ATF_(24)多肽修饰的β-榄香烯脂质体联合顺铂对膀胱癌5637细胞的协同增效作用  

Synergistic Effect of ATF_(24) Peptide-Modifiedβ-elemene Liposomes Combined with Cisplatin on Bladder Cancer Cell Line 5637

作　　者：翟秉涛 田欢 孙静[1] 史亚军[1] 张小飞[1] 邹俊波[1] 程江雪[1] 郭东艳[1] ZHAI Bing-tao;TIAN Huan;SUN Jing;SHI Ya-jun;ZHANG Xiao-fei;ZOU Jun-bo;CHENG Jiang-xue;GUO Dong-yan(State Key Laboratory of Research and Development of Characteristic Qin Medicine Resources(Cultivation),Shaanxi Key Laboratory of Chinese Medicine Fundamentals and New Drugs Research,Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Shaanxi University of Chinese Medicine,Xi′an 712046,China;Xi′an Traditional Chinese Medicine Hospital,Xi′an 710001,China)

机构地区：[1]陕西中医药大学,秦药特色资源研究与开发国家重点实验室(培育),陕西省中药基础与新药研究重点实验室,西安712046 [2]西安市中医医院,西安710001

出　　处：《中国药学杂志》2023年第6期506-515,共10页Chinese Pharmaceutical Journal

基　　金：陕西省自然科学基础研究计划项目资助(2022JQ-917,2022JQ-932);陕西省教育厅重点科研计划项目资助(21JS009);西安市卫生健康委员会科研项目资助(2022yb01);陕西中医药大学学科创新团队项目资助(2019-YL11);陕西中医药大学校级科研课题资助(2021GP04);国家自然科学基金资助项目(82204935)。

摘　　要：目的研究ATF_(24)多肽修饰的β-榄香烯脂质体(ATF_(24)-PEG-Lipo-β-E)联合顺铂(DDP)对膀胱癌5637细胞的协同增效作用。方法通过免疫印迹法(Western blot)测定膀胱癌5637细胞中尿激酶纤溶酶原受体(uPAR)的表达;利用激光扫描共聚焦显微镜和高效液相色谱法(HPLC)研究ATF_(24)多肽修饰的脂质体的摄取情况;通过Transwell实验研究脂质体对5637细胞迁移和侵袭的影响;通过细胞计数-8(CCK-8)法、集落形成实验以及流式细胞术研究ATF_(24)-PEG-Lipo-β-E联合DDP对5637细胞增殖、细胞凋亡和细胞周期的影响;通过Western blot测定凋亡相关蛋白剪切的聚腺苷酸二磷酸核糖转移酶(Cleaved PARP)和蛋白B细胞淋巴瘤-2(Bcl-2),以及周期相关蛋白细胞周期蛋白B1(Cyclin B1)和细胞分裂周期蛋白25C(Cdc25C)的表达水平。结果与RT4细胞和KU-19-19细胞相比,5637细胞高表达uPAR蛋白;细胞摄取定性和定量结果显示,ATF_(24)多肽介导的脂质体相比于长循环脂质体对5637细胞具有更好的靶向性;Transwell实验结果显示,ATF_(24)-PEG-Lipo-β-E能够通过靶向uPAR降低细胞迁移和侵袭;CCK-8实验和集落形成实验表明,ATF_(24)-PEG-Lipo-β-E和DDP联用能够明显抑制5637细胞增殖;两药联用还能够通过上调Cleaved PARP蛋白的表达,下调Bcl-2蛋白的表达诱导5637细胞凋亡,以及通过下调Cyclin B1和Cdc25C蛋白的表达进一步阻滞细胞周期于G2/M期。结论ATF_(24)-PEG-Lipo-β-E和DDP联用对膀胱癌5637细胞具有协同增效作用。OBJECTIVE To study the synergistic effect of ATF_(24)peptide-modifiedβ-elemene liposomes(ATF_(24)-PEG-Lipo-β-E)combined with cisplatin(DDP)on bladder cancer cell line 5637.METHODS The expression of urokinase plasminogen receptor(uPAR)in bladder cancer 5637 cells was determined by Western blot;the cellular uptake of ATF_(24)polypeptide modified liposomes was studied by confocal laser scanning microscopy and high performance liquid chromatography(HPLC);the effect of liposomes on cell migration and invasion was studied by transwell assay;the effect of ATF_(24)-PEG-Lipo-β-E combined with DDP on cell proliferation,cell apoptosis and cell cycle were studied by cell counting kit-8(CCK-8)assay,colony formation assay and flow cytometry;the expressions of apoptosis-related proteins cleaved poly ADP-ribose polymerase(Cleaved PARP)and proteins B lymphocytoma-2(Bcl-2),and the expressions of cycle-related proteins Cyclin B1 and cell division cyclin 25C(Cdc25C)were determined by Western blot.RESULTS Compared with RT4 cells and KU-19-19 cells,uPAR had higher expression in 5637 cells;the qualitative and quantitative tests of cellular uptake showed that the ATF_(24)peptide-modified liposomes had better targeting on 5637 cells than long-circulating liposomes;the transwell assay showed that ATF_(24)-PEG-Lipo-β-E could reduce cell migration and invasion by targeting uPAR;CCK-8 assay and colony formation assay showed that the combination of ATF_(24)-PEG-Lipo-β-E and DDP could significantly inhibit the proliferation of 5637 cells;the combination of the two drugs could also induce cell apoptosis by upregulating the expression of cleaved PARP and downregulating the expression of Bcl-2,and block the cell cycle at G2/M phase by downregulating the expression of Cyclin B1 and Cdc25C.CONCLUSIONS ATF_(24)-PEG-Lipo-β-E has a synergistic effect with DDP on bladder cancer cell line 5637.

关 键 词：Β-榄香烯 ATF_(24)多肽 脂质体 尿激酶纤溶酶原受体 顺铂 膀胱癌 

分 类 号：R966[医药卫生—药理学]