Department of Thyroid and Breast Diseases, Jincheng People's Hospital, Jincheng, Shanxi, China.
Bioengineered. 2022 Apr;13(4):10944-10955. doi: 10.1080/21655979.2022.2066758.
Breast cancer is characterized by high morbidity and mortality. GDC-0941 is a PI3K inhibitor with oncogenic effects in breast cancer. However, certain breast cancer cells are insensitive to GDC-0941. Hence, the mechanism of GDC-0941 in breast cancer resistance was investigated in this study. Breast cancer cell lines BT-474, MCF7, Hs-578-T, MDA-MB-231, MDA-MB-453, and MDA-MB-468 were cultured in different medium and then treated with 100 or 500 nM GDC-0941, 100 nM OSU-T315, or TNF-α antibody. Moreover, ILK and shILK were transfected into cells. The half maximal inhibitory concentrations (IC50) for GDC-0941 were detected using CCK-8 assay. The levels of ILK, AKT, PDK1, S6, and p70S6K expression were detected using western blotting and qPCR. In addition, the mouse model of breast cancer was constructed to measure the tumor size, volume, and weight. The results showed that GDC-0941 decreased cell survival rate, downregulated the phosphorylation of AKT, S6, and p70S6K, and promoted the expression of ILK, while it had little effect on PDK1 expression. GDC-0941 inhibited the increases in p-AKT, p-S6, and p-p70S6K caused by ILK overexpression and promoted ILK knockdown-induced reduction of p-AKT, p-S6, and p-p70S6K. In addition, the combination of OSU-T315 and GDC-0941 decreased p-AKT, p-S6, and p-p70S6K level, tumor volume, and tumor weight. GDC-0941 promoted ILK expression by upregulating TNF-α level. Taken together, GDC-0941 increased ILK level by upregulating TNF-α, thus affecting AKT expression and the sensitivity of breast cancer cells to GDC-0941.
乳腺癌具有高发病率和高死亡率的特点。GDC-0941 是一种具有致癌作用的 PI3K 抑制剂，可用于乳腺癌的治疗。然而，某些乳腺癌细胞对 GDC-0941 不敏感。因此，本研究旨在探讨 GDC-0941 在乳腺癌耐药中的作用机制。将乳腺癌细胞系 BT-474、MCF7、Hs-578-T、MDA-MB-231、MDA-MB-453 和 MDA-MB-468 分别在不同的培养基中培养，然后用 100 或 500nM GDC-0941、100nM OSU-T315 或 TNF-α 抗体处理，同时转染 ILK 和 shILK。使用 CCK-8 检测 GDC-0941 的半抑制浓度（IC50）。使用 Western blot 和 qPCR 检测 ILK、AKT、PDK1、S6 和 p70S6K 的表达水平。此外，构建乳腺癌小鼠模型以测量肿瘤的大小、体积和重量。结果表明，GDC-0941 降低了细胞存活率，下调了 AKT、S6 和 p70S6K 的磷酸化水平，促进了 ILK 的表达，但对 PDK1 的表达影响较小。GDC-0941 抑制了 ILK 过表达引起的 p-AKT、p-S6 和 p-p70S6K 的增加，并促进了 ILK 敲低诱导的 p-AKT、p-S6 和 p-p70S6K 的减少。此外，OSU-T315 和 GDC-0941 的联合使用降低了 p-AKT、p-S6 和 p-p70S6K 的水平、肿瘤体积和肿瘤重量。GDC-0941 通过上调 TNF-α 水平促进了 ILK 的表达。综上所述，GDC-0941 通过上调 TNF-α 水平增加了 ILK 水平，从而影响了 AKT 的表达和乳腺癌细胞对 GDC-0941 的敏感性。
