Laboratório de Investigação Molecular do Câncer (LIMC), Faculdade de Medicina de São José do Rio Preto - FAMERP, Av. Brigadeiro Faria Lima, 5416, 15090-000 São José do Rio Preto, SP, Brazil; Postgraduate Program in Health Sciences, Faculdade de Medicina de São José do Rio Preto - FAMERP, Av. Brigadeiro Faria Lima, 5416, 15090-000 São José do Rio Preto, SP, Brazil.
Postgraduate Program in Health Sciences, Faculdade de Medicina de São José do Rio Preto - FAMERP, Av. Brigadeiro Faria Lima, 5416, 15090-000 São José do Rio Preto, SP, Brazil; Universidade Paulista - UNIP, São José do Rio Preto, SP, Brazil.
Life Sci. 2023 Jul 1;324:121708. doi: 10.1016/j.lfs.2023.121708. Epub 2023 Apr 20.
Breast cancer (BC) presents high mortality rate and about 25-46 % have mutation in the PIK3CA gene. Alpelisib is a PI3K inhibitor that acts on p110α, which is a subunit of the PI3K protein. The melatonin shown important anti-neoplastic effects and may increase the effectiveness of chemotherapy. This study evaluated the synergistic action of Alpelisib and Melatonin in BC lines carrying the H1047R mutation in PIK3CA, relative to the cellular dynamics and the PI3K/AKT/mTOR pathway.
MDA-MB-468 (triple-ernegative), MDA-MB-453 (H1047R PIK3CA, HER2+) and T-47D cells (H1047R PIK3CA, ER+/PR+) were divided into four treatment groups: control; Melatonin (1 mM); Alpelisib (1 μM); and Alpelisib (1 μM) + Melatonin (1 mM). Cell viability and migration were investigated using the MTT assay and Transwell assay, respectively. Protein expression of PI3K, p-AKT, mTOR, HIF-1α, and caspase-3, was verified using immunocytochemistry.
MTT assay revealed that MDA-MB-453 and T-47D showed reduction in cell viability in all groups, especially in the MDA-MB-453 treated with Melatonin + Alpelisib. MDA-MB-468 presents reduction in cell migration only with Melatonin, while in the lines with mutation, the treatment of Melatonin + Alpelisib caused inhibition of cell migration. PI3K, p-AKT, mTOR and HIF-1α were inhibited after treatment with Melatonin + Alpelisib in MDA-MB-453 and T-47D lines. The expression of caspase-3 increased in all groups in MDA-MB-453 and T-47D cells, being the increase more pronounced in the Melatonin + Alpelisib group.
These results indicate that the combined use of Melatonin and Alpelisib may be more effective in inhibiting BC in women carrying the PIK3CA gene mutation than either treatment alone.
乳腺癌（BC）死亡率高，约 25-46%存在 PIK3CA 基因突变。阿培利司是一种针对 p110α 的 PI3K 抑制剂，p110α 是 PI3K 蛋白的一个亚单位。褪黑素具有重要的抗肿瘤作用，并可能提高化疗的效果。本研究评估了阿培利司和褪黑素在携带有 PIK3CA H1047R 突变的 BC 细胞系中的协同作用，相对于细胞动力学和 PI3K/AKT/mTOR 通路。
MDA-MB-468（三阴性）、MDA-MB-453（PIK3CA H1047R，HER2+）和 T-47D 细胞（PI3CA H1047R，ER+/PR+）分为四组：对照组；褪黑素（1mM）；阿培利司（1μM）；阿培利司（1μM）+褪黑素（1mM）。用 MTT 法检测细胞活力，用 Transwell 法检测细胞迁移。用免疫细胞化学法检测 PI3K、p-AKT、mTOR、HIF-1α 和 caspase-3 的蛋白表达。
MTT 法显示 MDA-MB-453 和 T-47D 组在所有组中均表现出细胞活力降低，尤其是 MDA-MB-453 组中褪黑素+阿培利司组降低更为明显。MDA-MB-468 组仅用褪黑素表现出细胞迁移减少，而在突变株中，褪黑素+阿培利司治疗导致细胞迁移抑制。在 MDA-MB-453 和 T-47D 细胞中，褪黑素+阿培利司处理后 PI3K、p-AKT、mTOR 和 HIF-1α 被抑制。MDA-MB-453 和 T-47D 细胞中各组 caspase-3 的表达均增加，褪黑素+阿培利司组增加更为明显。
这些结果表明，与单独使用阿培利司或褪黑素相比，联合使用褪黑素和阿培利司可能更有效地抑制携带 PIK3CA 基因突变的女性乳腺癌。
