷小鼠(BALB/cAnN.CgFoxnlnu/CrlNarl)進行動物實驗，觀察注射癌細胞後，老鼠體重變化、體表腫瘤生長、腫瘤組織大小，再利用活體影像分析系統 (IVIS)進一步觀察腫瘤在活體內的生長狀況及癌轉移，比較管餵3-hydroxyflavone的實驗組與未餵食的控制組，結果發現3-hydroxyflavone具有抑制骨肉瘤大小，但是針對癌轉移觀察並沒有產生癌細胞的轉移現象。再利用143B細胞探討 3-hydroxyflavone是否會抑制細胞的侵襲及轉移，分別以MTT assays、modified Boyden chamber assay 、zymography assay及細胞傷口癒合分析，發現3-hydroxyflavone無細胞毒性，可以降低骨癌細胞中MMP-2的表現，對細胞的移動與侵襲能力具有抑制的作用。由以上實驗結果推測在人類骨肉瘤細胞的侵襲能力上，3-hydroxyflavone可能具備抑制效果，但仍需進一步進行長期動物實驗追蹤與蛋白分析，取得更有利證據以期應用到臨床治療。

關鍵字

骨肉瘤 ； 類黃酮 ； 黃酮醇

並列摘要

Osteosarcoma (OS) is the most common primary osteoma with the highest level of malignancy. It frequently occurs at knees because knees contain growth plates with the fastest growing rate. The disease progresses rapidly and which make lung and liver metastases occur easily, which are the main causes of death. The flavonoid with a hydroxyl group (OH) on its C-3 position is called flavonol. 3-hydroxyflavone is the structural skeleton for flavonol whichhas been reported not only to inhibit cell proliferation in breast cancer, prostate cancer and colon cancer, but also have the effect on the inhibition for human osteosarcoma cells. The study has been designed to investigate 3-hydroxyflavone if it may inhibit the invasion and metastasis of 143B human osteosarcoma cells. First, after injection of cancer cells into immunodeficient mice (BALB/cAnN.CgFoxnlnu/ CrlNarl), the change of body weight in mice, the tumor growth of body surface, and tumor size were observed. Second, for comparing the experimental group fed with 3-hydroxyflavone (tube feeding) to the unfed control group, in vivo tumor growth and cancer metastasis were observed further by in vivo image analysis system (IVIS).These results have showed that 3-hydroxyflavone could inhibit the tumor size of osteosarcoma, However, no metastasis occurred during the observation of metastasis for cancer cells. Further using 143B cells to explore if 3-hydroxyflavone may inhibit cell invasion and metastasis by MTT assays, modified Boyden chamber assay, zymography assay and cell wound healing assay, which have found that 3-hydroxyflavone is no cytotoxicity, is able to reduce the expression level of MMP-2 in bone cancer cells, and has the effect of inhibition on the abilities of cell mobility and invasion. Based on these experimental results above, which have suggested that 3-hydroxyflavone may have the inhibitory effecton the invasion bility of human osteosarcoma cells, however, further long-term follow-up in animal studies and protein analysis have to be performed in order to obtain more favorable evidences for application of clinical therapy.

並列關鍵字

Osteosarcoma ； flavonoid ； flavonol

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