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

为探讨胆钙素对胰腺癌细胞增殖抑制作用的差异，将胰腺癌细胞株Suit-2、S2-007、S2-013、S2-020、S2-028与不同浓度的胆钙素共同孵育，用MIT法检测细胞增殖情况。定量RT-PCT法检测维生素D受体(VDR)的信使RNA水平。结果表明，不同浓度的胆钙醇对细胞增殖的抑制作用不同，以S2-028最强，S2-007和S2-013中等，S2-002弱。定量逆转录聚合酶链式反应显示，VDR基因在S-028细胞中有较高的组成基因表达，且胆钙化醇对VDR基因的诱导作用强于其他细胞。为了研究与细胞增殖和转移相关的基因表达，对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、TTiE、TEK、AU、PLAUR、PLAUR、GGAFC、GAIT2、IT3、IT4、ITGA5、IT6、CTNA1、HBV、HT1、GBGA3、GBIT5、CBS、HCIT3、GB1、GBGAFB3、TGFB1、TGFB2、TGFB3、TGFBR3、TEK、AU、PLAUR、PLAUR、GGAFC、GAIT2、IT3、IT4、ITGA5、IT6、CTNA1、HBV、HT1、GBGA3、GBIT5、TGFB1、CBS、HCB3DNMT1、DNMT3A、HDAC1、HDAC2、HDAC3、HDAC4、HDAC5、HDAC6、MAT1A、MAT2A、MAT2B、MBD2、MBD3、MBD4、MECP2、MTHFR、MTR、CTSL、ANGPT2。结果表明，EGFR、MMP1、MMP3、MMP7、TGFB2、TGFBR3、PLAU、ITGA2和ANGPT2等9个基因的mRNA表达水平高于其他亚系。S2-028的基因表达水平高于其他亚系。

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.