Searching for vasculogenic mimicry-associated differentially expressed genes in human hepatocellular carcinoma cell line MHCC97 using functional grouping cDNA microarray / 中华消化外科杂志

ABSTRACT

Objective To observe vasculogenic mimicry formation difference in human hepatocellular carcinoma cell lines MHCC97-H/L with different metastatic potentials under three-dimensional culture condition,and to study extracellular matrix and adhesion molecules-related mechanism of vasculogenic mimicry.Methods Three-dimensional cell culture system of MHCC97-H/L was established to observe tubelike structures by inverted microscope.Human umbilical vein endothelial cell(HUVEC),human hepatocellular carcinoma cell line Hep3B with non-metastatic potentials and normal human hepatic cell line HL-7702 were taken as control.Gene expression profiles of MHCC97-H and MHCC97-L were detected by Oligo extracellular matrix and adhesion molecules microarray analysis.Two differentially expressed genes were verified with semi-quantitative reverse transcriptionpolymerase chain reaction(RT-PCR)and Western blot.All data were analyzed using two sample t test.Results After a 24-hour three-dimensional culture,the length of tubelike structures was(474 ± 16)mm/cm2 in MHCC97-H,which were significantly longer than(320 ±41)mm/cm2 in MHCC97-L(t =6.119,P ＜0.05).Hep3B and HL-7702 could not form tubelike structures.Compared with MHCC97-L,the expression of 7 genes were up-regulated and the expression of 3 genes were down-regulated among a total of 113 angiogenesis genes in MHCC97-H.Two of the genes with differential expressions including tenascin C and extracellular matrix protein 1 were further validated by RT-PCR and Western blot.Conclusion MHCC97-H has stronger ability to form vasculogenic mimicry than MHCC97-L,and this perhaps correlates with the differential expression of certain extracellular matrix and adhesion molecules-related genes in MHCC97-H.