先前實驗室發現強力黴素(Doxycycline, DOX)和強力黴素衍生物(TMC-1)可以抑制SCC15細胞之基質金屬蛋白酶MMP-2及MMP-9的表現。TGF-β1是一種多功能的免疫調節劑，癌症發生前，TGF-β1抑制腫瘤發展和生長以及促進細胞凋亡的方式來減緩癌細胞的增生，然而在腫瘤細胞中，TGF-β1會促進癌細胞生長及轉移，尤其較為後期的腫瘤組織中，往往有更大量的TGF-β1表現。本篇研究探討SCC25細胞受TGF-β1誘導的時間長短對MMPs的影響。由明膠蛋白酵素電泳的結果顯示，TGF-β1誘導5天時，MMP-2的表現量會是誘導1天時的3倍，而誘導1天至3天時，MMP-9的表現量相當少，誘導4天及5天時出現了明顯的表現。我們亦探討DOX、DOX衍生物(TMC-1)和茶籽萃取物(NCF)在SCC25受TGF-β1誘導1天及5天時抑制MMP-2及MMP-9的能力。經TGF-β1誘導1天時，DOX和TMC-1抑制MMP-2的IC50分別為6.879和2.146 μg/mL。誘導5天時，抑制MMP-2及MMP-9的IC50分別為4.623, 3.053 μg/mL及6.431, 3.585 μg/mL。此外，NCF經誘導1天及5天時，IC50為2868及251 μg/mL。TGF-β1誘導時間長時，NCF抑制MMP-2及MMP-9的效果更加顯著。總而言之，DOX、TMC-1和NCF皆能夠抑制SCC25細胞的MMP-2及MMP-9表現。

Our previous studies have shown that Doxycycline (DOX) can inhibit the expression level of MMP-2 and MMP-9 in squamous-cell carcinoma cell line (SCC15) and the inhibition is possibly through TGF-β1/Smad signaling pathway. In this report, we used TGF-β1 to induce the expression of MMPs in SCC25 cells from 1 to 5 days and the expression level of MMPs in the culture medium were analyzed by gelatin zymography. Our results showed that MMP-2 was detected in Day 1 and it was gradually induced up to three fold in Day 5. On the other hand, MMP-9 was not detected before Day 3 and it was detected in Day 4 and 5. This result indicates that MMP-2 is the major MMPs in SCC-25 cell line and its expression can be induced by TGF-β1. In addition, MMP-9 is not detectable in SCC25 cell line but it was detectable after 4 days of TGF-β1 induction. We then tested if the expression level of MMP-2 and MMP-9 after 1 day and 5 days induction by TGF-β1 can be inhibited by DOX, DOX analogs (TMC-1), and non-catechin flavonoids (NCF) from Camellia sinensis (L.) O. Kuntze seed. Our results showed that both DOX and TMC-1 can inhibit the expression of MMP-2 in Day 1 with IC50 of 6.879 and 2.146 μg/mL and Day 5 with IC50 of 4.623 and 3.053 μg/mL. They can also inhibit the expression of MMP-9 in Day 5 with IC50 of 6.461 and 3.585 μg/mL. In addition, NCF was able to inhibit the expression of MMP-2 in Day 1 and Day 5 with IC50 of 2868 and 251 μg/mL. It is interesting to see the difference of IC50 in Day 1 and Day 5. In conclusion, MMP-2 is the major MMPs in SCC25, its expression level can be induced by TGF-β1 and inhibited by DOX, TMC-1 and NCF.

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