- 學位論文
查爾酮衍生物抑制肺癌機制之探討
Study of Anti-Lung Cancer Activity by Chalcone Derivatives
摘要
近年來多種天然草藥陸續被證實具有抗癌能力，查爾酮 (chalcone) 是植物合成黃酮類的主要前趨複合物，研究指出它具有抗發炎、抗菌、抗瘧以及抗癌等功能，但chalcone對於肺癌之相關研究不多。本研究主要在於探討查爾酮衍生物對於肺癌的抑制作用。MTS assay分析發現查爾酮衍生物2-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone對於癌細胞較具毒殺效果，研究發現組織中端粒酶活性喪失則其預後較佳，因此抑制端粒酶活性已成為最新的抗癌策略之一。本研究以TRAP assay分析發現2’-Hydroxy-2, 3, 4’, 6’, tetramethoxychalcone可以降低肺癌A549細胞端粒酶活性，以RT-PCR和Western blot analysis分析hTERT mRNA和蛋白表現量，也有減少的趨勢，以Luciferase Reporter Assay分析發現，2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone具有抑制h.TERT啟動子活性的作用。另外，以細胞內鈣離子螯合劑BAPTA-AM和TRAP assay分析發現，鈣離子可以恢復被2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone抑制之端粒酶活性，以鈣離子染劑Fluo-3AM和共軛焦顯微鏡觀察發現2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone會使細胞內鈣離子濃度上升，結果顯示鈣離子可能參與2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone抑制端粒酶活性之作用。而以Boyden chamber、gelatin zymography和wound healing assay分析發現，查爾酮衍生物對於抑制肺癌細胞的轉移，並無明顯的作用。最後以soft agar和動物腫瘤實驗分析，發現2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone可以減緩腫瘤初期的生長。綜合以上實驗結果，查爾酮衍生物2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone可以抑制肺癌細胞之端粒酶活性和生長。將來可發展為抑制端粒酶活性和腫瘤生長之臨床用藥。
關鍵字
查爾酮並列摘要
Many clinically successful anticancer drugs are themselves either natural products or have been developed from Chinese herbs. Chalcones are the intermediate precursors for all flavonoids. Chalcones and their derivatives have been shown to possess a wide range of biological functions including anti-inflammatory, anti-bacterial, anti-malarial, anti-cancer. We found 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone is preferentially toxic to cancer cell lines in comparison with normal lung cell lines. Previous studies have suggested that the loss of telomerase activity is a favotable prognostic marker. Therefore, telomerase has become an attractive target for anti-cancer therapeutic interventions. In this study, we demonstrated that 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone suppressed telomerase activity expression in A549 cells. 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone also repressed hTERT promoter activity, resulted in decreased hTERT mRNA expression. Treatment of intracellular calcium chelator BAPTA-AM neutralized the inhibitory effect of 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone on telomerase activity. Using Fluo-3AM for Ca2+ detection, we proved that 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone induced increased intensity of intracellular calcium signals. These data indicate that calcium might participate in the suppression of telomerase activity. In addition, we also evaluated the effect of chalcone, flavonoid and isoflavonoid derivatives on A549 cell migration by Boyden chamber, gelatin zymography and wound healing, however, no inhibitory effect on cell migration was observed. Finally, the anti-tumor growth effect of 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone was evident in its inhibitory growth of lung tumor A549 cells in nude mice model and in soft agar, chalcone derivative 2’-Hydroxy-2, 3, 4’, 6’-tetramethoxychalcone inhibits telomerase activity in lung cancer cells and tumor growth, provides a rationale for the further development of this class of compounds as novel cancer chemotherapeutic agents.
並列關鍵字
Chalcones參考文獻
延伸閱讀
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