探討洛神花青素抗乳癌細胞之作用及其分子機制

Studies of the anti-cancer effect and mechanisms of Hibiscus anthocyanins in human breast cancer cells

姚芳儀(Fang-Yi Yao)

指導教授 ： 王朝鐘 ； 張雲菁

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2011年)

摘要

在全球，女性乳癌的好發率高，大多數乳癌患者常因復發或轉移而死亡，且先前我們的研究已經證實洛神花青素(Hibscus anthocyanins ,HAs)具有抗氧化、抗發炎、預防心血管病變及抗血癌、大腸癌等功效，但在乳癌上是否具有抑制作用尚未清楚。因此，本研究擬探討洛神花青素抗乳癌作用及其分子機轉。首先，本研究選用人類乳癌細胞株(MCF-7及MDA-MB-231細胞)進行實驗，利用MTT、流式細胞儀、JC-1染色、H2DCF-DA染色、西方點墨法等方法分析，結果發現經洛神花青素處理後，MCF-7細胞的IC50約為2.3 mg/ml，MDA-MB-231細胞則約為3.4 mg/ml，因MCF-7細胞對洛神花青素敏感度較高，故後續以MCF-7細胞為研究材料。進一步觀察處理洛神花青素後之細胞形態，細胞呈現皺縮、有空泡產生及細胞死亡等現象。另外，將MCF-7細胞處理低劑量的洛神花青素(0, 0.25, 0.5, 0.75 mg/ml)後，促進細胞週期停滯在G0/G1期，使細胞增生受到抑制；而當細胞處理高劑量的洛神花青素後(0, 1, 2, 3 mg/ml)，則會促進MCF-7細胞死亡，死亡方式並非進行凋亡(apoptosis)或自噬(autophagy)，而是透過增加細胞內ROS表現量，刺激粒線體通透性轉換增加，使粒線體內膜上的 permeability transition pore持續打開，造成細胞壞死(necrosis)。綜合以上結果，洛神花青素有效地使乳癌細胞死亡，期望未來在乳癌的治療上能有極佳的輔助效果。

關鍵字

洛神花青素 ； 乳癌 ； 細胞週期停滯 ； 細胞壞死

並列摘要

Breast cancer is a high incidence cancer in woman worldwide, and the causes of death are cancer relapse or metastasis in most of patients. Our previous reports have demonstrated that the Hibscus anthocyanins (HAs) is potent provided with anti-oxidation, anti-inflammation, preventing cardiovascular disease, and anti-cancer including leukemia and colon cancer. However, the anti-breast cancer effect of the HAs is not identifying. Thus, we investigated whether HAs is ability to anti-breast cancer and the mechanisms. First, our experiments chose two human breast cancer cells (MCF-7 and MDA-MB-231), and we assayed using MTT assay, flow cytometery, JC-1 staining, H2DCF-DA staining, western blotting assay, and so forth. Our results showed that the IC50 is about 2.3 mg/ml in MCF-7 cells and the IC50 is about 3.4 mg/ml in MDA-MB-231 cells. The MCF-7 cells are more sensitive than MDA-MB-231when cells are treated with HAs. Therefore, we utilized the MCF-7 cells to future study at our subsequently experiments. In addition to we observed the cell morphology, and found that the HAs induced MCF-7 morphologic shrink, forming vesicle, and death. We also discovered that the low doses HAs (0, 0.25, 0.5, 0.75 mg/ml) promoted MCF-7 cell cycle arrest at G0/G1 phase and significantly decreased cell proliferation. Furthermore, the high doses HAs (0, 1, 2, 3 mg/ml) increased intracellular ROS levels, augmented mitochondrial permeability transition, and the permeability transition pore in mitochondrial inner membranes are constantly opening. Hence, the high doses HAs induced MCF-7 cells death by necrosis but not apoptosis or autophagy. In brief, the HAs are extremely potential for inducing breast cancer cells death. We have good expectations of the HAs greatly having adjuvant therapy for breast cancer in future.

並列關鍵字

HAs ； human breast cancer cells ； cell cycle arrest ； necrosis

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