Different Mechanisms of Berberine-induced Anti-proliferation Effects in Triple Negative Breast Cancer MDA-MB-231 and MDA-MB-468 Cell Lines

• Main Authors: Yi-Hsiu Tsai, 蔡易修
• Other Authors: Jen-Hwey Chiu
• Format: Others
• Language: en_US
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/74561788716606686662

碩士 === 國立陽明大學 === 傳統醫藥研究所 === 105 === Breast cancer is the leading cause of cancer death among females worldwide and ranks 4th in Taiwan. Among the different subtypes, triple negative breast cancer (TNBC), lack of estrogen-receptor (ER), progesterone-receptor (PR) and human epidermal growth factor receptor 2 (HER2) expressions, is characterized by its higher aggressiveness, higher recurrence rate, and poorer prognosis. Although there are emerging targeted therapies developed, the treatment of TNBC remains difficult clinically. Berberine, one of the main active components of traditional Chinese medicine (TCM), Huanglian (Coptidis rhizome), it is a kind of isoquinoline alkaloids. Berberine was shown to be effective in inhibiting cell proliferation and promoting apoptosis in various cancer cells. However, the effects of berberine on TNBC cell lines remain unclear. The aim of this study was to investigate the cell inhibition effects of berberine in triple negative breast cancer (TNBC) cells. By using human TNBC cell lines MDA-MB-231 and MDA-MB-468 as in vitro model, anti-proliferative effects of berberine on TNBC cells were elucidated by MTT assay, trypan blue exclusion assay, and clonogenic assay. Cell necrosis, apoptosis, and autophagy were analyzed by cell morphology, flow cytometry, immunofluorescence staining and Western blot for LC3Ⅰ/Ⅱ, respectively. Cell growth-related signaling pathway AKT/ERK/p38 and cell cycle kinase complexes proteins expression were analyzed by Western blot. In vivo MDA-MB-231 bearing xenografted nude mice was used to study the combination effects of berberine and doxorubicin (DOX), compared to DOX alone group. Based on MTT assay and clonogenic assay, berberine concentration-dependently suppressed cell proliferation in MDA-MB-468 (0, 3, 6, 12 μM) and MDA-MB-231(0, 6.25, 12.5, 25 μM), but this inhibitory effect was not through cell necrosis, apoptosis, and autophagy. Cell cycle analysis showed that an increased S/G2/M fraction and a decreased G0G1 phase in berberine-treated MDA-MB-231 cells, while a decreased S/G2/M phase fraction was noticed in berberine-treated MDA-MB-468 line. By Western blot, berberine not only decreased the activation of AKT/ERK but also decreased the expression of Cyclin A and CDK1 in MDA-MB-231, while berberine increased the activation of p38 and decreased the expression of Cyclin D and CDK4 in MDA-MB-468. Moreover, a combination of berberine + DOX had a more tumor-suppressive effect than DOX alone group. Our results demonstrate that the anti-proliferation effects of berberine were through the different mechanism in TNBC MDA-MB-231 and MDA-MB-468 cell lines, which provides the potential of berberine in personalized TNBC therapy.