目的 研究粉防己碱通过调控Hippo/Yes-相关蛋白同源癌蛋白（homologous oncoproteins Yes-associated protein，YAP）信号通路抗乳腺癌多重耐药的分子机制。方法 采用CCK-8法检测粉防己碱对乳腺癌MCF-7/ADR细胞增殖的影响；采用倒置显微镜、荧光显微镜观察细胞形态；采用流式细胞仪检测细胞凋亡率；采用细胞集落实验检测细胞克隆形成能力；采用Transwell实验检测细胞侵袭能力；采用Western blotting检测细胞耐药外排蛋白[P-糖蛋白（P-glycoprotein，P-gp）、乳腺癌耐药蛋白（breast cancer resistance protein，BCRP）、多药耐药相关蛋白（multidrug resistance associated protein，MRP）]，Hippo通路关键节点蛋白[YAP1、大肿瘤抑制激酶1（large tumor suppressor kinase 1，LATS1）、哺乳动物不育系20样激酶1（mammalian Sterile 20-like kinase 1，MST1）、转录共活化因子（transcriptional co-activator with PDZ-binding motif，TAZ）]和凋亡关键蛋白[半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、细胞色素C（cytochrome-C，Cyt-C）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）]表达。结果 粉防己碱对MCF-7/ADR细胞具有增殖抑制作用，半数抑制浓度（half inhibitory concentration，IC₅₀）值为14.20 μmol/L；粉防己碱能够改变细胞形态，部分细胞出现凋亡迹象；粉防己碱显著诱导MCF-7/ADR细胞发生凋亡（P＜0.01），抑制细胞的增殖分化和侵袭能力，显著下调P-gp、BCRP、MRP1、YAP1、TAZ和Bcl-2蛋白表达（P＜0.01），显著上调MST1、LATS1、Caspase-3、Cyt-C和Bax蛋白表达（P＜0.01）。结论 粉防己碱能够通过下调MCF-7/ADR细胞P-gp、BCRP和MRP1蛋白表达，抑制外排蛋白活性，增加细胞内粉防己碱含量蓄积；并通过激活Hippo信号通路，上调MST1蛋白表达，激活LATS1，进而调控下游靶点YAP1和TAZ表达，进一步上调Caspase-3、Cyt-C和Bax蛋白表达，下调Bcl-2蛋白表达，启动细胞凋亡程序，发挥抗肿瘤作用。

Objective To study the molecular mechanism of tetrandrine against multidrug resistance in breast cancer by regulating Hippo/homologous oncoproteins Yes-associated protein (YAP) signaling pathway. Methods CCK-8 method was used to detect the effect of tetrandrine on proliferation of MCF-7/ADR cells; Inverted microscopy and fluorescence microscopy were used to observe cell morphology; Flow cytometry was used to detect cell apoptosis rate; Cell colony assay was used to detect cell clone formation ability; Transwell experiment was used to detect cell invasion ability; Western blotting was used to detect expressions of resistance efflux proteins [P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and multidrug resistance associated protein (MRP)], Hippo pathway key node proteins [YAP1, large tumor suppressor kinase 1 (LATS1), mammalian Sterile 20-like kinase 1 (MST1), transcriptional co activator with PDZ binding motif (TAZ)], and apoptosis key proteins [cysteine aspartate protease-3 (Caspase-3), cytochrome C (Cyt-C), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax)]. Results Tetrandrine had a proliferative inhibitory effect on MCF-7/ADR cells, with a half inhibitory concentration (IC₅₀) value of 14.20 μmol/L; Tetrandrine could change cell morphology and some cells showed signs of apoptosis; Tetrandrine significantly induced apoptosis in MCF-7/ADR cells (P < 0.01), inhibited cell proliferation, differentiation and invasion ability, significantly down-regulated the expressions of P-gp, BCRP, MRP1, YAP1, TAZ and Bcl-2 proteins (P < 0.01), and significantly up-regulated the expressions of MST1, LATS1, Caspase-3, Cyt-C and Bax proteins (P < 0.01). Conclusion Tetrandrine can inhibit efflux protein activity and increase intracellular accumulation of tetrandrine by down-regulating the expressions of P-gp, BCRP and MRP1 proteins in MCF-7/ADR cells. And tetrandrine exerts anti-tumor effects by activating Hippo signaling pathway, upregulation of MST1 protein expression, activation of LATS1, and regulation of downstream target YAP1 and TAZ expressions, further upregulation of Caspase-3, Cyt-C, and Bax protein expressions, down-regulation of Bcl-2 protein expression, initiation of cell apoptosis program.

R285.5

中国博士后面上项目（2021MD703828）；黑龙江省自然科学基金优秀青年项目（YQ2022H002）；黑龙江省博士后资助项目（LBH-Z20172）；哈尔滨商业大学产业化项目支持计划（XL0086）