| 摘要: | 膀胱癌是泌尿道最常見的癌症之一，男性罹患癌症機率比女性多三倍。每年死於膀胱的患者數量都在增加。由於癌症具有轉移的能力，因此治療癌症並不容易。儘管手術技術的進步和全身化療的改善，膀胱癌的5年存活率沒有改變。為了要克服上述的所有困難，所以尋找抗癌和抗轉移的新藥是迫切需要的。在這個研究中，槲皮素˗鋅復合 (quercetin-Zn) 被發現，它不僅具有濳在的抗癌特性，而且能夠有效抑制膀胱癌細胞的遷移、侵襲和轉移。槲皮素(quercetin) 是最具生物活性的黃酮類合物之一，有在許多水果，蔬菜和飲料。由於其抗癌，抗氧化和抗發炎效果，因此槲皮素被作為化學藥物研究應用在各種癌症。然而，槲皮素很難被吸收到生物體內，因此導致生物利用度差。有趣的是，槲皮素能與金屬配位以形成複合物並增強其生物利用度，並改變體內傳遞槲皮素的方式。槲皮素˗鋅復合的抗癌活性已被證明，因此，槲皮素˗鋅複合物是癌症治療藥物中具有前景的藥物。然而，槲皮素˗鋅對腫瘤細胞的抑制作用並沒有完全瞭解。槲皮素屬於廣泛類型的酚類化合物，酚類化合物已被報導作為對癌症侵襲和轉移的潛在抑制作用，因此認為槲皮素˗鋅複合物也具有這種性質。在這項研究中，合成了槲皮素˗鋅，其化學性質顯示在UV-VIS，FT-IR，1H-NMR，表明通過3-OH 和C4=0 基團的結合位點。比較了槲皮素˗鋅，槲皮素和鋅離子對BFTC-905和NIH-3T3 細胞的抗癌作用。對BFTC-905細胞的細胞毒性測試結果表明，槲皮素˗鋅殺傷作用比單獨槲皮素高2.8倍。且在NIH-3T3細胞上也發現有相同作用，槲皮素˗鋅對NIH-3T3細胞的細胞毒性比單獨使用槲皮素較高。此外鋅離子對BFTC-905和NIH-3T3細胞都是無毒的。進一步的研究發現了槲皮素˗鋅抗轉移作用在BFTC-905細胞在非細胞毒性濃度（0-50μM）下。在此範圍中，用槲皮素-鋅處理的BFTC-905細胞通過傷口癒合測試明顯地顯示濃度上升時細胞遷移能力降低。基於侵犯測試結果經槲皮素-鋅複合物處理24 h後，顯著侵入膜下側的BFTC-905細胞與對照組相比降低（p值<0.05），提示BFTC-905細胞用槲皮素-鋅複合物處理顯示顯著減少侵犯能力。此外為了研究槲皮素-鋅複合物的抗轉移機制，檢測了AKT和MT1-MMP的表達。 我們發現，p-AKT和MT1-MMP水平的表達下調槲皮素鋅處理的BFTC-905細胞。 最重要的，這些研究提供更好的方式去了解槲皮素-鋅複合物的分子途徑抑制轉移和侵犯可能是治療膀胱癌有用的方式。;Bladder cancer is one of the most frequent cancer of urinary tract and it’s nearly three time more common in men than women. The patients died of bladder cancer increase every year. It could be the effective treatments for tumors are not easy to achieve due to existence of metastases, tumor metastases mainly leads to high mortality rate of bladder cancer patients. In spite of advances in surgical techniques and improvements of systemic chemotherapies, there has been no change in the 5-year survival rate of bladder cancer. To overcome all these difficulties above, looking for a new drug is pressing need, which should be antitumor and anti-metastasis activities. In this research, quercetin-zinc (quercetin-Zn) was found that, it possessed not only potent anticancer activity, but was also able to effectively inhibit migration, invasion and metastasis of human bladder cancer cells (BFTC-905). Quercetin is one of the most bioactive flavonoids compound and present in many fruits, vegetables, beverages. Due to its anticancer, anti-oxidant and anti-inflammation activity, quercetin has been studied extensively as a chemoprevention agent in variety cancer models. However, quercetin was found to be difficult absorbed into the body, thus resulted in poor bioavailability. Interestingly, quercetin is the ideally compound for coordinate with metal to form a complex compound and enhance its biological activity with increasing bioavailability and also change the way deliver quercetin in vivo. The anticancer activity of quercetin-Zn complex has been demonstrated, therefore, quercetin-Zn is a promising complex in developing new medicine for cancer treatment. However, many aspects of the inhibitory effects of quercetin-Zn to tumor cells are not still completely understood. Quercetin is belong to extensive class of phenolic compounds, phenolic compounds have been report as a potential inhibitory effect on cancer invasion and metastasis, thus, it is believed that quercetin-Zn complex also own this property. In this work, quercetin-Zn was synthesized and its chemical properties were characterized by UV-VIS, FT-IR, 1H NMR, which showed that binding site via 3-OH and C4=O group. The anticancer effects of quercetin-Zn, quercetin and zinc ion on BFTC-905 and NHI-3T3 cells were compared. The result of cell viability on BFTC-905 cells demonstrated that killing effect was 2.8 time higher than quercetin alone. However, this effect was also found on NIH-3T3 cells, cytotoxic effect of quercetin-Zn on NIH-3T3 cells was shown to be higher than quercetin alone. In contrast, zinc ion was found to be non-toxic to both BFTC-905 and NIH-3T3 cells. Further investigation revealed the anti-metastasis effect of quercetin-Zn on BFTC-905 cells under non-cytotoxic concentration. In non-cytotoxic concentration range (0-50µM), BFTC-905 cells that were treated with quercetin-Zn significantly exhibited a concentration-dependent reduction in cell migration by wound healing assay. Based on the invasion assay result, after 24 h treatment with quercetin-Zn complex, the amount of BFTC-905 cells that invaded to the lower side of the membrane markedly reduced compare with control (p-value<0.05), suggested that BFTC-905 cells were treated with quercetin-Zn complex display significant reduction in invasive capacity. Furthermore, in order to investigate the anti-metastasis mechanism of quercetin-Zn complex, the expression of AKT and MT1-MMP were examined. We found that, the expression of p-AKT and MT1-MMP level were down-regulated in quercetin-Zn-treated BFTC-905 cells. Importantly, these findings provide better understanding for the molecular pathway of quercetin-Zn complex on suppression of migration and invasion which might be useful as a therapeutic strategy to treat human bladder cancer. |
