紫花牡荆素干预食管鳞状细胞癌细胞转移的分子机制
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摘要
目的研究紫花牡荆素对人食管鳞状细胞癌(食管鳞癌)细胞恶性转移能力的抑制效应并探讨其分子机制。方法人食管鳞癌细胞株EC9706与EC109细胞经紫花牡荆素处理后,以MTT比色法测定细胞增殖能力,侵袭与迁移实验检测细胞转移能力,Western blot检测转移相关蛋白表达变化。结果不同浓度紫花牡荆素处理可显著抑制EC9706与EC109细胞的增殖、侵袭与迁移能力,同未加药组比较差异均有统计学意义(P<0.05);紫花牡荆素处理下调兔抗人血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶2(matrix metalloproteinase2,MMP2)与基质金属蛋白酶9(matrix metalloproteinase9,MMP9)表达水平,上调上皮细胞钙黏蛋白(E-cadherin)、周期素依赖性激酶10(cyclin-dependent kinase 10,CDK10)与兔抗人受体O型蛋白质酪氨酸磷酸酶(protein tyrosine phosphatase receptor type O,PTPRO)表达。结论紫花牡荆素对食管鳞癌细胞恶性转移能力有明显的抑制作用,下调VEGF、MMP2与MMP9表达,上调E-cadherin、CDK10与PTPRO可能是其抑制转移的分子机制。
Objective To investigate the depressive effect of Casticin on malignant metastasis of human esophagus squamous cell carcinoma( ESCC) cells and to analyze its molecular mechanisms. Methods The EC9706 and EC109 cells in ESCC cell line were treated with Casticin,and then the thiazolyl blue( MTT) colorimetry was used to detect cell abilities of proliferation,invasion and migration,and related protein expressions were detected using Western blot method. Results EC9706 and EC109 cells abilities of proliferation,invasion and migration were significantly inhibited after different concentrations of Casticin treatment,and the differences were statistically significant compared with those without Casticin treatment( P < 0. 05); after the Casticin treatment,the expressions of vascular endothelial growth factor( VEGF),matrix metalloproteinase-2( MMP2) and matrix metalloproteinase-9( MMP9) were decreased,while the expressions of epithelium-cadherin( E-cadherin),cyclin dependent kinase10( CDK10) and protein tyrosine phosphatase receptor type O( PTPRO) were increased. Conclusion Casticin may significantly inhibit malignant metastasis of ESCC cells,and down-regulation of VEGF,MMP2 and MMP9 expressions and up-regulation of E-cadherin,CDK10 and PTPRO expressions. Its may be the molecular mechanisms to inhibit its transfer.
Objective To investigate the depressive effect of Casticin on malignant metastasis of human esophagus squamous cell carcinoma( ESCC) cells and to analyze its molecular mechanisms. Methods The EC9706 and EC109 cells in ESCC cell line were treated with Casticin,and then the thiazolyl blue( MTT) colorimetry was used to detect cell abilities of proliferation,invasion and migration,and related protein expressions were detected using Western blot method. Results EC9706 and EC109 cells abilities of proliferation,invasion and migration were significantly inhibited after different concentrations of Casticin treatment,and the differences were statistically significant compared with those without Casticin treatment( P < 0. 05); after the Casticin treatment,the expressions of vascular endothelial growth factor( VEGF),matrix metalloproteinase-2( MMP2) and matrix metalloproteinase-9( MMP9) were decreased,while the expressions of epithelium-cadherin( E-cadherin),cyclin dependent kinase10( CDK10) and protein tyrosine phosphatase receptor type O( PTPRO) were increased. Conclusion Casticin may significantly inhibit malignant metastasis of ESCC cells,and down-regulation of VEGF,MMP2 and MMP9 expressions and up-regulation of E-cadherin,CDK10 and PTPRO expressions. Its may be the molecular mechanisms to inhibit its transfer.
引文
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[2]黄继红,游颜杰,郑坚.紫花牡荆素诱导食管癌细胞凋亡的分子机制[J].中国现代医生,2013,2(4):197-199.
[3]Chen D,Cao J,Tian L,et al.Induction of apoptosis by casticin in cervical cancer cells through reactive oxygen species-mediated mitochondrial signaling pathways[J].Oncol Rep,2011,26(8):1287-1294.
[4]Zhou Y,Tian L,Long L,et al.Casticin potentiates TRAIL-induced apoptosis of gastric cancer cells through endoplasmic reticulum stress[J].PLOS One,2013,8(3):e58855.
[5]王奕鸿,刘济生,盛伟华,等.RGD修饰的腺病毒介导的ING4对人鼻咽癌细胞裸鼠移植瘤的影响[J].解放军医药杂志,2014,26(4):50-54.
[6]王光茂,原银萍,赵翠秀,等.结肠癌组织MMP2和MMP9及VEGF表达临床意义的研究[J].中国医师杂志,2011,18(16):1267-1269.
[7]Clarke J M,Hurwitz H I.Targeted inhibition of VEGF receptor 2:an update on ramucirumab[J].Expert Opin Biol Ther,2013,13(8):1187-1196.
[8]You Y,Liu J,Wang Z,et al.The enhancement of radiosensitivity in human esophageal squamous cell carcinoma cells by zoledronic acid and its potential mechanism[J].Cytotechnology,2014,66(1):17-25.
[9]Domigan C K,Warren C M,Antanesian V,et al.Autocrine VEGF maintains endothelial survival through regulation of metabolism and autophagy[J].Cell Sci,2015,128(12):2236-2248.
[10]Wang L,Ji S,Cheng Z.Association between polymorphisms in vascular endothelial growth factor gene and response to chemotherapies in colorectal cance:A meta-analysis[J].PLo S One,2015,10(5):e0126619.
[11]Ran Y,Wu S,You Y.Demethylation of E-cadherin gene in nasopharyngeal carcinoma could serve as a potential therapeutic strategy[J].Biochem,2011,149(1):49-54.
[12]Jin Y,Shenoy A K,Doernberg S,et al.FBXO11 promotes ubiquitination of the Snail family of transcription factors in cancer progression and epidermal development[J].Cancer Lett,2015,362(1):70-82.
[13]Tiwari I,Yoon M H,Park B J,et al.Hepatitis C virus core protein induces epithelial-mesenchymal transition in human hepatocytes by upregulating E12/E47 levels[J].Cancer Lett,2015,362(1):131-138.
[14]Verduzco D,Lloyd M,Xu L,et al.Intermittent hypoxia selects for genotypes and phenotypes that increase survival invasion and therapy resistance.[J].PLo S One,2015,10(3):e0120958.
[15]You Y,Chen Y,Zheng X,et al.Aberrant methylation of the PTPRO gene in peripheral blood as a potential biomarker in esophageal squamous cell carcinoma patients[J].Cancer Lett,2012,315(2):138-144.
[16]Zhang W,Hou J,Wang X,et al.PTPRO-mediated autophagy prevents hepatosteatosis and tumorigenesis[J].Oncotarget,2015,6(11):9420-9433.
[17]Zhang X,Tan Z,Wang Y,et al.Protein tyrosine phosphatase receptor type O expression in the tumor niche correlates with reduced tumor growth,angiogenesis,circulating tumor cells and metastasis of breast cancer[J].Oncol Rep,2015,33(4):1908-1914.
[18]You Y,Yang W,Wang Z,et al.Promoter hypermethylation contributes to the frequent suppression of the CDK10gene in human nasopharyngeal carcinomas[J].Cell Oncol(Dordr),2013,36(4):323-331.
[19]Zhong X Y,Xu X X,Yu J H,et al.Clinical and biological significance of Cdk10 in hepatocellular carcinoma[J].Gene,2012,498(1):68-74.
[20]Khanal P,Yun H J,Lim S C,et al.Proyl isomerase Pin1 facilitates ubiquitin-mediated degradation of cyclindependent kinase 10 to induce tamoxifen resistance in breast cancer cells[J].Oncogene,2012,31(34):3845-3856.