FOLLOWUS
- Share with wechat friends or circle of friends
吴茱萸碱联合小檗碱对CCD-18Co诱导的结肠上皮细胞表型转化的调节效应
- Affiliations：
1. 1Department of Traditional Chinese Medicine, Affiliated Bao’an Hospital of Shenzhen, Southern Medical University, Shenzhen 518100, China
2. 2Community Healthy Service Center, Shenzhen Bao’an Traditional Chinese Medicine Hospital Group, Shenzhen 518100, China
- Author bio：
* Tel: 86-755-27788311, E-mail: huangchao06@163.com
- Funds：National Natural Science Foundation of China(81903985, the running period is 2020.1-2022.12);Natural Science Foundation of Guangdong Province(2018A030310060, the running period is 2018.6-2021.5);Postdoctoral Research Foundation of China(2018M643353, the running period is 2018.9-2019.7)
DOI：10.24920/003670
中图分类号：收稿日期：2019-11-22，
网络出版日期：2020-09-30，
纸质出版日期：2020-09-30
- Accepted：
Scan QR Code
黄超, 向科明, 梁炳君, 等. 吴茱萸碱联合小檗碱对CCD-18Co诱导的结肠上皮细胞表型转化的调节效应[J]. 中国医学科学杂志（英文版）, 2020,35(3):195-206.
Huang Chao, Xiang Keming, Liang Bingjun, et al. Combination of Evodiamine with Berberine Reveals a Regulatory Effect on the Phenotypic Transition of Colon Epithelial Cells Induced by CCD-18Co[J]. Chinese medical sciences journal, 2020, 35(3): 195-206.
黄超, 向科明, 梁炳君, 等. 吴茱萸碱联合小檗碱对CCD-18Co诱导的结肠上皮细胞表型转化的调节效应[J]. 中国医学科学杂志（英文版）, 2020,35(3):195-206. DOI： 10.24920/003670.
Huang Chao, Xiang Keming, Liang Bingjun, et al. Combination of Evodiamine with Berberine Reveals a Regulatory Effect on the Phenotypic Transition of Colon Epithelial Cells Induced by CCD-18Co[J]. Chinese medical sciences journal, 2020, 35(3): 195-206. DOI： 10.24920/003670.
摘要
目的
基质细胞之间或基质细胞与癌细胞之间存在着细胞转分化过程。吴茱萸碱和小檗碱是中药方剂左金丸的主要药理成分
二者在肿瘤微环境重塑中起着至关重要的作用。本研究旨在探讨吴茱萸碱联合小檗碱（cBerEvo）对肿瘤相关成纤维细胞诱导的结肠上皮细胞表型转化的影响及其相关机制。
方法
采用制备好的人结肠肌成纤维细胞系CCD-18Co条件化培养基诱导正常人结肠上皮细胞系HCoEpiCs向间质细胞转化
同时观察TGF-β受体抑制剂LY364947或cBerEvo对HCoEpiCs细胞转化是否有抑制作用。用相差显微镜观察HCoEpiCs形态学变化。免疫荧光显微镜观察及检测上皮细胞-间充质转化标记物包括E-钙粘蛋白（E-cadherin）
波形蛋白（vimentin）以及α-平滑肌肌动蛋白（α-SMA）的表达。创伤愈合试验评估细胞迁移情况。Western blotting检测E-cadherin、vimentin、α-SMA、Snail、ZEB1及Smads蛋白表达。
结果
与对照组细胞相比
经肿瘤相关成纤维细胞样CCD-18Co细胞诱导的HCoEpiCs细胞表达E-cadherin下调、表达vimentin、α-SMA、Snail、ZEB1上调（
P
<
0.05）
同时HCoEpiCs细胞迁移能力提高（
P
<
0.05）
表达Smads蛋白包括Smad2、Smad3、p-Smad2、p-Smad3和Smad4上调。TGF-B抑制剂LY364947或吴茱萸碱联合小檗碱能够以浓度依赖的方式部分逆转上述上述CCD-18Co细胞的诱导作用。此外
cBerEvo能以浓度依赖性的方式抑制p-Smad2/Smad2及p-Smad3/Smad3的比例。
结论
本研究结果表明cBerEvo通过抑制TGF-β/Smads信号通路的激活调节由CCD-18Co细胞诱导的结肠上皮细胞转分化过程。
Abstract
Objective
Transdifferentiation exists between stromal cells or between stromal cells and cancer cells. Evodiamine and berberine are predominant pharmacological components of
Zuojin
pill
a prescription of Traditional Chinese Medicine
playing crucial functions in remolding of tumor microenvironment. This study aimed to explore the effect of combination of evodiamine with berberine (cBerEvo) on the phenotypic transition of colon epithelial cells induced by tumor-associated fibroblasts
as well as the involved mechanisms.
Methods
Human normal colon epithelial cell line HCoEpiC cells were treated with the prepared conditioned medium of CCD-18Co
a human colon myofibroblast line
to induce epithelial-mesenchymal transition. Phase contrast microscope was used to observe the morphological changes. Epithelial-mesenchymal transition markers including E-cadherin
vimentin and alpha-smooth muscle actin (α-SMA) were observed with immunofluorescence microscopy. Migration was assessed by wound healing assay. Western blotting was used to detect the expressions of E-cadherin
vimentin
α-SMA
Snail
ZEB1 and Smads.
Results
In contrast to the control
the tumor-associated fibroblasts-like CCD-18Co cells induced down-regulation of E-cadherin and up-regulation of vimentin
α-SMA
Snail and ZEB1 (
P
<
0.05)
and promoted migration of HCoEpiCs (
P
<
0.05)
with over expression of Smads including Smad2
p-Smad2
Smad3
p-Smad3 and Smad4 (
P
<
0.05)
which were abolished by a transforming growth factor-β (TGF-β) receptor inhibitor LY364947 and by cBerEvo in a concentration dependent manner. In addition
cBerEvo-inhibited ratios of p-Smad2/Smad2 and p-Smad3/Smad3 were also dose dependent.
Conclusion
The above results suggest that cBerEvo can regulate the differentiation of colon epithelial cells induced by CCD-18Co through suppressing activity of TGF-β/Smads signaling pathway.
关键词
Keywords
references
Eble JA , Niland S . The extracellular matrix in tumor progression and metastasis . Clin Exp Metastasis 2019 ; 36 ( 3 ): 171 - 98 . doi: 10.1007/s10585-019-09966-1 10.1007/s10585-019-09966-1 . DOI: 10.1007/s10585-019-09966-1 http://doi.org/10.1007/s10585-019-09966-1 https://www.ncbi.nlm.nih.gov/pubmed/30972526 https://www.ncbi.nlm.nih.gov/pubmed/30972526
Metzler VM , Pritz C , Riml A , et al . Separation of cell survival, growth, migration, and mesenchymal transdifferentiation effects of fibroblast secretome on tumor cells of head and neck squamous cell carcinoma . Tumour Biol 2017 ; 39 ( 11 ): 1010428317705507 . doi: 10.1177/1010428317705507 10.1177/1010428317705507 . https://www.ncbi.nlm.nih.gov/pubmed/29157176 https://www.ncbi.nlm.nih.gov/pubmed/29157176
Zhang Y , Cai H , Chen S , et al . Exosomal transfer of miR-124 inhibits normal fibroblasts to cancer-associated fibroblasts transition by targeting sphingosine kinase 1 in ovarian cancer . J Cell Biochem 2019 ; 120 ( 8 ): 13187 - 201 . doi: 10.1002/jcb.28593 10.1002/jcb.28593 . https://www.ncbi.nlm.nih.gov/pubmed/30957275 https://www.ncbi.nlm.nih.gov/pubmed/30957275
Whatcott C , Han H , Posner RG , et al . Tumor-stromal interactions in pancreatic cancer . Crit Rev Oncog 2013 ; 18 ( 1-2 ): 135 - 51 . doi: 10.1615/critrevoncog.v18.i1-2.80 10.1615/critrevoncog.v18.i1-2.80 . DOI: 10.1615/critrevoncog.v18.i1-2.80 http://doi.org/10.1615/critrevoncog.v18.i1-2.80 https://www.ncbi.nlm.nih.gov/pubmed/23237556 https://www.ncbi.nlm.nih.gov/pubmed/23237556
Kahata K , Dadras MS , Moustakas A . TGF-β family signaling in epithelial differentiation and epithelial-mesenchymal transition . Cold Spring Harb Perspect Biol 2018 ; 10 ( 1 ). pii: a022194 . doi: 10.1101/cshperspect.a022194 10.1101/cshperspect.a022194 . DOI: 10.1101/cshperspect.a022194 http://doi.org/10.1101/cshperspect.a022194 https://www.ncbi.nlm.nih.gov/pubmed/28246184 https://www.ncbi.nlm.nih.gov/pubmed/28246184
Liu Y , Xue M , Du S , et al . Competitive endogenous RNA is an intrinsic component of EMT regulatory circuits and modulates EMT . Nat Commun 2019 ; 10 ( 1 ): 1637 . doi: 10.1038/s41467-019-09649-1 10.1038/s41467-019-09649-1 . https://www.ncbi.nlm.nih.gov/pubmed/30967542 https://www.ncbi.nlm.nih.gov/pubmed/30967542
Wang QS , Zhu XN , Jiang HL , et al . Protective effects of alginate-chitosan microspheres loaded with alkaloids from Coptis chinensis Franch. and Evodia rutaecarpa (Juss.) Benth. (Zuojin Pill) against ethanol-induced acute gastric mucosal injury in rats . Drug Des Devel Ther 2015 ; 9 : 6151 - 65 . doi: 10.2147/DDDT.S96056 10.2147/DDDT.S96056 . DOI: 10.2147/DDDT.S96056 http://doi.org/10.2147/DDDT.S96056 https://www.ncbi.nlm.nih.gov/pubmed/26640368 https://www.ncbi.nlm.nih.gov/pubmed/26640368
Huang C , Tao L , Wang XL , et al . Berberine reversed the epithelial-mesenchymal transition of normal colonic epithelial cells induced by SW480 cells through regulating the important components in the TGF-β pathway . J Cell Physiol 2019 ; 234 ( 7 ): 11679 - 91 . doi: 10.1002/jcp.27835 10.1002/jcp.27835 . https://www.ncbi.nlm.nih.gov/pubmed/30536375 https://www.ncbi.nlm.nih.gov/pubmed/30536375
Yang W , Gong X , Wang X , et al . A mediator of phosphorylated Smad2/3, evodiamine, in the reversion of TAF-induced EMT in normal colonic epithelial cells . Invest New Drugs 2019 ; 37 ( 5 ): 865 - 75 . doi: 10.1007/s10637-018-0702-x 10.1007/s10637-018-0702-x . https://www.ncbi.nlm.nih.gov/pubmed/30488243 https://www.ncbi.nlm.nih.gov/pubmed/30488243
Hong D , Liu T , Huang W , et al . Gremlin1 delivered by mesenchymal stromal cells promoted epithelial-mesenchymal transition in human esophageal squamous cell carcinoma . Cell Physiol Biochem 2018 ; 47 ( 5 ): 1785 - 99 . doi: 10.1159/000491060 10.1159/000491060 . DOI: 10.1159/000491060 http://doi.org/10.1159/000491060 https://www.ncbi.nlm.nih.gov/pubmed/29953975 https://www.ncbi.nlm.nih.gov/pubmed/29953975
Aiello NM , Kang Y . Context-dependent EMT programs in cancer metastasis . J Exp Med 2019 ; 216 ( 5 ): 1016 - 26 . doi: 10.1084/jem.20181827 10.1084/jem.20181827 . DOI: 10.1084/jem.20181827 http://doi.org/10.1084/jem.20181827 https://www.ncbi.nlm.nih.gov/pubmed/30975895 https://www.ncbi.nlm.nih.gov/pubmed/30975895
Feng Y , Li NX , Yin HL , et al . Thermo- and pH-responsive, lipid-coated, mesoporous silica nanoparticle-based dual drug delivery system to improve the antitumor effect of hydrophobic drugs . Mol Pharm 2019 ; 16 ( 1 ): 422 - 36 . doi: 10.1021/acs.molpharmaceut.8b01073 10.1021/acs.molpharmaceut.8b01073 . DOI: 10.1021/acs.molpharmaceut.8b01073 http://doi.org/10.1021/acs.molpharmaceut.8b01073 https://www.ncbi.nlm.nih.gov/pubmed/30525641 https://www.ncbi.nlm.nih.gov/pubmed/30525641
Xu J , Song Z , Guo Q , et al . Synergistic effect and molecular mechanisms of Traditional Chinese Medicine on regulating tumor microenvironment and cancer cells . Biomed Res Int 2016 ; 2016 : 1490738 . doi: 10.1155/2016/1490738 10.1155/2016/1490738 . https://www.ncbi.nlm.nih.gov/pubmed/27042656 https://www.ncbi.nlm.nih.gov/pubmed/27042656
Lu P , Su W , Miao ZH , et al . Effect and mechanism of ginsenoside Rg3 on postoperative life span of patients with non-small cell lung cancer . Chin J Integr Med 2008 ; 14 ( 1 ): 33 - 6 . doi: 10.1007/s11655-007-9002 10.1007/s11655-007-9002 . DOI: 10.1007/s11655-007-9002-6 http://doi.org/10.1007/s11655-007-9002-6 https://www.ncbi.nlm.nih.gov/pubmed/18219455 https://www.ncbi.nlm.nih.gov/pubmed/18219455
Wang B , Cui J . Treatment of mid-late stage NSCLC using sodium cantharidinate/vitamin B6/GP regimen in clinic . J Cancer Res Ther 2014 ; 10 ( Suppl 1 ): C79 - 81 . doi: 10.4103/0973-1482.139771 10.4103/0973-1482.139771 .
James MI , Iwuji C , Irving G , et al . Curcumin inhibits cancer stem cell phenotypes in ex vivo models of colorectal liver metastases, and is clinically safe and tolerable in combination with FOLFOX chemotherapy . Cancer Lett 2015 ; 364 ( 2 ): 135 - 41 . doi: 10.1016/j.canlet.2015.05.005 10.1016/j.canlet.2015.05.005 . DOI: 10.1016/j.canlet.2015.05.005 http://doi.org/10.1016/j.canlet.2015.05.005 https://www.ncbi.nlm.nih.gov/pubmed/25979230 https://www.ncbi.nlm.nih.gov/pubmed/25979230
Liu Y , Jia Z , Dong L , et al . A randomized pilot study of atractylenolide I on gastric cancer cachexia patients . Evid Based Complement Alternat Med 2008 ; 5 ( 3 ): 337 - 44 . doi: 10.1093/ecam/nem031 10.1093/ecam/nem031 . DOI: 10.1093/ecam/nem031 http://doi.org/10.1093/ecam/nem031 https://www.ncbi.nlm.nih.gov/pubmed/18830451 https://www.ncbi.nlm.nih.gov/pubmed/18830451
Xiang Y , Guo Z , Zhu P , et al . Traditional Chinese medicine as a cancer treatment: modern perspectives of ancient but advanced science . Cancer Med 2019 ; 8 ( 5 ): 1958 - 75 . doi: 10.1002/cam4.2108 10.1002/cam4.2108 . DOI: 10.1002/cam4.2108 http://doi.org/10.1002/cam4.2108 https://www.ncbi.nlm.nih.gov/pubmed/30945475 https://www.ncbi.nlm.nih.gov/pubmed/30945475
Li W , Wang Q , Su Q , et al . Honokiol suppresses renal cancer cells’ metastasis via dual-blocking epithelial-mesenchymal transition and cancer stem cell properties through modulating miR-141/ZEB2 signaling . Mol Cells 2014 ; 37 ( 5 ): 383 - 8 . doi: 10.14348/molcells.2014.0009 10.14348/molcells.2014.0009 . DOI: 10.14348/molcells.2014.0009 http://doi.org/10.14348/molcells.2014.0009 https://www.ncbi.nlm.nih.gov/pubmed/24810210 https://www.ncbi.nlm.nih.gov/pubmed/24810210
Chao DC , Lin LJ , Kao ST , et al . Inhibitory effects of Zuo-Jin-Wan and its alkaloidal ingredients on activator protein 1, nuclear factor-κB, and cellular transformation in HepG2 cells . Fitoterapia 2011 ; 82 ( 4 ): 696 - 703 . doi: 10.1016/j.fitote.2011.02.009 10.1016/j.fitote.2011.02.009 . https://www.ncbi.nlm.nih.gov/pubmed/21356280 https://www.ncbi.nlm.nih.gov/pubmed/21356280
Huang C , Wang XL . Zuojin Pill , a classical Traditional Chinese Medicine formula, inhibits the Tafs induced phenotype transition and migration in normal colonic epithelial cells through TGF-β/Smads signaling . Herbal Med 2019 . Available from: http: www.openaccessebooks.com/herbal-medicine.html http: www.openaccessebooks.com/herbal-medicine.html http: www.openaccessebooks.com/herbal-medicine.html
Huang C , Wang XL , Qi FF , et al . Berberine inhibits epithelial-mesenchymal transition and promotes apoptosis of tumour-associated fibroblast-induced colonic epithelial cells through regulation of TGF-β signalling . J Cell Commun Signal 2020 , 14 ( 1 ): 53 - 66 . doi: 10.1007/s12079-019-00525-7 10.1007/s12079-019-00525-7 . DOI: 10.1007/s12079-019-00525-7 http://doi.org/10.1007/s12079-019-00525-7 https://www.ncbi.nlm.nih.gov/pubmed/31399854 https://www.ncbi.nlm.nih.gov/pubmed/31399854
Du J , Sun Y , Lu YY , et al . Berberine and evodiamine act synergistically against human breast cancer MCF-7 cells by inducing cell cycle arrest and apoptosis . Anticancer Res 2017 ; 37 ( 11 ): 6141 - 51 . doi: 10.21873/anticanres.12063 10.21873/anticanres.12063 . DOI: 10.21873/anticanres.12063 http://doi.org/10.21873/anticanres.12063 https://www.ncbi.nlm.nih.gov/pubmed/29061795 https://www.ncbi.nlm.nih.gov/pubmed/29061795
Wang XN , Han X , Xu LN , et al . Enhancement of apoptosis of human hepatocellular carcinoma SMMC-7721 cells through synergy of berberine and evodiamine . Phytomedicine 2008 ; 15 ( 12 ): 1062 - 8 . doi: 10.1016/j.phymed.2008.05.002 10.1016/j.phymed.2008.05.002 . https://www.ncbi.nlm.nih.gov/pubmed/18579357 https://www.ncbi.nlm.nih.gov/pubmed/18579357
Li-Weber M . Targeting apoptosis pathways in cancer by Chinese medicine . Cancer Lett 2013 ; 332 ( 2 ): 304 - 12 . doi: 10.1016/j.canlet.2010.07.015 10.1016/j.canlet.2010.07.015 . DOI: 10.1016/j.canlet.2010.07.015 http://doi.org/10.1016/j.canlet.2010.07.015 https://www.ncbi.nlm.nih.gov/pubmed/20685036 https://www.ncbi.nlm.nih.gov/pubmed/20685036
Liu H , Huang C , Wu L , et al . Effect of evodiamine and berberine on miR-429 as an oncogene in human colorectal cancer . Onco Targets Ther 2016 ; 9 : 4121 - 7 . doi: 10.2147/OTT.S104729 10.2147/OTT.S104729 . DOI: 10.2147/OTT.S104729 http://doi.org/10.2147/OTT.S104729 https://www.ncbi.nlm.nih.gov/pubmed/27462166 https://www.ncbi.nlm.nih.gov/pubmed/27462166
Cho KH , Yu SL , Cho DY , et al . Breast cancer metastasis suppressor 1 (BRMS1) attenuates TGF-β1-induced breast cancer cell aggressiveness through downregulating HIF-1α expression . BMC Cancer 2015 ; 15 : 829 . doi: 10.1186/s12885-015-1864-y 10.1186/s12885-015-1864-y . DOI: 10.1186/s12885-015-1864-y http://doi.org/10.1186/s12885-015-1864-y https://www.ncbi.nlm.nih.gov/pubmed/26520789 https://www.ncbi.nlm.nih.gov/pubmed/26520789
Li Y , Zhu G , Zhai H , et al . Simultaneous stimulation with tumor necrosis factor-α and transforming growth factor-β1 induces epithelial-mesenchymal transition in colon cancer cells via the NF-κB pathway . Oncol Lett 2018 ; 15 ( 5 ): 6873 - 80 . doi: 10.3892/ol.2018.8230 10.3892/ol.2018.8230 . https://www.ncbi.nlm.nih.gov/pubmed/29725419 https://www.ncbi.nlm.nih.gov/pubmed/29725419
Cui Q , Ren J , Zhou Q , et al . Effect of asiatic acid on epithelial-mesenchymal transition of human alveolar epithelium A549 cells induced by TGF-β1 . Oncol Lett 2019 ; 17 ( 5 ): 4285 - 92 . doi: 10.3892/ol.2019.10140 10.3892/ol.2019.10140 . DOI: 10.3892/ol.2019.10140 http://doi.org/10.3892/ol.2019.10140 https://www.ncbi.nlm.nih.gov/pubmed/30988806 https://www.ncbi.nlm.nih.gov/pubmed/30988806
Yu Y , Lin Y , Yang G , et al . The interplay between TGF-β/SMAD and BMP/SMAD signaling pathways in the epithelial mesenchymal transition of A549 cells induced by silica . Toxicol Mech Methods 2018 ; 28 ( 4 ): 286 - 92 . doi: 10.1080/15376516.2017.1407978 10.1080/15376516.2017.1407978 . DOI: 10.1080/15376516.2017.1407978 http://doi.org/10.1080/15376516.2017.1407978 https://www.ncbi.nlm.nih.gov/pubmed/29161937 https://www.ncbi.nlm.nih.gov/pubmed/29161937
Zhang X , He H , Liang D , Protective effects of berberine on renal injury in streptozotocin (STZ)-induced diabetic mice . Int J Mol Sci 2016 ; 17 ( 8 ). pii: E1327 . doi: 10.3390/ijms17081327 10.3390/ijms17081327 . DOI: 10.3390/ijms17081327 http://doi.org/10.3390/ijms17081327 https://www.ncbi.nlm.nih.gov/pubmed/27529235 https://www.ncbi.nlm.nih.gov/pubmed/27529235
Sun SF , Zhao TT , Zhang HJ , et al . Renoprotective effect of berberine on type 2 diabetic nephropathy in rats . Clin Exp Pharmacol Physiol 2015 ; 42 ( 6 ): 662 - 70 . doi: 10.1111/1440-1681.12402 10.1111/1440-1681.12402 . DOI: 10.1111/1440-1681.12402 http://doi.org/10.1111/1440-1681.12402 https://www.ncbi.nlm.nih.gov/pubmed/25867602 https://www.ncbi.nlm.nih.gov/pubmed/25867602
Wei J , Li Z , Yuan F . Evodiamine might inhibit TGF-beta1-induced epithelial-mesenchymal transition in NRK52E cells via Smad and PPAR-gamma pathway . Cell Biol Int 2014 ; 38 ( 7 ): 875 - 80 . doi: 10.1002/cbin.10270 10.1002/cbin.10270 . DOI: 10.1002/cbin.10270 http://doi.org/10.1002/cbin.10270 https://www.ncbi.nlm.nih.gov/pubmed/24604887 https://www.ncbi.nlm.nih.gov/pubmed/24604887
Yang D , Li L , Qian S , et al . Evodiamine ameliorates liver fibrosis in rats via TGF-β1/Smad signaling pathway . J Nat Med 2018 ; 72 ( 1 ): 145 - 54 . doi: 10.1007/s11418-017-1122-5 10.1007/s11418-017-1122-5 . DOI: 10.1007/s11418-017-1122-5 http://doi.org/10.1007/s11418-017-1122-5 https://www.ncbi.nlm.nih.gov/pubmed/28936800 https://www.ncbi.nlm.nih.gov/pubmed/28936800
关联资源
相关文章
相关作者
相关机构
- © 2024 中国医学科学院 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies