目的 研究歐前胡素對人乳腺癌MCF-7/他莫昔芬（tamoxifen，TAM）的多藥耐藥逆轉(zhuǎn)作用及相關(guān)機制。方法 分別用歐前胡素、TAM、歐前胡素聯(lián)合他莫昔芬處理人乳腺癌MCF-7細(xì)胞和人乳腺癌他莫昔芬耐藥株MCF-7/TAM細(xì)胞，采用MTT法檢測MCF-7/TAM細(xì)胞耐藥性、細(xì)胞活力以及聯(lián)合用藥逆轉(zhuǎn)倍數(shù)；傷口愈合實驗、Transwell實驗、集落形成實驗檢測MCF-7/TAM細(xì)胞遷移、侵襲和集落形成能力；免疫印跡法檢測細(xì)胞多藥耐藥相關(guān)蛋白1（multidrug resistance-associated protein 1，MRP1）、多藥耐藥1（multidrug resistance 1，MDR1）、谷胱甘肽S-轉(zhuǎn)移酶π（glutathione S-transferase π ，GST-π）蛋白表達(dá)情況變化；將MCF-7/TAM細(xì)胞株接種于裸鼠皮下構(gòu)建裸鼠乳腺癌TAM耐藥移植瘤模型，分別將成功構(gòu)建移植瘤裸鼠隨機分為對照組、TAM（4.6 mg/kg）組、歐前胡素（20 mg/kg）組、TAM（4.6 mg/kg）聯(lián)合歐前胡素低劑量（10 mg/kg）組、TAM（4.6 mg/kg）聯(lián)合歐前胡素高劑量（20 mg/kg）組，ig給藥21 d，期間測量腫瘤體積并稱量體質(zhì)量；給藥結(jié)束后處死裸鼠剖取腫瘤組織稱量腫瘤質(zhì)量，采用TUNEL法檢測腫瘤細(xì)胞凋亡情況；采用免疫組化法檢測增殖標(biāo)志物Ki-67的蛋白表達(dá)情況及病理情況；采用定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)（reverse transcription quantitative polymerase chain reaction，RT-qPCR）法和免疫印跡法檢測多藥耐藥蛋白MRP1、MDR1、GST-π蛋白表達(dá)情況。結(jié)果 歐前胡素抑制MCF-7/TAM細(xì)胞增殖作用呈劑量相關(guān)性；歐前胡素聯(lián)合TAM給藥組顯著抑制細(xì)MCF-7/TAM細(xì)胞遷移、侵襲和集落形成能力（P＜0.05、0.01）、顯著下調(diào)多藥耐藥蛋白MRP1、MDR1、GST-π的蛋白表達(dá)（P＜0.05、0.01）；歐前胡素聯(lián)合TAM能夠逆轉(zhuǎn)人乳腺癌TAM耐藥的多藥耐藥性，抑制腫瘤增殖（P＜0.05）、增加腫瘤細(xì)胞凋亡（P＜0.05、0.01）、顯著下調(diào)多藥耐藥蛋白MRP1、MDR1、GST-π的蛋白表達(dá)（P＜0.05、0.01）。結(jié)論 歐前胡素可增強乳腺癌MCF-7/TAM細(xì)胞TAM的敏感性，降低MRP1、MDR1、GST-π的蛋白表達(dá)，抑制乳腺癌TAM的耐藥性。

Objective Investigate the role of imperatorin in reversing multidrug resistance in breast cancer MCF-7/tamoxifen (TAM) cells and its related mechanism. Methods Breast cancer MCF-7 cells and MCF-7/TAM cells were treated with imperatorin, TAM, and imperatorin combined with TAM in vitro, respectively. The drug resistance, cell viability and reversal ratio of MCF-7/TAM cells were detected by MTT assay. MCF-7/TAM cell migration, invasion and colony formation were evaluated by wound healing test, Transwell test and colony formation test, Western blotting was used to detect the changes in the protein expression of multidrug resistance-associated protein 1 (MRP1), multidrug resistance 1 (MDR1) and glutathione S-transferase π (GST-π). The MCF-7/TAM cell line was inoculated subcutaneously in nude mice to construct a TAM resistant xenograft model of breast cancer in nude mice, and the nude mice who successfully constructed xenografts were randomly divided into control group, TAM (4.6 mg/kg) group, imperatorin (20 mg/kg) group, TAM (4.6 mg/kg) combined with imperatorin low-dose (10 mg/kg) group, and TAM (4.6 mg/kg) combined with imperatorin high-dose (20 mg/kg) group, ig administration was performed for 21 days, during which tumor volume was measured and body weight was weighed. After the end of administration, nude mice were sacrificed to dissect tumor tissues to weigh the tumor mass, and the apoptosis of tumor cells was detected by TUNEL method. Immunohistochemistry was used to detect the expression and pathology of marker of proliferation Ki-67. Reverse transcription quantitative polymerase chain reaction，RT-qPCR and Western blotting were used to detect the protein expressions of multidrug resistance proteins MRP1, MDR1 and GST-π.Results In vitro, imperatorin inhibited the proliferation of MCF-7/TAM cells in a dose-dependent manner, and the combination of imperatorin and TAM significantly inhibited the migration, invasion and colony formation ability of fine MCF-7/TAM cells ( P < 0.05, 0.01), and significantly down-regulate the expression of multidrug resistant proteins MRP1, MDR1 and GST-π (P < 0.05, 0.01). In vivo, imperatorin combined with TAM could reverse the multidrug resistance of TAM-resistant human breast cancer, inhibit tumor proliferation (P < 0.05), increase tumor cell apoptosis (P < 0.05, 0.01), and significantly down-regulate the expression of multidrug resistant proteins MRP1, MDR1 and GST-π (P < 0.05, 0.01). Conclusion Imperatorin can enhance the TAM sensitivity of MCF-7/TAM cells, reduce the expression of MRP1, MDR1 and GST-π, and inhibit TAM resistance in breast cancer.

R285.5

國家自然科學(xué)基金項目(82060733);江西省衛(wèi)生健康委員會項目(202311141);中藥改良創(chuàng)新江西省重點實驗室(2024SSY07131);大學(xué)生校級創(chuàng)新創(chuàng)業(yè)項目(X202310412252);校級研究生創(chuàng)新專項基金(JZYC23S77,JZYC23S71);江西中醫(yī)藥大學(xué)研究生教改項目(jzyjg-2023-07)