Relation Between Wif-1Promot Methylation And Hepatocellular Carcinoma

WNT signal widely involves the growth of organisms and tumordevelopment. At present, many studies show that WNT pathway canopen,which dues to its downstream target genes mutation or deletion, suchas GSK-3beta, C-myc, Cycline D1, Axin, ngn1. However, it is suggestedthat the upstream signals of WNT signaling pathways, such as WNT proteinand its inhibitors (Wif-1, WNT2, and WNT3) can be find alterations andepigenetic changes relatively. The expression of WNT proteins weresignificantly reduced In Hepatocellular carcinoma compared to in normalliver tissues. It is speculated that the development of Hepatocellularcarcinoma is closely related to WNT signal pathway. In addition, β-cateninheterogeneous expression, the down streams of WNT signaling pathway,plays a critical factor. At the same time, many studies show that WNTpathway downstream target genes such as C-myc and CyclineD1alsoincreased. There are rare reports confirm which is much more relativecorrelation between β-catenin heterogeneous expression and Wnt-signalingupstream and downstream genes.Wif-1(Wnt inhibitory factor-1) is one of the WNT signal inhibitor, which combine to block WNT protein. But theunderlying mechanism of down-regulation of Wif-1and its relativebiological influences on hepatocellular carcinoma.Objective:（1）To detect the expression of Wif-1, C-myc, and theCycling-D1in L0and HepG2and to explore the correction between themand the expression of β-catenin heterogeneous（.2）To tset whether promotermethylation of Wif-1inhibits the gene expression.（3）To explore therelation between Wif-1re-expression and procaine,and to explore thepotential machinery of5-aza-2’-deoxycytidine and procaine by analyzingtheir combination effect on HepG2.（4）To explore the relationship betweenpromoter methylation of Wif-1and clinical pathology of hepatocellularcarcinoma for the purpose of demonstrating if Wif-1can be used as amolecular for the diagnosis and prognosis of HCC.Methods:（1）Immunohistochemistry and In Situ Hybridization wereused to detect the expression of Wif-1, C-myc and the Cycling-D1on35cases of HCC patients, at the same time35cases of normal ascontrols.(2)Procaine and5-aza-dc drugs were applied to assess theexpression of wif-1, by RT-PCR, nested methylation specific PCR andWestern Blot.(3)RT-PCR, MTT and Western blot were performed to detectthe promoter demethylation changes after procaine and cisplatin In vitro, andprocaine with cisplatin were employed to detect the changes of thesusceptibility of the chemotherapy drugs. Results:(1)Wif-1, C-myc and Cycling-D1expression pattern ofHepatocellular carcinoma and normal liver were identical: IMM and ISHshowed that the Pearson of Wif-1in Hepatocellular carcinoma tissue andnormal liver were r IMM=0.53and r ISH=0.50, of C-myc were rIMM=0.48and r ISH=0.48and of Cycling-D1were rIMM=0.47and rISH=0.44.(p <0.05).(2)Wif-1methylation rates reduced with Procaine or5-aza-dcinterference(14.41%in the experimental group P,29.29%in the controlgroup).(3)There was no Wif-1mRNA and protein expression whenCisplatin interference, but recover the expression when it compared withProcaine (the significant difference (P>0.05).)Conclusions:(1)Upstream played a greater factor with β-cateninheterogeneous expression in Wnt signaling pathway than the downstreamtarget genes such as C-myc, Wif-1, and Cycling-D1.(2)Wif-1can be used asan important molecular marker in the diagnosis of Hepatocellular carcinoma(HCC).(3)The promoter methylation status of Wif-1was significantlyassociated with Procaine than5-aza-dc off a.(4)In Wnt signaling pathway ofHepatocellular carcinoma the susceptibility can be enhanced by cisplatin andprocaine together.