疏肝化痰散结方与紫杉醇抗乳腺癌MCF-7/PTX细胞的协同增效作用及机制研究  

Synergistic Effect and Mechanism of Shugan Huatan Sanjie Recipe and Paclitaxel Against Breast Cancer MCF-7/PTX Cells

作　　者：张立柱 韩长辉 范焕芳[1] 张洋 ZHANG Lizhu;HAN Changhui;FAN Huanfang;ZHANG Yang(Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China;Nangong People’s Hospital,Xingtai 055750,China)

机构地区：[1]河北省中医院,石家庄050011 [2]南宫市人民医院,邢台055750

出　　处：《世界科学技术-中医药现代化》2024年第9期2476-2482,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：河北省中医药管理局科研计划项目(2021024):疏肝解郁化痰中药联合化疗对乳腺癌凝血功能的作用及临床疗效评价,负责人:张立柱。

摘　　要：目的探讨疏肝化痰散结方(SHSF)与紫杉醇抗乳腺癌MCF-7/PTX细胞的协同增效作用及机制。方法采用低浓度加量持续诱导法建立乳腺癌PTX耐药细胞株MCF-7/PTX,MTT法检测不同浓度PTX对MCF-7和MCF-7/PTX细胞及SHSF含药血清对MCF-7/PTX细胞增殖活性的影响,并根据IC50值计算耐药指数(RI)和逆转倍数。将MCF-7/PTX细胞分为空白对照组、PTX组(45 nmol·L^(-1)PTX)、SHSF组(7.5%SHSF含药血清)和PTX+SHSF组(45 nmol·L^(-1)PTX+7.5%SHSF含药血清),药物作用24 h,流式细胞术检测各组细胞凋亡水平;Western blot检测各组细胞中凋亡蛋白Bax、Bcl-2和Akt/mTOR通路相关蛋白p-Akt(ser473)、Akt、p-mTOR(Ser2448)、mTOR等蛋白表达水平。结果成功建立乳腺癌PTX耐药细胞株MCF-7/PTX,RI值为6.70。随着SHSF含药血清的浓度增加,MCF-7/PTX细胞增殖活性呈浓度依赖性降低;7.5%SHSF含药血清对MCF-7/PTX细胞的耐药性逆转倍数为3.48。与空白对照组比较,PTX组和SHSF组MCF-7/PTX细胞凋亡水平显著增加(P<0.01),Bax蛋白表达水平显著上调(P<0.01),Bcl-2、p-Akt/Akt和p-mTOR/mTOR蛋白表达水平显著下调(P<0.05);与PTX组相比,PTX+SHSF组细胞凋亡水平显著增加(P<0.01),Bax蛋白表达水平显著上调(P<0.01),Bcl-2、p-Akt和p-mTOR蛋白表达水平显著下调(P<0.01)。结论疏肝化痰散结方通过抑制Akt/mTOR通路促进细胞凋亡,从而发挥与紫杉醇抗乳腺癌MCF-7/PTX细胞的协同增效作用。Objective To investigate the synergistic effect of Shugan Huatan Sanjie prescription(SHSF)and paclitaxel on MCF-7/PTX cells of breast cancer and its mechanism.Methods The PTX-resistant breast cancer cell line MCF-7/PTX was established by continuous induction of low concentration.The effects of different concentrations of PTX on MCF-7 and MCF-7/PTX cells.Then the proliferation of MCF-7/PTX cells by SHSF containing serum were detected by MTT assay,and the drug resistance index(RI)and reversion times were calculated according to the IC50 value.MCF-7/PTX cells were divided into blank control group,PTX group(45 nmol·L^(-1) PTX),SHSF group(7.5%SHSF drugcontaining serum)and PTX+SHSF group(45 nmol·L^(-1) PTX+7.5%SHSF drug-containing serum).The cells were treated with drugs for 24 h.The apoptosis level of each group was detected by flow cytometry.The expression levels of apoptotic proteins Bax,Bcl-2,p-Akt(ser473),Akt,p-mTOR(Ser2448)and mTOR were detected by Western blot.Results The PTX-resistant cell line MCF-7/PTX was established successfully,and the RI value was 6.70.The proliferative activity of MCF-7/PTX cells decreased in a concentration-dependent manner with the increase of SHSF drug-containing serum concentration.And the resistance reversal ratio of 7.5%SHSF serum to MCF-7/PTX cells was 3.48.Compared with blank control group,the apoptosis levels of MCF-7/PTX cells in PTX group and SHSF group were significantly increased(P<0.01),the protein expression level of Bax was significantly up-regulated(P<0.01),and the protein expression levels of Bcl-2,p-Akt/Akt and p-mTOR/mTOR were significantly down-regulated(P<0.05);Compared with PTX group,the apoptosis level in PTX+SHSF group was significantly increased(P<0.01),the expression level of Bax protein was significantly up-regulated(P<0.01),and the expression levels of Bcl-2,p-Akt and p-mTOR protein were significantly down-regulated(P<0.01).Conclusion Shugan Huatan Sanjie Recipe promotes apoptosis by inhibiting Akt/mTOR pathway,and thus plays a synergistic effect with pacli

关 键 词：疏肝化痰散 乳腺癌 紫杉醇耐药 细胞凋亡 Akt/mTOR通路 

分 类 号：R285.5[医药卫生—中药学]