野生苦瓜(Momordica Charantia Linn. var. abbreviata Seinge)是亞洲常見的蔬菜和傳統藥物。在最近的研究中，野生苦瓜中的三萜類化合物已在多種癌細胞中顯示出抗腫瘤活性。 3β，7β，25-trihydroxycucurbita-5，23(E)-dien-19-al (TCD) ，一種從野生苦瓜中分離出來的三萜類化合物在乳腺癌細胞研究中TCD會誘導其細胞凋亡。因此我們進一步想了解TCD 對各種癌細胞株的生長抑制情形。
首先我們利用MTT法來檢測不同濃度TCD處理後的細胞生長活性，結果顯示TCD可以抑制多種癌細胞株的生長，但不會影響肺上皮細胞(MCR5)。另外我們亦觀察到TCD抑制子宮頸癌細胞株(HeLa)的生長最為顯著，所以我們進一步則想探討TCD誘導子宮頸癌的死亡的分子機制。我們利用傷口癒合試驗來觀察癌細胞的遷移情形，結果發現經10μg/ml TCD處理後，癌細胞的遷移能力明顯地受到抑制。接著利用流式細胞儀觀察TCD處理後細胞週期的變化狀況，我們亦發現TCD可誘導癌症細胞停留在G1期。進一步釐清TCD誘導細胞死亡是否經由細胞凋亡(apoptosis)，我們進行Annexin V 染色，結果顯示細胞凋亡的數目會隨著TCD的濃度而增加，同時我們亦會分析調控細胞凋亡的蛋白分子表現量。
整體來說，TCD可抑制多種癌細胞株的生長，而進一步在子宮頸癌的研究，發現TCD會誘導細胞停留在G1及細胞凋亡。這些結果顯示TCD或許可作為未來抗癌藥物之一。
Wild bitter melon (Momordica charantia Linn. var. abbreviata Seinge) is a common vegetable and traditional medicine in Asia. In recent research, 3β,7β,25-trihydroxycucurbita-5,23(E)-dien-19-al (TCD), a triterpenoid was isolated from wild bitter melon has showed anti-tumor activity in various cancer cells. Recently, TCD can induce breast cancer cells apoptosis. Therefore, we wanted to understand the growth inhibition status of TCD on various cancer cell lines.
First, we used the MTT method to detect the cell growth activity after treatment with different concentrations of TCD. The results show that TCD can inhibit the growth of various cancer cell lines, but does not affect lung epithelial cells (MCR5). In addition, we observed that TCD inhibited the growth of cervical cancer cell line (HeLa) most significantly, so we further wanted to explore the molecular mechanism of TCD-induced cervical cancer death. We used wound healing assay to observe the migration of cancer cells, and found that the migration ability of cancer cells was significantly inhibited after 10μg/ml TCD treatment. Then use flow cytometry to observe the changes of cell cycle after TCD treatment, we found that TCD can induce cells to stay in G1 phase. To further clarify whether TCD-induced cell death is via apoptosis, we performed Annexin V staining. The results showed that apoptotic cells increased with the concentration of TCD, and accompanied the activation of apoptosis-related protein molecules.
Overall, TCD inhibited the growth of a variety of cancer cell lines. Further research on cervical cancer found that TCD could induce cells to stay in G1 and apoptosis. These results suggest that TCD may be one candidate of the future anticancer drugs.
