Department of Biochemistry and Molecular Biology, Medical University of Lublin, 20-093 Lublin, Poland.
Department of Pathophysiology, Medical University of Lublin, 20-090 Lublin, Poland.
Int J Mol Sci. 2022 Jun 9;23(12):6458. doi: 10.3390/ijms23126458.
Breast cancer (BC) is a heterogeneous disease with different intrinsic subtypes. The most aggressive subtype of BC-triple-negative breast cancer (TNBC) is characterized by high heterogeneity and metastasis rate, poor prognosis and lack of therapeutic targets due to the absence of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2. Targeted therapies have been approved for many other cancers and even other subtypes of BC, but treatment options for TNBC are still mainly limited to chemotherapy. Therefore, new, more effective treatment regimens are needed. Combined chemotherapy with two or more active agents is considered a promising anti-neoplasm tool in order to achieve better therapeutic response and reduce therapy-related adverse effects. The study demonstrated an antagonistic effect commonly used in TNBC therapy cytostatic drug-paclitaxel (PAX) and sirtuin inhibitor: cambinol (CAM) in BT-549, MDA-MB-468 and HCC1937 TNBC cell lines. The type of pharmacological interaction was determined by a precise and rigorous pharmacodynamic method-isobolographic analysis. The cytotoxic and anti-proliferative effects of CAM used alone or combined with PAX were determined utilizing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-bromo-2'-deoxyuridine (BrdU) assays, respectively. Induction of apoptosis in TNBC cell lines after PAX and CAM treatment applied individually or in combination was determined by flow cytometry (FACS) as a number of cells with active caspase-3. It has been observed that both agents used separately inhibit cell proliferation and induce apoptosis; however, applying them in combination ameliorated antiproliferative and pro-apoptotic effects in all analyzed TNBC cell lines. Our results demonstrate that CAM and PAX used in combination act antagonistically, limiting anti-cancer efficacy and showing the importance of preclinical testing.
乳腺癌（BC）是一种具有不同内在亚型的异质性疾病。最具侵袭性的 BC 亚型——三阴性乳腺癌（TNBC）由于缺乏雌激素受体、孕激素受体和人表皮生长因子受体 2，具有高度异质性和转移率、预后不良以及缺乏治疗靶点等特点。许多其他癌症甚至其他 BC 亚型都已经批准了靶向治疗，但 TNBC 的治疗选择仍然主要限于化疗。因此，需要新的、更有效的治疗方案。联合两种或更多种活性药物的化疗被认为是一种有前途的抗肿瘤工具，以实现更好的治疗反应并减少与治疗相关的不良反应。该研究在 BT-549、MDA-MB-468 和 HCC1937 TNBC 细胞系中证明了常用的 TNBC 治疗细胞抑制剂紫杉醇（PAX）和组蛋白去乙酰化酶抑制剂 cambinol（CAM）之间存在拮抗作用。通过精确和严格的药效学方法——等效应线分析来确定药物相互作用的类型。单独使用或联合使用 CAM 的细胞毒性和抗增殖作用分别通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）和 5-溴-2'-脱氧尿苷（BrdU）测定来确定。用流式细胞术（FACS）检测 PAX 和 CAM 单独或联合处理后 TNBC 细胞系中凋亡的诱导，作为具有活性 caspase-3 的细胞数量。已经观察到两种药物单独使用时均能抑制细胞增殖并诱导凋亡；然而，联合使用时在所有分析的 TNBC 细胞系中改善了抗增殖和促凋亡作用。我们的结果表明，CAM 和 PAX 联合使用时表现出拮抗作用，限制了抗癌效果，并表明临床前测试的重要性。
