茶黃素-3,3\'-雙沒食子酸酯抑制人類骨肉瘤細胞侵入與移動之研究

骨肉瘤 (Osteosarcoma，OS)是一種主要發生於兒童、青少年及年輕成人 (10-30歲)之罕見癌症，因初期症狀不明顯難被診斷出來及預後不佳復發等原因而致使許多患者死於癌肺部轉移，因此，抑制骨肉瘤癌細胞轉移為治療的一大突破的重點，在臨床和實驗證據顯示，尿激酶纖溶酶原啟動劑 (urokinase-type plasminogen activators，u-PA) 和基質金屬蛋白酶 (matrix metalloproteinases， MMPs) 表現升高與癌症進展、轉移和患者生存期縮短有關，而在骨肉瘤中MMP-2及u-PA與其癌症進展、轉移和預後不佳皆起著至關重要的作用，因此，可將抑制MMP-2及u-PA作為治療目標進行研究，而茶黃素-3,3'-雙沒食子酸酯 (theaflavin-3,3'-O-digallate，TF3) 其有抑制癌細胞增殖、誘導癌細胞凋亡、抑制血管新生、合併抗癌藥物增強其抑制癌細胞效果等多種功能，然而，它對骨肉瘤癌症轉移的影響仍然有許多未知。本實驗針對TF3在骨肉瘤細胞中的抗轉移效果進行研究，結果顯示，以MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay分析TF3處理143B和U2OS細胞之細胞存活率，數據顯示TF3 60 μM處理24小時對於143B和U2OS具有細胞毒性，再以Wound healing assay、Boyden chamber進行細胞的遷移、移動和侵襲等能力分析，結果顯示TF3可抑制細胞的遷移、移動和侵襲等能力的效果有隨著其濃度上升而增加，且以Zymography 分析MMP-2 和u-PA的分泌量的結果中也觀察到同樣有抑制效果，而在細胞群落分析中以TF3 10 μM處理的143B和U2OS也有很好的抑制效果。結合上述結果確認TF3可有效抑制骨肉瘤的轉移。

Osteosarcoma is a non-common cancer which mostly occur in children, teenagers, and young adults between the ages of 10 to 30. Due to its unobvious primary symptoms, low suspicion by physicians, poor prognosis, and relapsing of cancer which might present with remitting symptoms; the increased mortality rate caused by metastasis to the lungs is common. Thus, suppression of cancer cells metastasis is a major-to-breakthrough in osteosarcoma treatment. There’s clinical and research evidence indicates that elevation of urokinase-type plasminogen activator (u-PA) and matrix metalloproteinases (MMPs) levels are associated in cancer progression, metastasis and curtailed survival of patients; whilst in osteosarcoma, matrix metalloproteinase-2 (MMP-2) and u-PA are thence a key roles correlated in its tumour cell progression, metastasis and prognosis. Therefore, research in inhibition of MMP-2 and u-PA could be set as a treatment goal. Theaflavin-3,3'-O-digallate (TF3) reportedly have the ability to directly suppress cancer cells growth, induce cancer cells apoptosis, inhibit angiogenesis, combined with chemotherapy drugs providing synergistic effect in enhancing repression of cancer cells, etc. However, the function of TF3 towards osteosarcoma is still remain unknown. Experiments are conducted to study the anti-metastasis effect of TF3 in osteosarcoma cells. After treating 143B cells and U2OS with TF3, an MTT assay is conducted to analysis the cell viability. The results demonstrated that TF3 treatment (60 μM) for 24hr exhibited cytotoxicity on the 143B and U2OS cell lines. Wound healing assay and Boyden chamber is later conducted to analysis the cell motility, cell migration, and cell invasion. Results showed that TF3 could suppress cell motility, cell migration and cell invasion in a dose-dependent manner. Same suppression potential is observed in MMP-2 and u-PA activity using Zymography analysis. TF3 treatment (10 μM) on the 143B and U2OS cell colony also present a great suppression results. Taken together, these data showed reveal that TF3 have a great potential in suppressing osteosarcoma metastasis.