Effect of total flavonoids from<italic style="font-style: italic"> Clerodendranthus Spicatus</italic> on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis

LIAO Hongliang; GUO Yuntao; TANG Shuainan; CHEN Shuangjian; ZHU Yun; ZHAO Dezhi; JIANG Xin; MA Duan

doi:10.16306/j.1008-861x.2023.03.009

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Effect of total flavonoids from Clerodendranthus Spicatus on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis

Experiment Research | 更新时间：2023-06-03

- Effect of total flavonoids from Clerodendranthus Spicatus on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis
- Academic Journal of Shanghai University of Traditional Chinese Medicine Vol. 37, Issue 3, Pages: 48-56(2023)
- 作者机构：
  
  1.江西中医药大学（江西　南昌　330103）
  2.中科基因生物科技（江苏）有限公司（江苏　南通　226133）
  3.长三角药物高等研究院（江苏　南通　226133）
- 作者简介：
- 基金信息：
- DOI：10.16306/j.1008-861x.2023.03.009
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廖宏亮,郭云涛,唐帅男等.猫须草总黄酮对人肾透明细胞癌786⁃O细胞凋亡的影响及转录组学分析[J].上海中医药大学学报,2023,37(03):48-56.

LIAO Hongliang,GUO Yuntao,TANG Shuainan,et al.Effect of total flavonoids from Clerodendranthus Spicatus on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2023,37(03):48-56.
廖宏亮,郭云涛,唐帅男等.猫须草总黄酮对人肾透明细胞癌786⁃O细胞凋亡的影响及转录组学分析[J].上海中医药大学学报,2023,37(03):48-56. DOI： 10.16306/j.1008-861x.2023.03.009.

LIAO Hongliang,GUO Yuntao,TANG Shuainan,et al.Effect of total flavonoids from Clerodendranthus Spicatus on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2023,37(03):48-56. DOI： 10.16306/j.1008-861x.2023.03.009.

摘要

目的,2,探讨猫须草总黄酮对人肾透明细胞癌786-O细胞凋亡的影响及其机制。,方法,2,用不同浓度（0、25、50、100、200、400 µg/mL）的猫须草总黄酮干预人肾透明细胞癌786-O细胞，采用CCK-8法检测细胞活力，采用膜联蛋白V/碘化丙啶（Annexin V/PI）染色检测细胞凋亡。786-O细胞分别用0、139.9 µg/mL猫须草总黄酮干预后，采用二代高通量测序平台（Illumina Novaseq 6000）对细胞进行RNA-seq建库测序分析，获得通路相关基因，并采用RT-qPCR和Western blot进行验证。,结果,2,①CCK-8检测结果表明，猫须草总黄酮处理人肾透明细胞癌786-O细胞后，细胞活力呈时间及浓度依赖性降低；猫须草总黄酮呈浓度依赖性地促进786-O细胞凋亡。②通过转录组测序共鉴定出2 047个差异基因，其中有773个上调，1 274个下调。基因本体论（GO）富集分析显示，差异基因主要涉及细胞应激反应、凋亡信号通路调控、外源性凋亡信号通路等过程；京都基因和基因组百科全书（KEGG）富集分析显示，差异基因主要涉及细胞凋亡、代谢等。③通过RT-qPCR和Western blot验证几个相关基因的mRNA和蛋白表达水平，与转录组测序的趋势一致（,P,<,0.01）。,结论,2,通过RNA-seq测序数据分析，发现猫须草总黄酮可能通过肿瘤抑制蛋白p53（p53）、丝裂原活化蛋白激酶（MAPK）、核因子-κB（NF-κB）等信号通路诱导人肾透明细胞癌786-O细胞凋亡。

Abstract

Objective： To explore the mechanism of total flavonoids from, Clerodendranthus Spicatus, on the apoptosis of human renal clear cell carcinoma 786-O cells.,Methods,2,Human renal clear cell carcinoma 786-O cells were treated with different concentrations （0， 25， 50， 100， 200， 400 µg/mL） of total flavonoids from ,Clerodendranthus Spicatus,. Cell viability was detected by CCK-8 assay， and cell apoptosis was measured by Annexin V/propidium iodide （PI） staining. RNA-seq library sequencing analysis was carried out through Illumina Novaseq 6000 platform on cells treated with 0 and 139.9 µg/mL of total flavonoids from ,Clerodendranthus Spicatu,. Pathway-related genes were obtained and verified by RT-qPCR and Western blot.,Results,2,①CCK-8 results showed that the viability of human renal clear cell carcinoma 786-O cells decreased in a time and concentration-dependent manner after treated with total flavonoids from ,Clerodendranthus Spicatu,. And,total flavonoids from ,Clerodendranthus Spicatu, induced apoptosis of 786-O cells in a concentration-dependent manner. ②A total of 2 047 differential genes were identified by transcriptome sequencing， among which 773 were up-regulated and 1 274 were down-regulated. Gene Ontology （GO） enrichment analysis showed that the differential genes were mainly involved in processes such as cellular stress response， apoptotic signaling pathway regulation， and exogenous apoptotic signaling pathway. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis showed that the differential genes were mainly involved in cell apoptosis and metabolism. ③The mRNA and protein expression levels of several related genes were verified by RT-qPCR and Western blot， which was consistent with the trend of transcriptome sequencing （,P,<,0.01）.,Conclusion,2,RNA-seq sequencing data analysis showed that total flavonoids from ,Clerodendranthus Spicatu, may induce apoptosis of human renal clear cell carcinoma 786-O cells through signaling pathways such as tumor suppressor protein p53 （p53）， mitogen-activated protein kinase （MAPK）， and nuclear factor-κB （NF-κB）.

关键词

肾茶猫须草总黄酮肾透明细胞癌细胞凋亡转录组测序信号通路

Keywords

Clerodendranthus Spicatutotal flavonoids from Clerodendranthus Spicaturenal clear cell carcinomaapoptosistranscriptome sequencingsignaling pathway

references

LIOU Y，CHEN P，CHU S，et al. Thymoquinone suppresses the proliferation of renal cell carcinoma cells via reactive oxygen species-induced apoptosis and reduces cell stemness ［J］. Environ Toxicol， 2019， 34（11）： 1208-1220.

福建省中医药研究院. 福建药物志［M］. 福州：福建科学技术出版社，1994： 390.

Fujian Academy of Traditional Chinese Medicine. Records of Fujian medicines ［M］. Fuzhou： Fujian Science and Technology Press， 1994： 390.

姜帅，邹德志，肖培根，等. 肾茶的传统应用调查与研究进展［J］. 中国现代中药，2015， 17（9）： 980-987.

JIANG S， ZOU D Z，XIAO P G， et al. Investigation and research progress of Traditional application of kidney tea ［J］. Chinese Modern Traditional Chinese Medicine， 2015， 17（9）： 980-987.

许娜，许旭东，杨峻山. 猫须草的研究进展［J］. 中草药，2010， 41（5）： 848-852.

XU N， XU X D， YANG J S. Research Progress of Cat’s Whiskers ［J］. Chinese Herbal Medicine， 2010， 41（5）： 848-852.

张荣荣. 海南栽培肾茶化学成分及其生物活性研究［D］. 海口：海南大学，2017.

ZHANG R R. Study on chemical constituents and biological activities of cultivated kidney tea in Hainan ［D］. Haikou： Hainan University， 2017.

孙照翠，郑庆霞，吴海峰，等. 猫须草水溶性化学成分的研究［J］. 中国药学杂志，2014， 49（1）： 22-25.

SUN Z C，ZHENG Q X， WU H F， et al. Study on the chemical constituents of water soluble in Catatophora chinensis ［J］. Chinese Journal of Pharmacy， 2014， 49（1）： 22-25.

张永怡，吴家超，林青华，等. 民族药肾茶的化学成分与药理作用研究进展［J］. 中医药学报，2021， 49（1）： 112-120.

ZHANG Y Y， WU J C， LIN Q H，et al. Research progress on the chemical constituents and pharmacological effects of Shencha ［J］. Chinese Journal of Traditional Chinese Medicine， 2021， 49（1）：112-120.

LONG H M， LUO Y， CHENG H Y， et al. Anti-renal cancer activity of total flavonoids from kidney tea ［J］. Journal of Gannan Medical College， 2017， 37（2）： 179-184.

TEZUKA Y，STAMPOULIS P，BANSKOTA A H，et al. Constituents of the Vietnamese medicinal Plant Orthosiphon stamineus［J］. Chem Pharm Bull， 2000， 48： 1711-1719.

赵爱华，赵勤实，李蓉涛，等. 肾茶的化学成分［J］. 云南植物研究，2004， 26： 563-568.

ZHAO A H，ZHAO Q S，LI R T，et al. Chemical constituents of kidney tea ［J］. Plant Research of Yunnan， 2004， 26： 563-568.

马知伊. 肾茶叶挥发油化学成分分析［J］. 韩山师范学院学报， 2013， 34（3）： 50-54，59.

MA Z Y. Analysis on Chemical Constituents of Volatile oil of Kidney Tea ［J］. Journal of Hanshan Normal University， 2013， 34（3）： 50-54，59.

谭俊杰，谭昌恒，陈伊蕾，等. 肾茶化学成分的研究［J］. 天然产物研究与开发，2009， 21（4）： 608-611， 592.

TAN J J，TAN C H，CHEN Y L，et al. Study on Chemical Constituents of Kidney Tea ［J］. Natural Products Research and Development， 2009， 21（4）： 608-611，592.

AWALE S，TEZUKA Y，BANSKOTA A H，et a1. Highly-oxy-genated isopimarane-type diterpenes from Orthosiphon stamineus of indonesia and their nitric oxide inhibitory activity［J］. Chem Pharm Bull， 2003， 51： 268-275.

OLAH N K，RADU L， MOGOSAN C， et a1. Phytochemical and pharmacological studies on Orthosiphon stamineus Benth. hydroalcoholic extracts［J］. J Pharm Biomed Anal，2003，33：117-123.

AWALE S， TEZUKA Y， BAMKOTA A H，et a1. Nitric oxide inhibi-tory isopiarmmme-type diterpenes from Orthosiphon stanfineus of Indonesia［J］. J Nat Prod， 2003， 66： 255-258.

NGUYEN M T R，AWALE S，TEZUKA Y，et a1. Staminane-and isopimatane-type diterpenes from Orthosiphon stamineus of Taiwan and their nitric oxide inhibitory activity［J］. J Nat Pmd. 2004， 67（4）： 654-658.

张荣荣，梅文莉，黄圣卓，等. 海南栽培肾茶的化学成分研究［J］. 热带亚热带植物学报，2017， 25（2）： 182-188.

ZHANG R R，MEI W L，HUANG S Z，et al. Study on the chemical constituents of Cultivated kidney Tea in Hainan ［J］. Journal of Tropical and Subtropical Botany，2017，25（2）：182-188.

王敏，梁敬钰，陈雪英. 肾茶的水溶性成分［J］. 中国天然药物，2007， 5（1）： 27-30.

WANG M， LIANG J Y， CHEN X Y. Water soluble components of kidney tea ［J］. Chinese Natural Medicine， 2007， 5（1）： 27-30.

王锐，李云然. 肾茶的化学成分、生物活性及临床应用研究进展［J］. 广东化工，2020， 47（3）： 92-93，100.

WANG R， LI Y R. Advances in Studies on Chemical Constituents， Bioactivities， and Clinical application of Clerodendranthus Spicatus ［J］. Guangdong Chemical Industry， 2020， 47（3）： 92-93， 100.

MANTHEY J A，GUTHRIE N，GROHMANN K. Biological properties of citrus flavonoids pertaining to Cancer and inflammation［J］. Curt Med Chem， 2001， 8（2）： 135-153.

MIDDLETON E，KANDASWAMI C，THEOHARIDES T C. The effects of plant flavonoids on mammalian cells： implications for inflammation heart disease and cancer［J］. Pharmacol Rev， 2000， 52（4）： 673-751.

GARCIA-LAFUENTE A，GUILLAMN E，VILLARES A，et a1. Flavonoids as anti-inflammatory agents： implications in cancer and cardiovascular disease［J］. Inflamm Res， 2009， 58（9）： 537-552.

曾佑炜. 黄酮抗癌作用研究进展［J］. 天然产物研究与开发，2016， 28（11）： 1838-1844.

ZENG Y W. Research Progress of anticancer effect of flavonoids ［J］. Natural Products Research and Development， 2016， 28（11）： 1838-1844.

朱晓芹，郑雅，刘志强. 半枝莲主要黄酮类成分抗癌协同机制的网络药理学分析［J］. 中药新药与临床药理，2020， 31（9）： 1037-1044.

ZHU X Q， ZHENG Y， LIU Z Q. Network pharmacological analysis of anticancer synergistic mechanism of main flavonoids in Schistoderma slichoides［J］. New Chinese Medicine and Clinical Pharmacology， 2020， 31（9）： 1037-1044.

ZAIN W N，RAHMAT A， OTHMAN F， et a1. Antiproliferative properties of Clausine-B against cancer cell lines［J］. Malays J Med Sci， 2009， 16（3）： 31-36.

LAI C S， TSAI M L， CHENG A C， et a1. Chemoprevention of colonic tumorigenesis by dietary hydroxylated polyme-thoxyflavones in azoxymethane-treated mice［J］. Mol Nutr Food Res， 2010， 55（2）： 278-290.

HOSSAIN M A，ISMAIL Z，RAHMAN A， et a1. Chemical corn. position and antifungal properties of the essential oils and crude extracts of Orthosiphon stamineus Benth［J］. Ind Crop Prod，2008， 27： 328-334.

谢丽华，樊星，李泽君，等. 转录组学技术在中药肝毒性研究中的应用［J］. 中草药，2015， 46（10）： 1536-1541.

XIE L H， FAN X， LI Z J， et al.Application of Transcriptomics in the Study of hepatotoxicity of Traditional Chinese Medicine ［J］. Chinese Herbal Medicine， 2015， 46（10）： 1536-1541.

WEI W， WANG Y， ZHANG Q， et al. Global characterization of Artemisia annua glandular triehome trmmeriptome using 454 pyrosequeneing［J］. BMC Genomics， 2009， 10： 465.

ZENG S， XIAN G， GUO J，et a1. Development of a EST dataset and characterization of EST-SSRs in a traditional Chinese medicinal plant， Epimedium sagittatum （Sieb. Et Zucc.） Maxim ［J］. BMC Genomics， 2010， 11： 94.

吴宏清，王磊，陶美华，等. 化学诱导后白木香转录组文库的构建与测序［J］. 生物技术通报，2013，（8）： 63-67.

WU H Q，WANG L，TAO M H，et al. Transcriptome Library Construction and Sequencing from Chemically Induced Aquilaria sinensis ［J］. Biotechnology Bulletin， 2013，（8）： 63-67.

BOS C L，KODACH L L，VANDEN BRINK G R， et a1. Effect of aspirin on the Wnt/B-catenin pathway is mediated via protein phosphatase 2A［J］. Oncogene， 2006， 25（49）： 6447-6456.

CAMPBELL L，NUTTALL R，GRIFFITHS D，et a1. Activated extracellular signal-regulated kinase is an independent prognostic factor in clinically confined renal cell carcinoma. Cancer， 2009： 115（15）： 3457-3467.

HUANG D，DING Y，LUO W M. Inhibition of MAPK kinase signaling pathways suppressed renal cell carcinoma growth and angiogenesis in vivo［J］. Cancer Res， 2008： 68： 81-88.

DENG J. How to unleash mitochondrial apoptotic block ades to kill cancers？［J］. Acta Pharm Sin B， 2017， 7（1）： 18-26.

YING Y，PADANILAM B J. Regulation of necrotic cell death： p53 PARPl and cyclophilin Doverlapping pathways of regulated necrosis？［J］. Cell Mol Life Sci， 2016， 73（11-12）： 2309-2324.

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Effect of total flavonoids from Clerodendranthus Spicatus on apoptosis of human renal clear cell carcinoma 786⁃O cells and transcriptomic analysis

DOI：10.16306/j.1008-861x.2023.03.009

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