Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells

ZUO Ling; LI Yutong; XIANG Lingbao; CHEN Ruiqi; XIONG Hong; LIU Hongwei

doi:10.13422/j.cnki.syfjx.20240417

您当前的位置：

首页 >

文章列表页 >

Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells

更新时间：2024-07-31

- Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 30, Issue 17, Pages: 87-94(2024)
- 作者机构：
  
  1.广东医科大学附属第二医院，广东湛江 524003
  2.广东医科大学附属医院，广东湛江 524001
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20240417
  CLC： R242;R2-031;R285.5;R737.14;R22;R33
- Published：05 September 2024，
  
  Published Online：09 March 2024，
  
  Received：22 December 2023，
- 稿件说明：
扫描看全文
左玲,李煜桐,向玲宝等.计算机辅助药物设计和实验验证探讨柴胡皂苷D诱导膀胱癌细胞凋亡的分子机制[J].中国实验方剂学杂志,2024,30(17):87-94.

ZUO Ling,LI Yutong,XIANG Lingbao,et al.Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(17):87-94.
左玲,李煜桐,向玲宝等.计算机辅助药物设计和实验验证探讨柴胡皂苷D诱导膀胱癌细胞凋亡的分子机制[J].中国实验方剂学杂志,2024,30(17):87-94. DOI： 10.13422/j.cnki.syfjx.20240417.

ZUO Ling,LI Yutong,XIANG Lingbao,et al.Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(17):87-94. DOI： 10.13422/j.cnki.syfjx.20240417.

摘要

目的

通过计算机辅助药物设计及实验验证探讨柴胡皂苷D（SSD）靶向信号转导和转录激活因子3（STAT3）诱导膀胱癌细胞凋亡的作用。

方法

通过贝叶斯分类器模型探究SSD的可药性和生物毒性，通过中药系统药理学数据库与分析平台（TCMSP）检索柴胡有效

成分SSD的相关信息，通过有机小分子生物活性数据（PubChem）、TCMSP、中药分子机理的生物信息学分析工具（BATMAN-TCM）、Coremine、中医药百科全书（ETCM）和SwissTargetPrediction数据库预测SSD的作用靶点，利用GeneCards、药物靶标数据库（TTD）和在线人类孟德尔遗传（OMIM）数据库预测膀胱癌的潜在治疗靶点，将两者取交集进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。通过分子对接探究SSD与靶点蛋白结合能力和结构稳定性。使用Cytoscape 3.9.1软件构建STAT3-药物调控网络和STAT3-凋亡调控网络。分别用0、5、10、15 μmol·L

-1

的SSD处理UM-UC-3细胞24 h，采用流式细胞术检测SSD对膀胱癌细胞凋亡的影响，通过蛋白免疫印迹法（Western blot）检测B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、Bcl-2关联死亡启动子（Bad）、STAT3和磷酸化（p）-STAT3蛋白表达水平。

结果

贝叶斯分类器模型和分子对接表明，SSD具有较低生物毒性且能与靶点蛋白STAT3良好结合形成稳定蛋白-配体复合物。膀胱癌与SSD之间共有282个交集靶点，其中STAT3为最核心的靶点。GO富集分析结果显示，潜在核心治疗靶点涉及生物学过程3 036个、细胞组分82个、分子功能171个。KEGG富集分析结果显示，潜在核心靶点主要与C型凝集素受体信号通路、Toll样受体信号传导途径和细胞凋亡通路有关。STAT3-药物和STAT3-凋亡调控互作网络显示，29种药物与STAT3存在相互作用关系，27个凋亡相关基因与STAT3具有强相关性。流式细胞术显示，与空白组比较，细胞凋亡率随着SSD浓度的增加而增大（

0.05）。Western blot结果显示，与空白组比较，SSD呈浓度依赖性下调p-STAT3、Bcl-2蛋白水平，明显上调Bax、Bad蛋白水平（

0.05）。

结论

SSD具有良好的可药性和较低的生物学毒性，其可能通过靶向调控STAT3促进膀胱癌细胞凋亡。

Abstract

Objective

To explore the role of saikosaponin D （SSD） targeting signal transducer and activator of transcription 3 （STAT3） in inducing apoptosis of bladder cancer cells by computer-aided drug design and experimental verification.

Method

The druggability and biotoxicity of SSD were explored by Bayesian classifier modeling. The information about SSD， the active ingredient of Bupleuri Radix， was searched against the Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform （TCMSP）. The targets of SSD were predicted by PubChem， TCMSP， a Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）， Coremine， an Encyclopedia of Traditional Chinese Medicine （ETCM）， and SwissTargetPrediction. GeneCards， Therapeutic Target Database （TTD）， and Online Mendelian Inheritance in Man （OMIM） were employed to predict the potential therapeutic

targets of bladder cancer. Then， the common targets shared by SSD and bladder cancer were selected for Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses. Molecular docking was adopted to explore the binding affinity and structural stability of SSD with target proteins. Cytoscape 3.9.1 was used to construct the STAT3-drug regulatory network and STAT3-apoptosis regulatory network. UM-UC-3 cells were treated with 0， 5， 10， 15 μmol·L

-1

SSD for 24 h. Then， flow cytometry was used to detect the apoptosis of bladder cancer cells， and Western blot was employed to determine the protein levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， Bcl-2-associated death promoter （Bad）， STAT3， and phosphorylation （p）-STAT3.

Result

Bayesian classifier modeling and molecular docking showed that SSD had low biotoxicity and bound well to the target protein STAT3 to form a stable protein-ligand complex. There were 282 common targets between bladder cancer and SSD， among which STAT3 was the most central target. The GO enrichment analysis showed that the potential core therapeutic targets involved 3 036 biological processes， 82 cellular components， and 171 molecular functions. The KEGG enrichment analysis showed that the potential core targets were mainly related to the C-type lectin receptor signaling pathway， Toll-like receptor signaling pathway， and cell apoptosis pathway. The STAT3-drug regulatory network and STAT3-apoptosis regulatory network showed that 29 drugs interacted with STAT3， and 27 apoptosis-related genes had a strong correlation with STAT3. Flow cytometry showed that the apoptosis rate increased with the increase in SSD concentration （

0.05）. Western blotting results showed that SSD down-regulated the protein levels of p-STAT3 and Bcl-2 and up-regulated the protein levels of Bax and Bad in a concentration-dependent manner （

0.05）.

Conclusion

SSD has good druggability and low biotoxicity. It may promote the apoptosis of bladder cancer cells by targeting STAT3.

关键词

柴胡皂苷D网络药理学膀胱癌凋亡信号转导和转录激活因子3（STAT3）

Keywords

saikosaponin Dnetwork pharmacologybladder cancerapoptosissignal transducer and activator of transcription 3 （STAT3）

references

SUNG H，FERLAY J，SIEGEL R L，et al.Global cancer statistics 2020：GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］.CA Cancer J Clin，2021，71（3）：209-249.

LENIS A T，LEC P M，CHAMIE K，et al.Bladder cancer：A review［J］.JAMA，2020，324（19）：1980-1991.

BABJUK M，BURGER M，CAPOUN O，et al.European association of urology guidelines on non-muscle-invasive bladder cancer （Ta， T1， and carcinoma in situ）［J］.Eur Urol，2022，81（1）：75-94.

CATHOMAS R，LORCH A，BRUINS H M，et al.The 2021 updated european association of urology guidelines on metastatic urothelial carcinoma［J］.Eur Urol，2022，81（1）：95-103.

KONALA V M，ADAPA S，ARONOW W S.Immunotherapy in bladder cancer［J］.Am J Ther，2022，29（3）：e334-e337.

HU J，CHEN J，OU Z，et al.Neoadjuvant immunotherapy，chemotherapy，and combination therapy in muscle-invasive bladder cancer：A multi-center real-world retrospective study［J］.Cell Rep Med，2022，3（11）：100785.

LUO H，VONG C T，CHEN H，et al.Naturally occurring anti-cancer compounds：Shining from Chinese herbal medicine［J］.Chin Med，2019，14：48.

ZHANG X，QIU H，LI C，et al.The positive role of traditional Chinese medicine as an adjunctive therapy for cancer［J］.Biosci Trends，2021，15（5）：283-298.

WANG X，LI J，CHEN R，et al.Active ingredients from Chinese medicine for combination cancer therapy［J］.Int J Biol Sci，2023，19（11）：3499-3525.

LI D，ZUO M，HU X.Global trends in research of treatment on bladder cancer with chinese medicine monomer from 2000 to 2021： A bibliometric analysis［J］.J Oncol，2022，2022：3382360.

QI H，PAN Y，CHEN L，et al.Effect of chinese medicine on the biological behavior and magnetic resonance imaging of bladder cancer［J］.Evid Based Complement Alternat Med，2021，2021：1124526.

胡锦丹，王宇.中医药治疗膀胱癌临床研究进展及机制概述［J］.中医药学报，2022，50（1）：91-95.

杨明，叶和松，秦远，等.彭培初自拟方联合吉西他滨治疗非肌层浸润性膀胱癌术后的临床观察［J］.中成药，2019，41（5）：1188-1190.

孙凯廷，蔡美.蔡美治疗膀胱癌经验［J］.湖南中医杂志，2019，35（4）：32-33.

胡小丽.膀胱癌术后应用消癥肾气汤结合中医埋针治疗效果评价［J］.四川中医，2022，40（9）：141-144.

方雨潇，王淑美，刘伶俐，等.基于HIF-1α/VEGFA信号通路探讨柴胡桂枝汤对三阴性乳腺癌细胞的影响［J］.中国实验方剂学杂志，2023，29（4）：18-24.

朱燃培，桑天庆，谢辉，等.大柴胡汤在消化系统肿瘤中的研究进展［J］.中国实验方剂学杂志，2024，doi：10.13422/j.cnki.syfjx.20232321http://dx.doi.org/10.13422/j.cnki.syfjx.20232321.

程海霞，段清珍.柴胡皂苷D抗肿瘤作用机制研究进展［J］.山西中医，2021，37（2）：57-60.

常彦祥，孙利平.柴胡皂苷D对肝癌大鼠肿瘤生长和免疫功能的影响［J］.中国临床药理学杂志，2020，36（14）：2059-2062.

吴红伟，王临艳，李东辉，等.柴胡皂苷类化合物的药理作用研究进展［J］.中兽医医药杂志，2020，39（6）：35-39.

ZHOU P，SHI W，HE X Y，et al.Saikosaponin D： Review on the antitumour effects，toxicity and pharmacokinetics［J］.Pharm Biol，2021，59（1）：1480-1489.

ZHONG D，ZHANG H J，JIANG Y D，et al.Saikosaponin-d：A potential chemotherapeutics in castration resistant prostate cancer by suppressing cancer metastases and cancer stem cell phenotypes［J］.Biochem Biophys Res Commun，2016，474（4）：722-729.

HU J，LI P，SHI B，et al.Effects and mechanisms of saikosaponin D improving the sensitivity of human gastric cancer cells to cisplatin［J］.ACS Omega，2021，6（29）：18745-18755.

张静，于慧玲，赵鹏伟，等.柴胡皂苷d通过Akt/mTOR信号通路调控胶质瘤C6细胞自噬［J］.中国癌症防治杂志，2022，14（6）：606-610.

冯杰鑫，吴雄，陈建辉，等.柴胡皂苷D通过下调JAK2/STAT3通路抑制乳腺癌细胞的增殖并促进其凋亡［J］.第三军医大学学报，2021，43（9）：852-857.

CHEN L L，XIA L Y，ZHANG J P，et al.Saikosaponin D alleviates cancer cachexia by directly inhibiting STAT3［J］.Phytother Res，2023，37（3）：809-819.

刘宏伟，朱奕，熊洪，等.柴胡皂苷D通过调控上皮间质转化抑制膀胱癌细胞迁移与侵袭的研究［J］.广东医科大学学报，2022，40（5）：486-491.

谢成志，任建勋.中药及复方功效的网络药理学研究与思考［J］.中国实验方剂学杂志，2024，30（1）：198-207.

严易之，李超亚，邓曼菲，等.基于关键病理生理学过程的网络药理学分析新思路［J］.中国实验方剂学杂志，2023，29（17）：203-211.

FATHI N，RASHIDI G，KHODADADI A，et al.STAT3 and apoptosis challenges in cancer［J］.Int J Biol Macromol，2018，117：993-1001.

YOU L，WANG Z，LI H，et al.The role of STAT3 in autophagy［J］.Autophagy，2015，11（5）：729-739.

EL-TANANI M，AL KHATIB A O ，ALADWAN S M，et al.Importance of STAT3 signalling in cancer， metastasis and therapeutic interventions［J］.Cell Signal，2022，92：110275.

YU H，PARDOLL D，JOVE R.STATs in cancer inflammation and immunity：A leading role for STAT3［J］.Nat Rev Cancer，2009，9（11）：798-809.

FUKUOKAYA W，KIMURA T，MIKI J，et al.Effectiveness of intravesical doxorubicin immediately following resection of primary non-muscle-invasive bladder cancer： A propensity score-matched analysis［J］.Clin Genitourin Cancer，2020，18（2）：e55-e61.

JO M J，JEONG S，YUN H K，et al.Genipin induces mitochondrial dysfunction and apoptosis via downregulation of STAT3/mcl-1 pathway in gastric cancer［J］.BMC Cancer，2019，19（1）：739.

刘嘉，严宝飞，许晨新，等.积雪草酸调控JAK2/STAT3信号通路对前列腺癌细胞凋亡的影响［J］.药物评价研究，2023，46（1）：85-91.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect and Mechanism of Zuojinwan on DSS-induced Ulcerative Colitis

Mechanism of Didangtang in Treatment of Bladder Cancer Based on Network Pharmacology

Bioinformatics Reveals Mechanism of Zuoguiwan in Treating Polycystic Ovary Syndrome

Treatment of Chronic Urticaria with Traditional Chinese Medicine by Regulating PI3K/Akt Molecular Pathway： A Review

Related Author

ZUO Ling

ZHANG Zhao

YANG Ju

WANG Jiawei

XIE Shuying

WU Wenbin

JING Manyi

LANG Liwei

Related Institution

The Second Affiliated Hospital of Guangdong Medical University

Chengdu University of Traditional Chinese Medicine

The Fifth Medical Center of Chinese PLA General Hospital

Southern Medical University

Central Military Commission General Administration Bureau of Military Affairs

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰