目的：探究吉马酮通过叉头盒蛋白 O3（FOXO3）-FOX 亚类 M1 转录因子（FOXM1）信号通路调控人甲状腺癌细胞 BCPAP和多柔比星（DOX）耐药BCPAP细胞（BCPAP/DOX）的增殖、迁移和化疗耐药。方法:常规培养BCPAP细胞，用BCPAP细 胞构建BCPAP/DOX细胞，用MTT法检测不同浓度吉马酮对BCPAP和BCPAP/DOX细胞增殖的影响。将BCPAP和BCPAP/DOX 细胞分为Ctrl组（空白对照）、sh-NC组（转染sh-NC质粒）、低浓度吉马酮组（0.10 mmol/L）、高浓度吉马酮组（0.15 mmol/L）、高浓 度吉马酮（0.15 mmol/L） + sh-FOXO3组（转染sh-FOXO3质粒），用转染试剂将sh-NC质粒和sh-FOXO3质粒转至相应的BCPAP 和BCPAP/DOX细胞中。用CCK-8法、划痕愈合实验和WB法分别检测各组细胞的增殖、迁移能力，以及FOXO3、FOXM1、BAX、 MMP-9和多药耐药-1（MDR-1）蛋白的表达。结果:在BCPAP和BCPAP/DOX细胞中成功地敲减了FOXO3的表达，低浓度和高 浓度吉马酮均可显著抑制BCPAP和BCPAP/DOX细胞的增殖、迁移能力，降低细胞中FOMX1、MMP-9或MDR-1（BCPAP/DOX细 胞）蛋白的表达，提升FOXO3和BAX蛋白的表达（均P < 0.05），与低浓度比较，高浓度吉马酮的作用更为显著（均P < 0.05），敲减 FOXO3后可部分逆转吉马酮对这两种细胞的作用（均P < 0.05）。结论:吉马酮可能通过FOXO3/FOXM1信号通路调控BCPAP 和BCPAP/DOX细胞的增殖和迁移能力，降低BCPAP/DOX细胞化疗耐药性。

Objective: To explore how germacrone regulates the proliferation, migration and chemoresistance of human thyroid cancer BCPAP cells and doxorubicin (DOX) resistant BCPAP cells (BCPAP/DOX) through the forkhead box O3 (FOXO3)-FOX subclass M1 transcription factor (FOXM1) signaling pathway. Methods: BCPAP cells were cultured routinely and used to construct BCPAP/DOX cells. MTT method was applied to detect the effects of different concentrations of germacrone on the proliferation of BCPAP and BCPAP/DOX cells. BCPAP and BCPAP/DOX cells were divided into control group (Ctrl), negative control group (NC, transfected with sh-NC plasmid), low concentration germacrone group (0.10 mmol/L), high concentration germacrone group (0.15 mmol/L), high concentration germacrone (0.15 mmol/L) + sh-FOXO3 group (transfected with sh-FOXO3 plasmid). The sh-NC plasmid and sh-FOXO3 plasmid were transfected into the corresponding BCPAP and BCPAP/DOX cells with transfection reagents. The proliferation and migration of the cells were detected using CCK-8 assay and scratching healing assay, and the expression of FOXO3, FOXM1, BAX, MMP-9 and multi-drug resistant-1 (MDR-1) was detected using WB assay. Results: The expression of FOXO3 was successfully knocked down in BCPAP and BCPAP/DOX cells. Both low and high concentrations of germacrone could significantly inhibit the proliferation and migration of BCPAP and BCPAP/DOX cells, reduce the protein expression of FOMX1, MMP-9 or MDR-1 (in BCPAP/ DOX cells), and increase the protein expression of FOXO3 and BAX (all P < 0.05). Notably, the effects were more significant with high concentration germacrone compared to that of low concentration (all P < 0.05). Knockdown of FOXO3 partially reversed the effects of germacrone on these cells (all P < 0.05). Conclusion: Germacrone may regulate the proliferation and migration of BCPAP and BCPAP/DOX cells and reduce chemotherapy resistance of BCPAP/DOX cells through the FOXO3/FOXM1 signaling pathway.

河北省医学科学研究课题计划项目（No.20220384）