The effects of stbm/vangl (strabismus/van gogh like) gene expression on cell behaviour in hepatocellular cancer cell lines

SummaryThe effects of STBM/VANGL (Strabismus/Van Gogh like) gene expression on cellbehaviour in hepatocellular cancer cell linesG. Ozan ÇetinDokuz Eylül University, Faculty of medicine, Department of Medical biology andGenetics, İzmirHepatocellular cancer (HCC) is the fifth most common cancer type and has noeffective treatment so far.Van Gogh like 1( Vangl1) protein, which is a transmembrane protein takingplace on Wnt planar cell polarity pathway, is important for embryonic cells to gainplanar cell polarity during developmental process and is also responsible fromconvergent extension. In adults, it is expressed spesifically in testis and ovarium as wellas in brain and prostat.VANGL1 expression has been reported in several human cancer cell linesincluding HCC. Our aim in this study was to investigate the behavior of HCC cellwhose VANGL1 gene was suppressed by siRNA method.The existence of VANGL1 expression in HCC cell lines was shown by RT-PCRand real time PCR methods. Internet based software program was used to define thetarget siRNA area spesific for VANGL1 gene. DNA line which transcribes the hairpinsiRNA for this target was designed. SiRNA insert was added to pSilencer 4.1 CMV Neoplasmid, later which was used to transfect HepG2 cells. The colonies having suppressedVANGL1 gene were examined by RT-PCR method. Suppression of the gene expressionwas proven by using western-blot method in assessing protein lisates provided by thecolonies. Motility and invasion of the cells having suppressed VANGL1 were assessedby Boyden chamber assay while proliferation analysis was assessed by flow cytometry.There was no difference in the motility of the HCC cells having suppressedVANGL1 where as the invasion capacity was decreased by 3 times.??. In proliferationanalysis, there was no difference in S phase cell ratio between VANGL1 suppressedcells and parental cells.As a conclusion, VANGL1 gene has an effect on cell invasion rather than cellmotility. Further investigations are needed to understand the mechanism of this effect.Key Words: Wnt, VANGL1, hepatocellular cancer, motility, invasion,proliferation