目的采用细胞外基质与粘连分子Real-time PCR基因芯片筛选高、低转移潜能人肝癌HCCLM3、MHCC97-L细胞系差异表达的血管生成拟态基因,探讨血管生成拟态的机制。方法建立HCCLM3、MHCC97-L三维培养体系,倒置显微镜下观察其血管样结构形成情况。细胞培养24 h后,采用细胞外基质与粘连分子Real-timePCR基因芯片对HCCLM3、MHCC97-L的表达基因进行筛选。结果培养24 h,HCCLM3形成的血管样结构较MHCC97-L的长(P〈0.01)。筛选出20个差异基因,其中CD44、COL6A1、COL4A2、ECM1、ITGA1、ITGA3、MMP1、MMP2、SPP1、TNC、COL6A2、MMP7和MMP10共13个基因在HCCLM3中的表达较MHCC97-L明显上调,CTNND2、COL12A1、COL14A1、ITGAL、TIMP3、MMP16和VTN共7个基因明显下调。结论高、低转移潜能人肝癌HCCLM3、MHCC97-L细胞系体外形成血管生成拟态有差异,其原因可能与HCCLM3差异表达某些细胞外基质和粘连分子相关基因有关。
Objective The extracellular matrix and adhesion molecules Real-time PCR array was used to screen the vasculogenic mimicry-associated gene profiles of human hepatocellular carcinoma cell lines HCCLM3 and MHCC97-L with high and low metastatic potentials respectively.Methods Three-dimensional cell culture system of HCCLM3 and MH-CC97-L was established to observe tubelike structures by inverted microscope.Gene expression profiles of HCCLM3 and MH-CC97-L were detected by extracellular matrix and adhesion molecules Real-time PCR array.Results Tubelike structures were observed longer in HCCLM3 than that in MHCC97-L after three-dimensional culture for 24 h(P〈0.01).Among a total of 20 genes identified,13 genes including CD44,COL6A1,COL4A2,ECM1,ITGA1,ITGA3,MMP1,MMP2,SPP1,TNC,COL6A2,MMP7 and MMP10 were up-regulated and 7 genes including CTNND2,COL12A1,COL14A1,ITGAL,TIMP3,MMP16 and VTN were down-regulated in HCCLM3,compared with MHCC97-L.Conclusion HCCLM3 with high metastatic potentials has stronger ability to form vasculogenic mimicry than MHCC97-L with low metastatic potentials and this perhaps correlates with the differential expression of certain extracellular matrix and adhesion molecules-related genes in HCCLM3.