Wulong Xiaozheng Wan medicated serum inhibits epithelial-mesenchymal transition in human gastric carcinoma cell line BGC823 by modulation of transforming growth factor-β_1/Smad signaling
|
摘要
OBJECTIVE:To evaluate the effect of Wulong Xiaozheng Wan medicated serum on the epithelial-mesenchymal transition(EMT)of BGC823 cell induced by transforming growth factor-β_1(TGF-β_1)and to explore its mechanism.METHODS:EMT model of BGC823 was stimulated by TGF-β_1.Wulong Xiaozheng Wan medicated serum and LY-364947 were used as intervention.The proliferation and adhesion of BGC823 were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide and flow cytometry was used to detect the apoptosis.The invasion and migration were detected by Transwell.The level of matrix metalloproteins was detected by enzyme-linked immunosorbent assay.The expressions of related proteins and mRNA of EMT marker and TGF-β_1/Smad signal pathway were detected by Western blot and reverse transcription-polymerase chain reaction.RESULTS:Compared with the TGF-β_1 group,Wulong Xiaozheng Wan medicated serum could inhibit the ability of proliferation,heterogeneous adhesion,invasion,and migration.It also promotes apoptosis and homotypic adhesion in BGC823,with a dose-dependent manner.Meanwhile,Wulong Xiaozheng Wan medicated serum could regulate the expression of related proteins and mRNA of TGF-β_1/Smad signaling pathway,and inhibit the expressions of EMT transcription factors and EMT markers.CONCLUSION:Wulong Xiaozheng Wan medicated serum inhibited epithelial-mesenchymal transition by down-regulated the expression of TβRI and the activation of TG F-β_1/Smad signaling pathway.
OBJECTIVE:To evaluate the effect of Wulong Xiaozheng Wan medicated serum on the epithelial-mesenchymal transition(EMT)of BGC823 cell induced by transforming growth factor-β_1(TGF-β_1)and to explore its mechanism.METHODS:EMT model of BGC823 was stimulated by TGF-β_1.Wulong Xiaozheng Wan medicated serum and LY-364947 were used as intervention.The proliferation and adhesion of BGC823 were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide and flow cytometry was used to detect the apoptosis.The invasion and migration were detected by Transwell.The level of matrix metalloproteins was detected by enzyme-linked immunosorbent assay.The expressions of related proteins and mRNA of EMT marker and TGF-β_1/Smad signal pathway were detected by Western blot and reverse transcription-polymerase chain reaction.RESULTS:Compared with the TGF-β_1 group,Wulong Xiaozheng Wan medicated serum could inhibit the ability of proliferation,heterogeneous adhesion,invasion,and migration.It also promotes apoptosis and homotypic adhesion in BGC823,with a dose-dependent manner.Meanwhile,Wulong Xiaozheng Wan medicated serum could regulate the expression of related proteins and mRNA of TGF-β_1/Smad signaling pathway,and inhibit the expressions of EMT transcription factors and EMT markers.CONCLUSION:Wulong Xiaozheng Wan medicated serum inhibited epithelial-mesenchymal transition by down-regulated the expression of TβRI and the activation of TG F-β_1/Smad signaling pathway.
OBJECTIVE:To evaluate the effect of Wulong Xiaozheng Wan medicated serum on the epithelial-mesenchymal transition(EMT)of BGC823 cell induced by transforming growth factor-β_1(TGF-β_1)and to explore its mechanism.METHODS:EMT model of BGC823 was stimulated by TGF-β_1.Wulong Xiaozheng Wan medicated serum and LY-364947 were used as intervention.The proliferation and adhesion of BGC823 were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide and flow cytometry was used to detect the apoptosis.The invasion and migration were detected by Transwell.The level of matrix metalloproteins was detected by enzyme-linked immunosorbent assay.The expressions of related proteins and mRNA of EMT marker and TGF-β_1/Smad signal pathway were detected by Western blot and reverse transcription-polymerase chain reaction.RESULTS:Compared with the TGF-β_1 group,Wulong Xiaozheng Wan medicated serum could inhibit the ability of proliferation,heterogeneous adhesion,invasion,and migration.It also promotes apoptosis and homotypic adhesion in BGC823,with a dose-dependent manner.Meanwhile,Wulong Xiaozheng Wan medicated serum could regulate the expression of related proteins and mRNA of TGF-β_1/Smad signaling pathway,and inhibit the expressions of EMT transcription factors and EMT markers.CONCLUSION:Wulong Xiaozheng Wan medicated serum inhibited epithelial-mesenchymal transition by down-regulated the expression of TβRI and the activation of TG F-β_1/Smad signaling pathway.
引文
1 Siegel RL,Miller KD,Jemal A.Cancer statistics,2016.CA Cancer J Clin 2016;66(1):7-30.
2 Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015.CA Cancer J Clin 2016;66(2):115-132.
3 Huang L,Wu RL,Xu AM.Epithelial-mesenchymal transition in gastric cancer.Am J Transl Res 2015;7(11):2141-2158.
4 Hartgrink HH,Jansen EP,van Grieken NC,et al.Gastric cancer.Lancet 2009;374(9688):477-490.
5 YE X,Weinberg RA.Epithelial-mesenchymal plasticity:a central regulator of cancer progression.Trends Cell Biol2015;25(11):675-686.
6 Heerboth S,Housman G,Leary M,et al.EMT and tumor metastasis.Clin Translat Med 2015;4(1):6
7 Roussos ET,Keckesova Z,Haley JD,et al.AACR special conference on epithelial-mese nchymal transition and cancer progression and treatment.Cancer Res 2010;70(19):7360-7364.
8 Lander R,Nordin K,Labonne C.The F-box protein Ppa is a common regulator of core EMT factors Twist,Snail,Slug,and Sip1.J Cell Biol 2011;194(1):17-25.
9 Franco-Chuaire ML,Magda Carolina SC,Chuaire-Noack L.Epithelial-mesenchymal transition(EMT):principles and clinical impact in cancer therapy.Invest Clin2013;54(2):186-205.
10 Liu X,Li Z,Song Y,et al.AURKA induces EMT by regulating histone modification through Wnt/β-catenin and PI3K/Akt signaling pathway in gastric cancer.Oncotarget2016;7(22):33152-33164.
11 Li W,Li S,Deng L,et al.Decreased MT1-MMP in gastric cancer suppressed cell migration and invasion via regulating MMPs and EMT.Tumour Biol 2015;36(9):6883-6889.
12 Zohn IE,Li Y,Skolnik EY,et al.p38 and a p38-interacting protein are critical for downregulation of E-cadherin during mouse gastrulation.Cell 2006;125(5):957-969.
13 Krafft E,Lybaert P,Roels E,et al.Transforming growth factor beta 1 activation,storage,and signaling pathways in idiopathic pulmonary fibrosis in dogs.J Vet Intern Med2014;28(6):1666-1675.
14 Strand DW,Liang YY,Yang F,et al.TGF-βinduction of FGF-2 expression in stromal cells requires integrated smad 3 and MAPK pathways.Am J Clin Exp Urol 2014;2(3):239-248.
15 Xu J,Lamouille S,Derynck R.TGF-β-induced epithelial to mesenchymal transition.Cell Research 2009;19(2):156-172.
16 Rees JR,Onwuegbusi BA,Save VE,et al.In vivo and in vitro evidence for transforming growth factor-beta1-mediated epithelial to mesenchymal transition in esophageal adenocarcinoma.Cancer Res 2006;66(19):9583-9590.
17 Hasan TN,Lg B,Shafi G,et al.Anti-proliferative effects of organic extracts from root bark of Juglans Regia L.(RB-JR)on MDA-MB-231 human breast cancer cells:role of Bcl-2/Bax,caspases and Tp53.Asian-Pac.J.Cancer Prev2011;12(2):525-530.
18 Liu JX,Di DL,Wei XN,et al.Cytotoxic diarylheptanoids from the pericarps of walnuts(Juglans regia).Planta Med2008;74(7):754-759.
19 Salimi M,Majd A,Sepahdar Z,et al.Cytotoxicity effects of various Juglans regia(walnut)leaf extracts in human cancer cell lines.Pharm Biol 2012;50(11):1416-1422.
20 Huang L,Zhao H,Huang B,et al.Acanthopanax senticosus:review of botany,chemistry and pharmacology.Pharmazie 2011;66(2):83-97.
21 Herszenyi L,Hritz I,Pregun I,et al.Alterations of glutathione S-transferase and matrix metalloproteinase-9 expressions are early events in esophageal carcinogenesis.World JGastroenterology 2007;13(5):676-682.
22 Guo J,Wang B,Fu Z,et al.Hypoxic microenvironment induces EMT and upgrades stem-like properties of gastric cancer cells.Technol Cancer Res Treat 2016;15(1):60-68.
23 Thiery JP,Acloque H,Huang RYJ,et al.Epithelial-mesenchymal transitions in development and disease.Cell 2014;139(5):871-890.
24 Kalluri R,Weinberg RA.The basics of epithelial-mesenchymal transition.J Clin Invest 2009;119(6):1420-1428.
25 Rogers CD,Saxena A,Bronner ME.Sip1 mediates an E-cadherin-to-N-cadherin switch during cranial neural crest EMT.J Cell Bio 2013;203(5):835-847.
26 Chen L,Mu?ozantonia T,Cress WD.Trim28 Contributes to EMT via Regulation of E-Cadherin and N-Cadherin in lung cancer cell lines.Plos One 2014;9(7):e101040.
27 Araki K,Shimura T,Suzuki H,et al.E/N-cadherin switch mediates cancer progression via TGF-β-induced epithelial-to-mesenchymal transition in extrahepatic cholangiocarcinoma.Br J Cancer 2011;105(12):1885-1893.
28 Zhang J,Zhou Y,Yang Y.CCR7 pathway induces epithelial-mesenchymal transition through up-regulation of Snail signaling in gastric cancer.Med Oncol 2015;32(2):1-9.
29 Kurrey NK,Amit K,Bapat SA.Snail and slug are major determinants of ovarian cancer invasiveness at the transcription level.Gynecol Oncol 2005,97(1):155-165.
30 Dinesh B,Sandeep M,Gayatri R,et al.Expression analysis of E-cadherin,Slug and GSK3-βin invasive ductal carcinoma of breast.BMC Cancer 2009;9(1):325.
31 Ricciardi M,Zanotto M,Malpeli G,et al.Epithelial-to-mesenchymal transition(EMT)induced by inflammatory priming elicits mesenchymal stromal cell-like immune-modulatory properties in cancer cells.Br J Cancer2015;112(6):1067-1075.
32 Liu C,Zhang Y,Zhan J,et al.Interleukin-23A is associated with tumor growth in Helicobacter-pylori-related human gastric cancer.Cancer Cell Int 2014;14(1):104.
33 Neuzillet C,Tijeras-Raballand A,Cohen R,et al.Targeting the TGFβpathway for cancer therapy.Pharmacol Ther 2015;147(10):22-31.
34 Heldin CH,Vanlandewijck M,Moustakas A.Regulation of EMT by TGFβin cancer.Febs Letters 2012;586(14):1959-1970.
35 Liu F,Garcia AMG,Meyskens FL.NADPH Oxidase 1overexpression enhances invasion via matrix metalloproteinase-2 and epithelial-mesenchymal transition in melanoma cells.J Invest Dermatol 2012;132(8):2033-2041.
36 Kim ES,Kim MS,Moon A.TGF-beta-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK,but not ERK signaling in MCF10A human breast epithelial cells.Int J Oncol 2004;25(5):1375-1382.
37 Xin N,Hasan M,Li W,et al.Juglans mandshurica Maxim extracts exhibit antitumor activity on HeLa cells in vitro.Mol Med Rep 2014;9(4):1313-1318.
38 Polonik SG,Prokof'Eva NG,Agafonova IG,et al.Antitumor and immunostimulating activity of 5-Hydroxy-1,4-naphthoquinone(Juglone)O-and S-Acetylglycosides.Pharm Che J 2003;37(8):397-398.
39 Fang F,Qin Y,Qi L,et al.Juglone exerts antitumor effect in ovarian cancer cells.Iran J Basic Med Sci 2015;18(6):544-548.
40 Yamazaki T,Tokiwa T.Isofraxidin,a Coumarin component from acanthopanax senticosus,inhibits matrix metalloproteinase-7 expression and cell invasion of human hepatoma cells.Biol Pharm Bull 2010;33(10):1716-1722.
41 Xu R,Kang Q,Ren J,et al.Antitumor molecular mechanism of chlorogenic acid on inducting genes GSK-3βand APC and inhibiting geneβ-catenin.J Anal Methods Chem 2013;2013:951319.
42 Li N,Sun Y N,Zhang L,et al.NF-κB inhibitory activities of triterpenoid glycosides from the stems of Acanthopanax senticosus(Rupr.&Maxim.)Harms.Phytochem Lett 2016;15:210-214.
2 Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015.CA Cancer J Clin 2016;66(2):115-132.
3 Huang L,Wu RL,Xu AM.Epithelial-mesenchymal transition in gastric cancer.Am J Transl Res 2015;7(11):2141-2158.
4 Hartgrink HH,Jansen EP,van Grieken NC,et al.Gastric cancer.Lancet 2009;374(9688):477-490.
5 YE X,Weinberg RA.Epithelial-mesenchymal plasticity:a central regulator of cancer progression.Trends Cell Biol2015;25(11):675-686.
6 Heerboth S,Housman G,Leary M,et al.EMT and tumor metastasis.Clin Translat Med 2015;4(1):6
7 Roussos ET,Keckesova Z,Haley JD,et al.AACR special conference on epithelial-mese nchymal transition and cancer progression and treatment.Cancer Res 2010;70(19):7360-7364.
8 Lander R,Nordin K,Labonne C.The F-box protein Ppa is a common regulator of core EMT factors Twist,Snail,Slug,and Sip1.J Cell Biol 2011;194(1):17-25.
9 Franco-Chuaire ML,Magda Carolina SC,Chuaire-Noack L.Epithelial-mesenchymal transition(EMT):principles and clinical impact in cancer therapy.Invest Clin2013;54(2):186-205.
10 Liu X,Li Z,Song Y,et al.AURKA induces EMT by regulating histone modification through Wnt/β-catenin and PI3K/Akt signaling pathway in gastric cancer.Oncotarget2016;7(22):33152-33164.
11 Li W,Li S,Deng L,et al.Decreased MT1-MMP in gastric cancer suppressed cell migration and invasion via regulating MMPs and EMT.Tumour Biol 2015;36(9):6883-6889.
12 Zohn IE,Li Y,Skolnik EY,et al.p38 and a p38-interacting protein are critical for downregulation of E-cadherin during mouse gastrulation.Cell 2006;125(5):957-969.
13 Krafft E,Lybaert P,Roels E,et al.Transforming growth factor beta 1 activation,storage,and signaling pathways in idiopathic pulmonary fibrosis in dogs.J Vet Intern Med2014;28(6):1666-1675.
14 Strand DW,Liang YY,Yang F,et al.TGF-βinduction of FGF-2 expression in stromal cells requires integrated smad 3 and MAPK pathways.Am J Clin Exp Urol 2014;2(3):239-248.
15 Xu J,Lamouille S,Derynck R.TGF-β-induced epithelial to mesenchymal transition.Cell Research 2009;19(2):156-172.
16 Rees JR,Onwuegbusi BA,Save VE,et al.In vivo and in vitro evidence for transforming growth factor-beta1-mediated epithelial to mesenchymal transition in esophageal adenocarcinoma.Cancer Res 2006;66(19):9583-9590.
17 Hasan TN,Lg B,Shafi G,et al.Anti-proliferative effects of organic extracts from root bark of Juglans Regia L.(RB-JR)on MDA-MB-231 human breast cancer cells:role of Bcl-2/Bax,caspases and Tp53.Asian-Pac.J.Cancer Prev2011;12(2):525-530.
18 Liu JX,Di DL,Wei XN,et al.Cytotoxic diarylheptanoids from the pericarps of walnuts(Juglans regia).Planta Med2008;74(7):754-759.
19 Salimi M,Majd A,Sepahdar Z,et al.Cytotoxicity effects of various Juglans regia(walnut)leaf extracts in human cancer cell lines.Pharm Biol 2012;50(11):1416-1422.
20 Huang L,Zhao H,Huang B,et al.Acanthopanax senticosus:review of botany,chemistry and pharmacology.Pharmazie 2011;66(2):83-97.
21 Herszenyi L,Hritz I,Pregun I,et al.Alterations of glutathione S-transferase and matrix metalloproteinase-9 expressions are early events in esophageal carcinogenesis.World JGastroenterology 2007;13(5):676-682.
22 Guo J,Wang B,Fu Z,et al.Hypoxic microenvironment induces EMT and upgrades stem-like properties of gastric cancer cells.Technol Cancer Res Treat 2016;15(1):60-68.
23 Thiery JP,Acloque H,Huang RYJ,et al.Epithelial-mesenchymal transitions in development and disease.Cell 2014;139(5):871-890.
24 Kalluri R,Weinberg RA.The basics of epithelial-mesenchymal transition.J Clin Invest 2009;119(6):1420-1428.
25 Rogers CD,Saxena A,Bronner ME.Sip1 mediates an E-cadherin-to-N-cadherin switch during cranial neural crest EMT.J Cell Bio 2013;203(5):835-847.
26 Chen L,Mu?ozantonia T,Cress WD.Trim28 Contributes to EMT via Regulation of E-Cadherin and N-Cadherin in lung cancer cell lines.Plos One 2014;9(7):e101040.
27 Araki K,Shimura T,Suzuki H,et al.E/N-cadherin switch mediates cancer progression via TGF-β-induced epithelial-to-mesenchymal transition in extrahepatic cholangiocarcinoma.Br J Cancer 2011;105(12):1885-1893.
28 Zhang J,Zhou Y,Yang Y.CCR7 pathway induces epithelial-mesenchymal transition through up-regulation of Snail signaling in gastric cancer.Med Oncol 2015;32(2):1-9.
29 Kurrey NK,Amit K,Bapat SA.Snail and slug are major determinants of ovarian cancer invasiveness at the transcription level.Gynecol Oncol 2005,97(1):155-165.
30 Dinesh B,Sandeep M,Gayatri R,et al.Expression analysis of E-cadherin,Slug and GSK3-βin invasive ductal carcinoma of breast.BMC Cancer 2009;9(1):325.
31 Ricciardi M,Zanotto M,Malpeli G,et al.Epithelial-to-mesenchymal transition(EMT)induced by inflammatory priming elicits mesenchymal stromal cell-like immune-modulatory properties in cancer cells.Br J Cancer2015;112(6):1067-1075.
32 Liu C,Zhang Y,Zhan J,et al.Interleukin-23A is associated with tumor growth in Helicobacter-pylori-related human gastric cancer.Cancer Cell Int 2014;14(1):104.
33 Neuzillet C,Tijeras-Raballand A,Cohen R,et al.Targeting the TGFβpathway for cancer therapy.Pharmacol Ther 2015;147(10):22-31.
34 Heldin CH,Vanlandewijck M,Moustakas A.Regulation of EMT by TGFβin cancer.Febs Letters 2012;586(14):1959-1970.
35 Liu F,Garcia AMG,Meyskens FL.NADPH Oxidase 1overexpression enhances invasion via matrix metalloproteinase-2 and epithelial-mesenchymal transition in melanoma cells.J Invest Dermatol 2012;132(8):2033-2041.
36 Kim ES,Kim MS,Moon A.TGF-beta-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK,but not ERK signaling in MCF10A human breast epithelial cells.Int J Oncol 2004;25(5):1375-1382.
37 Xin N,Hasan M,Li W,et al.Juglans mandshurica Maxim extracts exhibit antitumor activity on HeLa cells in vitro.Mol Med Rep 2014;9(4):1313-1318.
38 Polonik SG,Prokof'Eva NG,Agafonova IG,et al.Antitumor and immunostimulating activity of 5-Hydroxy-1,4-naphthoquinone(Juglone)O-and S-Acetylglycosides.Pharm Che J 2003;37(8):397-398.
39 Fang F,Qin Y,Qi L,et al.Juglone exerts antitumor effect in ovarian cancer cells.Iran J Basic Med Sci 2015;18(6):544-548.
40 Yamazaki T,Tokiwa T.Isofraxidin,a Coumarin component from acanthopanax senticosus,inhibits matrix metalloproteinase-7 expression and cell invasion of human hepatoma cells.Biol Pharm Bull 2010;33(10):1716-1722.
41 Xu R,Kang Q,Ren J,et al.Antitumor molecular mechanism of chlorogenic acid on inducting genes GSK-3βand APC and inhibiting geneβ-catenin.J Anal Methods Chem 2013;2013:951319.
42 Li N,Sun Y N,Zhang L,et al.NF-κB inhibitory activities of triterpenoid glycosides from the stems of Acanthopanax senticosus(Rupr.&Maxim.)Harms.Phytochem Lett 2016;15:210-214.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |