Department of Gynecology, Fourth Hospital of Hebei Medical University, No.12, Jiankang Road, Shijiazhuang, 050011, Hebei, China.
Cancer Chemother Pharmacol. 2021 Aug;88(2):323-334. doi: 10.1007/s00280-021-04291-5. Epub 2021 May 12.
The present study aims to examine the effects of apatinib combined with autophagy inhibitor 3-Methyladenine (3-MA) on the proliferation and apoptosis of human uterine sarcoma in FU-MMT-1 and MES-SA cells and its tumor inhibition effect in xenograft model of uterine sarcoma.
Different concentrations of 3-MA and apatinib were used to treat the uterine sarcoma cell lines (MES-SA and FU-MMT-1 cells). The cell viability was detected by CCK8 method. Flow cytometry was used to detect the apoptosis and cell cycle. Wound closure assay and Transwell assay were performed to measure the migration ability of cells. Western blot was used to determine the apoptosis proteins and autophagy proteins. A nude mice sarcoma xenograft model was established and treated with apatinib alone, 3-MA alone, or combined incubation of them. Tumor size of xenograft and the mice survival rate were measured.
Combination of 3-MA and apatinib significantly inhibited the proliferation and migration ability, but increased the apoptosis rate of uterine sarcoma cells compared to apatinib. The combination of 3-MA and apatinib significantly limited the tumor size of xenograft and increased the survival rate of mice compared to apatinib alone. Apatinib inhibited the PI3K/Akt/mTOR pathway, while 3-MA and the combination of 3-MA and apatinib significantly activated the PI3K/Akt/mTOR pathway and inhibited autophagy. Combination of 3-MA and apatinib increased apoptosis compared to apatinib alone. The expression of VEGFR-2 was not impacted by 3-MA.
Combination of apatinib and autophagy inhibitor 3-MA significantly inhibited the growth and migration of uterine sarcoma cells and xenograft. Autophagy inhibition may increase the antitumor effect of apatinib via the PI3K/Akt/mTOR pathway.
本研究旨在探讨阿帕替尼联合自噬抑制剂 3-甲基腺嘌呤（3-MA）对人子宫肉瘤 FU-MMT-1 和 MES-SA 细胞增殖和凋亡的影响及其在子宫肉瘤异种移植模型中的抑瘤作用。
用不同浓度的 3-MA 和阿帕替尼处理子宫肉瘤细胞系（MES-SA 和 FU-MMT-1 细胞）。用 CCK8 法检测细胞活力。用流式细胞术检测细胞凋亡和细胞周期。用划痕闭合实验和 Transwell 实验检测细胞迁移能力。用 Western blot 检测凋亡蛋白和自噬蛋白。建立裸鼠肉瘤异种移植模型，并分别用阿帕替尼、3-MA 或两者联合处理。测量肿瘤体积和小鼠存活率。
与阿帕替尼相比，3-MA 与阿帕替尼联合应用显著抑制了子宫肉瘤细胞的增殖和迁移能力，但增加了其凋亡率。与阿帕替尼单独治疗相比，3-MA 与阿帕替尼联合应用显著限制了异种移植瘤的大小并提高了小鼠的存活率。阿帕替尼抑制了 PI3K/Akt/mTOR 通路，而 3-MA 和 3-MA 与阿帕替尼的联合应用显著激活了 PI3K/Akt/mTOR 通路并抑制了自噬。与阿帕替尼单独治疗相比，3-MA 与阿帕替尼联合应用增加了凋亡。3-MA 对 VEGFR-2 的表达没有影响。
阿帕替尼与自噬抑制剂 3-MA 的联合应用显著抑制了子宫肉瘤细胞和异种移植瘤的生长和迁移。自噬抑制可能通过 PI3K/Akt/mTOR 通路增加阿帕替尼的抗肿瘤作用。
