摘要
目的探讨氯硝柳胺抑制紫杉醇耐药性三阴性乳腺癌(TNBC)的效果,并分析可能的作用机制。方法培养紫杉醇耐药性TNBC细胞株MDA-MB-231,对照组使用紫杉醇(1μmol/L)处理,氯硝柳胺组使用氯硝柳胺(3μmol/L)处理,联合用药组使用磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)通路抑制剂LY294002(1μmol/L)+氯硝柳胺(3μmol/L)处理。采用MTT试验检测各组细胞增殖能力;细胞凋亡检测试剂盒检测细胞凋亡情况;Transwell小室试验检测细胞迁移能力;Western blot法检测细胞p-PI3K、PI3K、p-Akt、Akt、裂解的半胱氨酸蛋白酶-3(Caspase-3)、P-糖蛋白(P-gP)表达水平。将耐药性MDA-MB-231细胞接种至雄性SD小鼠体内,建立小鼠异种移植肿瘤模型并分为3组,分别腹腔注射紫杉醇、氯硝柳胺、氯硝柳胺+LY294002,每日测量肿瘤大小并计算肿瘤体积;5周后采用免疫组织化学法检测小鼠肿瘤组织Ki-67表达情况。结果与对照组相比,氯硝柳胺组、联合用药组细胞增殖能力、迁移能力均下降,细胞凋亡增多;细胞p-PI3K/PI3K、p-Akt/Akt蛋白表达水平均下降,裂解的Cas-pase3蛋白表达水平均上升,P-gp蛋白表达水平均下降,差异均有统计学意义(均P<0.05)。氯硝柳胺组、联合用药组小鼠5周后与对照组比较,移植肿瘤体积均缩小(均P<0.05),移植肿瘤组织Ki-67表达均下调(均P<0.05)。结论氯硝柳胺通过PI3K/Akt通路抑制紫杉醇耐药性TNBC细胞的体内外增殖,并诱导细胞凋亡。
Objective To investigate the effect of niclosamide on proliferation and migration of paclitaxel-resistant triple-negative breast cancer(TNBC)cells and related mechanism.Methods The paclitaxel-resistant TNBC MDA-MB 231 cells were treated with 1μmol/L paclitaxel,3μmol/L niclosamide(niclosamide group)or 1μmol/L LY294002(PI3K/Akt pathway inhibitor)and 3μmol/L niclosamide(niclosamide+LY294002 group),respectively.MTT assay was used to detect cell proliferation ability;apoptosis detection kit was used to detect cell apoptosis;Transwell chamber test was used to detect cell migration ability;Western blot method was used to detect expression levels of p-PI3K,PI3K,p-Akt,Akt,cleaved cysteine,cleaved Caspase 3 and P-glycoprotein(P-gp)in cells.Male SD mice were inoculated with 3 groups of drug resistant MDA-MB-231 cells to establish a mouse xenograft tumor model,respectively.The tumor size was measured with a vernier caliper every day,and the tumor volume was calculated.After 5 weeks of intervention,the mice were sacrificed by cervical dislocation and the tumor was isolated.The expression of Ki 67 in the tumor tissue of the mice was detected by immunohistochemistry.Results Compared with the control group,the cell proliferation and migration ability of niclosamide group and the niclosamide+LY294002 group were decreased,and the cell apoptosis was increased;the protein expression levels of p-PI3K/PI3K and p-Akt/Akt in cells were decreased,the protein expression level of cleaved Caspase 3 was increased,and the protein expression level of P-gp was decreased(all P<0.05);the volume of transplanted tumors in mice was reduced(all P<0.05),and the expression of Ki 67 in transplanted tumor tissues was down-regulated(all P<0.05).Conclusion Niclosamide inhibits proliferation,migration and induces apoptosis in vitro and in vivo of paclitaxel-resistant triple-negative breast cancer cells via PI3K/Akt pathway.
作者
叶玉萍
林崇峰
郑楠
YE Yuping;LIN Chongfeng;ZHENG Nan(Department of Clinical Pharmacy,Taishun County People's Hospital,Taishun 325500,China)
出处
《浙江医学》
CAS
2022年第19期2033-2038,I0004,共7页
Zhejiang Medical Journal
基金
泰顺县科技计划项目(2021TSXM0055)。
关键词
P糖蛋白
氯硝柳胺
三阴性乳腺癌
紫杉醇
增殖
凋亡
P-glycoprotein
Niclosamide
Triple negative breast cancer
Paclitaxel
Proliferation
Apoptosis
