Searching for vasculogenic mimicry-associated differentially expressed genes in human hepatocellular carcinoma cell line MHCC97 using functional grouping cDNA microarray
10.3760/cma.j.issn.1673-9752.2011.04.011
- VernacularTitle:基因芯片筛选人肝癌细胞株MHCC97中血管生成拟态相关基因
- Author:
Wenbin LIU
;
Geliang XU
;
Weidong JIA
;
Jiansheng LI
;
Jinliang MA
;
Yongsheng GE
;
Ye HU
;
Hao CHEN
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Vasculogenic mimicry;
Three-dimensional culture;
Gene chip
- From:
Chinese Journal of Digestive Surgery
2011;10(4):278-281
- CountryChina
- Language:Chinese
-
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.
