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Difference in the anti-proliferative effect by cholecalciferol between sublines of a pancreatic cancer cell lines and various genes expressions examined by cDNA array

胆钙化醇对胰腺癌细胞系亚系间抗增殖作用的差异及cDNA芯片检测各基因表达

基本信息

批准号：
15591416
负责人：
ETO Tadaaki
金额：
$ 1.92万
依托单位：
University of Miyazaki (2004)宮崎医科大学 (2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591416/
关键词：
pancreatic carcinoma MTT assay cholecalciferol vitamine D receptor cDNA array cDNA array MTT assa 増殖・転移抑制

项目摘要

To examine the difference in the anti-proliferative effect by cholecalciferol between pancreatic cancer cells by cholecalciferol, four sublines of a pancreatic cancer cell line SUIT-2,S2-007,S2-013,S2-020, and S2-028 were incubated at varied concentrations of cholecalciferol and the cellular proliferation were assessed by MIT assay. Messenger RNA levels of vitamin D receptor (VDR) were also measured by quantitative RT-PCT. As a result cholecalciferol inhibited the cellular proliferation depending on the concentrations (10^<-6> M,10^<-7> M,10^<-8> M), most strongly in S2-028, moderately in S2-007 and S2-013, and weekly in S2-002. Quantitative reverse transcriptase polymerase chain reaction showed higher constitutional gene expression of VDR mRNA in S-028 and greater induction by cholecalciferol than those in the other sublimes. To investigate the gene expressions, related to cellular proliferation and metastasis in these sublines, cDNA array analysis was performed in genes as follows : ACTB,AMFR,FGF2,CTSB,PECAM1,CD34,CDH1,EGFR,ETS1,FLT1,FLT4,FOS,KDR,MMP1,MMP2,MMP3,MMP7,MMP9,MET,MMP14,MMP16,MYC,RELA,NME1,NME2,SERPINE1,SERPINB2,TFRC,TGFA,TGFB1,TGFB2,TGFB3,TGFBR3,TIE,TEK,PLAU,PLAUR,VEGF,VEGFC,ITGA2,ITGA3,ITGA4,ITGA5,ITGA6,CTNNA1,ITGAV,THBS1,HTGB1,ITGB3,ITGB5,CTNNB1,AHCY,CBS,DNMT1,DNMT3A,HDAC1,HDAC2,HDAC3,HDAC4,HDAC5,HDAC6,MAT1A,MAT2A,MAT2B,MBD2,MBD3,MBD4,MECP2,MTHFR,MTR,CTSL,ANGPT2. As a result mRNA expression levels were higher than those of the other sublines in nine genes such as EGFR,MMP1,MMP3,MMP7,TGFB2,TGFBR3,PLAU,ITGA2, and ANGPT2. S2-028 showed higher gene expression than those in the other sublines.

为了检查胆钙化醇在胰腺癌细胞之间的胆钙化醇抗增殖作用的差异，将胰腺癌细胞系SUIT-2、S2-007、S2-013、S2-020和S2-028的四个亚系在不同浓度的胆钙化醇下孵育，并通过MIT测定评估细胞增殖。还通过定量RT-PCT测量维生素D受体（VDR）的信使RNA水平。因此，胆钙化醇根据浓度（10 μ <-6>M、10 μ <-7>M、10 μ <-8>M）抑制细胞增殖，在S2-028中最强，在S2-007和S2-013中中等，在S2-002中每周一次。定量逆转录聚合酶链反应显示，S-028中VDR mRNA的组成基因表达高于其他升华物，且胆钙化醇的诱导作用更强。为了研究这些亚系中与细胞增殖和转移相关的基因表达，在如下基因中进行cDNA阵列分析：ACTB、AMFR、FGF2、CTSB、PECAM 1、CD 34、CDH 1、EGFR、ETS1、FLT 1、FLT 4、FOS、KDR、MMP 1、MMP 2、MMP 3、MMP 7、MMP 9、MET、MMP 14、MMP 16、MYC、RELA、NME 1、NME 2、SERPINE1、SERPINB 2、TFRC、TGFA、TGFB1、TGFB2、TGFB3、TIE、TEK、PLAU、PLAUR、VEGF、VEGFC、ITGA 2、ITGA 3、ITGA 4、ITGA 5、ITGA 6、CTNNA 1、ITGV、THBS1、HTGB 1、ITGB 3、ITGB 5、CTNNB 1、AHCY、CBS、DNMT 1、DNMT 3A、HDAC 1、HDAC 2、HDAC 3、HDAC 4、HDAC 5、HDAC 6、MAT 1A、MAT 2A、MAT 2B、MBD2、MBD3、MBD4、MECP 2、MTHFR、MTR、CTSL、ANGPT 2。结果，EGFR、MMP 1、MMP 3、MMP 7、TGFB 2、TGFBR 3、PLAU、ITGA 2和ANGPT 2等9个基因的mRNA表达水平高于其他亚系。S2-028的基因表达量明显高于其它亚系。