[摘 要] 目的：探讨扁蒴藤素（PT）通过蛋白激酶B（AKT）/糖原合成酶激酶-3β（GSK-3β）信号通路对人乳腺癌细胞MCF-7多 柔比星（DOX）敏感性的影响。方法：体外培养MCF-7细胞并建立DOX耐药细胞MCF-7/DOX。MCF-7细胞设NC组、L-PT 组（2 μmol/L PT）、M-PT组（4 μmol/L PT）、H-PT组（8 μmol/L PT）、H-PT + SC79组（8 μmol/L PT + 10 μmol/L的AKT/GSK-3β信号 通路抑制剂SC79）、H-PT + LY294002组（8 μmol/L PT + 2.5 μmol/L的AKT/GSK-3β信号通路激活剂LY294002）。MCF-7/DOX细 胞设MCF-7/DOX组（未处理）、DOX组（50 nmol/L DOX）、PT + DOX组（8 μmol/L PT和50 nmol/L DOX）、PT + DOX + SC79组（8 μmol/L PT + 50 nmol/L DOX + 10 μmol/L SC79）、PT + DOX + LY294002 组（8 μmol/L PT + 50 nmol/L DOX + 2.5 μmol/L LY294002）。采用MTT法、平板克隆实验、划痕愈合实验、Transwell实验及WB法分别检测细胞增殖、集落形成数、迁移、侵袭和 AKT/GSK-3β信号通路蛋白表达。建立MCF-7细胞裸鼠移植瘤模型，观察PT对移植瘤生长、移植瘤组织中AKT/GSK--3β 信号通 路蛋白表达的影响。结果：与NC组相比，L-PT、M-PT、H-PT组MCF-7细胞增殖率、集落形成数、划痕愈合率、侵袭细胞数及 p-AKT、p-GSK-3β蛋白表达均呈PT浓度依赖性降低（均P < 0.05）；与H-PT组对比，H-PT + SC79组MCF-7细胞上述指标变化趋 势与上述相反，PT + DOX + LY294002组MCF-7细胞上述指标变化趋势与上述相同（均P < 0.05）。MCF-7/DOX组与DOX组细 胞增殖率、集落形成数、划痕愈合率、侵袭细胞数及p-AKT、p-GSK-3β蛋白表达无明显差异（均P > 0.05）；分别与MCF-7/DOX组、 DOX组对比，PT + DOX组MCF-7/DOX细胞上述指标均降低（均P < 0.05）；对比PT + DOX组，PT + DOX + SC79组上述指标均 升高，PT + DOX + LY294002组上述指标均降低（均P < 0.05）。裸鼠移植瘤实验显示，与对照组和DOX相比，PT + DOX组 移植瘤质量、体积、p-AKT、p-GSK-3β蛋白表达均降低（均P < 0.05）。结论：PT抑制乳腺癌细胞增殖、迁移和侵袭，并增强其对 DOX的化疗敏感性，其机制与抑制AKT/GSK-3β信号通路有关。

[Abstract] Objective: To investigate the effect of pristimerin (PT) on doxorubicin (DOX) sensitivity of huamn breast cancer cell MCF-7 by regulating the protein kinase B (AKT)/glycogen synthase kinase-3β (GSK-3β) signaling pathway. Methods: MCF-7 cells were cultured in vitro and used to construct a DOX resistant cell line MCF-7/DOX. MCF-7 cells were separated into the NC group, the L-PT group (2 μmol/L PT), the M-PT group (4 μmol/L PT), the H-PT group (8 μmol/L PT), the H-PT+SC79 group (8 μmol/L PT + 10 μmol/L AKT/GSK-3β signaling pathway inhibitor SC79), and the H-PT + LY294002 group (8 μmol/L PT + 2.5 μmol/L AKT/GSK-3β signaling pathway activator LY294002). MCF-7/DOX cells were separated into the MCF-7/DOX group (untreated), the DOX group (50 nμmol/L DOX), PT + DOX group (8 μmol/L PT and 50 nμmol/L DOX), the PT + DOX + SC79 group (8 μmol/L PT + 50 nμmol/L DOX + 10 μmol/L SC79), and the PT + DOX + LY294002 group (8 μmol/L PT + 50 nμmol/L DOX + 2.5 μmol/L LY294002). MTT assay, plate cloning assay, scratch assay, Transwell assay, and WB assay were applied respectively to determine cell proliferation, colony formation, migration, invasion, and AKT/GSK-3β signaling pathway protein expression in each group. Establish a MCF-7 cell xenograft model in nude mice to observe the effects of PT on tumor growth and the protein expression of the AKT/GSK-3β signaling pathway in the tumor tissues. Results: Compared with the NC group, the proliferation rate, colony formation rate, scratch healing rate, invasive cell count, and the p-AKT, p-GSK-3β protein expressions of MCF-7 cells in the L-PT group, the M-PT group, and the H-PT group all showed a PT concentration dependent decrease (all P < 0.05). Compared with the H-PT group, the trend of changes in the above indicators of MCF-7 cells in the H-PT + SC79 group was opposite to the above, while the trend of changes in the above indicators of MCF-7 cells in the PT + DOX + LY294002 group was the same (all P < 0.05). There was no significant difference in the proliferation rate, colony formation number, scratch healing rate, invasive cell count, and expressions of p-AKT and p-GSK-3β proteins between the MCF-7/DOX group and the DOX group (all P > 0.05). Compared with the MCF-7/DOX group and the DOX group, the PT + DOX group showed a decrease in the above indicators of MCF-7/DOX cells (all P < 0.05). Compared with the PT + DOX group, the above indicators in the PT + DOX + SC79 group all increased, while the above indicators in the PT + DOX + LY294002 group all decreased (all P < 0.05). Transplant tumor experiment in nude mice showed that compared with those in the control group and the DOX group, the mass and volume of transplant tumors, p-AKT and P-GSK-3β protein expressions in the PT + DOX group all decreased (all P < 0.05). Conclusion: PT can inhibit the proliferation, migration, and invasion of BC cells, and enhance their sensitivity to DOX chemotherapy., Its mechanism is related to the inhibition of the AKT/GSK-3β signaling pathway.

新疆维吾尔自治区自然科学基金（No. 2020D01C117）