内皮素3对恶性黑素瘤A375细胞上皮基质转化的影响

发布时间：2018-05-02 23:28

本文选题：黑素瘤 + 内皮素-3　；参考：《华中科技大学》2010年硕士论文

【摘要】：[目的]研究内皮素-3(Endothelin 3, ET-3)对人恶性黑素瘤A375细胞上皮基质转化(Epithelial-to-Mesenchymal Transition, EMT)的影响。 [方法]体外培养A375细胞株,分别设立不同干预组,A:空白对照组;B:加入ET-3(100nmol/l);C:加入ETRB阻断剂BQ788组( 100nmol/LET3+100μmol/LBQ788 );采用Transwell小室检测细胞的转移,细胞爬片技术检测细胞形态的变化,采用Realtime-PCR和Western Blot检测上皮基质转化相关分子E-cadherin、vimentin及转录因子(Twist、Slug)的表达情况。 [结果]各干预条件中:1)ET-3可以促进黑素瘤A375细胞的转移,ETRB阻断剂BQ788可阻断该效应;2) ET-3可以促进黑素瘤A375细胞从上皮型向成纤维细胞样形态转变;ET3促进黑素瘤A375细胞上皮基质转化标志分子E-cadherin表达下调, Vimentin表达上调,而ETRB阻断剂BQ788可阻断这种效应;3) ET-3可以促进上皮基质转化相关转录因子Twist及Slug在蛋白和mRNA水平上表达上调。 [结论] ET-3/ETRB通路通过调节上皮基质转化相关分子E-cadherin、Vimentin及转录因子(Twist、Slug)促进黑素瘤A375细胞上皮基质转化。
[Abstract]:[objective] to study the effect of endothelin 3 (et 3) on epithelial matrix transformation of human malignant melanoma A375 cell line Epithelial-to-mesenchymal transition (EMTT). [methods] A375 cell lines were cultured in vitro. Different intervention groups were set up respectively: control group B: add ET-3N 100nmol / L ETRB inhibitor BQ788 group (100nmol/LET3 100 渭 mol/LBQ788), cell metastasis was detected by Transwell chamber and cell morphology was detected by cell climbing technique. Realtime-PCR and Western Blot were used to detect the expression of E-cadherin vimentin and Twistus Slug. [results] among the different intervention conditions, the effect was blocked by BQ788, the inhibitor of ETRB, which could promote the metastasis of A375 cells.) ET-3 could promote the transformation of A375 cells from epithelial to fibroblast-like morphology. Et 3 could promote the fine morphology of A375 melanoma cells. The expression of E-cadherin was down-regulated and the expression of Vimentin was up-regulated. BQ788, a ETRB blocker, could block this effect. ET-3 could increase the expression of Twist and Slug at the level of protein and mRNA. [conclusion] the ET-3/ETRB pathway promotes the epithelial matrix transformation of A375 cells by regulating the epithelial matrix transformation related molecules E-cadherinus vimentin and transcriptional factor Twisten Slug.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.5

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