Hsp90靶向抑制多肽P7与多西他赛联用对肺腺癌细胞系A549的协同抗肿瘤作用

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (5): 639-643.

Hsp90靶向抑制多肽P7与多西他赛联用对肺腺癌细胞系A549的协同抗肿瘤作用

吴友明, 刘娜斯, 曹明月, 黄薇, 刘雁勇, 杨楠^*

中国医学科学院基础医学研究所北京协和医学院基础学院药理学系, 北京 100005

收稿日期:2020-02-13 修回日期:2020-03-18 出版日期:2020-05-05 发布日期:2020-04-30
通讯作者: *yangnan@ibms.pumc.edu.cn
基金资助:
国家科技重大专项(2019ZX0930117);国家自然科学基金(81972688);北京市自然科学基金(7172134,7192128);中国医学科学院医学与健康科技创新工程(2016-I2M-3-004);协和青年科研基金与中央高校基本科研业务费专项基金(3332015113,2017350002)

Synergistic antitumor effect of Hsp90 targeting-inhibitory peptide P7 combined with docetaxel in lung adenocarcinoma cell line A549

WU You-ming, LIU Na-si, CAO Ming-yue, HUANG Wei, LIU Yan-yong, YANG Nan^*

Department of Pharmacology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China

Received:2020-02-13 Revised:2020-03-18 Online:2020-05-05 Published:2020-04-30
Contact: *yangnan@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的研究热休克蛋白90(Hsp90)双功能靶向抑制多肽LPLTPLP(P7)与临床化疗药物多西他赛(DTX)联用对肺腺癌细胞系A549的影响。方法以DTX与P7单独或联合使用对A549细胞进行处理,48 h后,用CCK-8法评价细胞活性;通过流式细胞计量术检测DTX与P7单独或联合使用后的A549细胞凋亡率;用蛋白免疫印迹技术检测DTX与P7单独或联合使用对A549细胞凋亡以及耐药相关蛋白表达的影响。结果 DTX与P7对细胞存活具有协同抑制作用;联合用药组细胞凋亡率为25.8%,显著高于DTX组(P＜0.05);联合用药组凋亡相关蛋白Bcl-2、PARP显著下调(P＜0.05);cleaved-PARP显著上调(P＜0.05);耐药蛋白主要穹窿蛋白(MVP)显著下调(P＜0.05)。结论 DTX协同靶向性多肽P7能显著提高A549细胞的凋亡率,其机制可能与降低凋亡抑制蛋白Bcl-2表达水平、失活DNA 修复酶PARP及降低耐药蛋白MVP表达有关。

关键词: 肺腺癌, 协同作用, 细胞凋亡, 细胞耐药, 多肽

Abstract: Objective To investigate the effects of combination of Hsp90 bifunctional targeting-inhibitory peptide LPLTPLP (P7) with chemotherapy drug docetaxel(DTX) on lung adenocarcinoma cell line A549. Methods DTX and peptide P7 alone or combined were incubated with A549 cell for 48 hours, thereafter the effect of the drug on cell viability was evaluated using the CCK-8 method. Flow cytometry was carried out to quantify apoptotic cells. The expression of apoptotic and drug resistant protein were assessed by Western blot analysis. Results DTX and P7 exhibited a synergistic effect on cell survival. The apoptosis rate of the combined drug group was 25.8%, which was significantly higher than that of DTX group(P＜0.05). Apoptosis-related proteins Bcl-2 and PARP were significantly down-regulated(P＜0.05), and cleaved-PARP protein was significantly up-regulated(P＜0.05). Meanwhile, drug-resistant protein major vault protein(MVP) was significantly down-regulated(P＜0.05). Conclusions DTX ombined with targeted peptide P7 can significantly increase the apoptosis rate of A549 cells, which may be related to expression level of Bcl-2, inactivation of DNA repair enzyme PARP and down-regulation of multidrug-resistant protein MVP.

Key words: lung adenocarcinoma, synergy, apoptosis, drug-resistance, heptapeptide

中图分类号:

R963

吴友明, 刘娜斯, 曹明月, 黄薇, 刘雁勇, 杨楠. Hsp90靶向抑制多肽P7与多西他赛联用对肺腺癌细胞系A549的协同抗肿瘤作用[J]. 基础医学与临床, 2020, 40(5): 639-643.

WU You-ming, LIU Na-si, CAO Ming-yue, HUANG Wei, LIU Yan-yong, YANG Nan. Synergistic antitumor effect of Hsp90 targeting-inhibitory peptide P7 combined with docetaxel in lung adenocarcinoma cell line A549[J]. Basic & Clinical Medicine, 2020, 40(5): 639-643.

参考文献

[1]Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68:394-424.
[2]Hayashi Y, Kuriyama H, Umezu H, et al. Class III beta-tubulin expression in tumor cells is correlated with resistance to docetaxel in patients with completely resected non-small-cell lung cancer[J]. Intern Med, 2009,48:203-208.
[3]Jemal A, Siegel R, Ward E, et al.Cancer statistics, 2008[J]. CA Cancer J Clin, 2008,58:71-96.
[4]Wang TH, Wang HS, Soong YK. Paclitaxel-induced cell death: where the cell cycle and apoptosis come together[J]. Cancer, 2000,88:2619-2628.
[5]Whitesell L, and Lindquist SL. HSP90 and the chaperoning of cancer[J]. Nat Rev Cancer, 2005,5:761-772.
[6]Banerji U, Sain N, Sharp SY, et al. An in vitro and in vivo study of the combination of the heat shock protein inhibitor 17-allylamino-17-demethoxygeldanamycin and carboplatin in human ovarian cancer models[J]. Cancer Chemother Pharmacol, 2008,62:769-778.
[7]Iyer G, Morris MJ, Rathkopf D, et al. A phase I trial of docetaxel and pulse-dose 17-allylamino-17-demethoxygeldanamycin in adult patients with solid tumors[J]. Cancer Chemother Pharmacol, 2012,69:1089-1097.
[8]Jiang Y, Yang N, Zhang H, et al.Enhanced in vivo antitumor efficacy of dual-functional peptide-modified doce-taxel nanoparticles through tumor targeting and Hsp90 inhibition[J]. J Control Release, 2016,221:26-36.
[9]Huang W, Yuan X, Sun T, et al. Proteasome inhibitor YSY01A abrogates constitutive STAT3 signaling via down-regulation of Gp130 and JAK2 in human A549 lung cancer cells[J]. Front Pharmacol, 2017,8:1-10.
[10]Tang L, Liu J, Zhu L, et al.Curcumin inhibits growth of human NCI-H292 lung squamous cell carcinoma cells by increasing FOXA2 expression[J]. Front Pharmacol, 2018,9:1-9.
[11]Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors[J]. Adv Enzyme Regul, 1984,22:27-55.
[12]Mahalingam D, Swords R, Carew JS, et al. Targeting HSP90 for cancer therapy[J]. Br J Cancer, 2009,100:1523-1529.
[13]Solit DB, Chiosis G. Development and application of Hsp90 inhibitors[J]. Drug Discov Today, 2008,13:38-43.
[14]Pacey S, Wilson RH, Walton M, et al. A phase I study of the heat shock protein 90 inhibitor alvespimycin (17-DMAG) given intravenously to patients with advanced solid tumors[J]. Clin Cancer Res, 2011,17:1561-1570.
[15]Scheffer GL, Wijngaard PL, Flens MJ, et al. The drug resistance-related protein LRP is the human major vault protein[J]. Nat Med, 1995,1:578-582.