Effects Of Lupeol On Expression Of Wnt System In Human Hepatoma Cell Line HepG2

Objective: Hepatocellular carcinoma is the world’s fifth malignant tumors,ranking the third leading cause of death, the mortality of liver cancer is aftergastric cancer, esophageal cancer.The liver cancer incidence rate in China is the highest in the world,because China is a big country of hepatitis. Liver cancer has become one ofthe main diseases that threaten human health. Liver cancer’s early symptomsare not obvious, mainly for advanced liver pain, fatigue, weight loss, jaundice,ascites and other symptoms. When patients with typical clinical manifestationssuch as in the late fall, losing the chance of operation, the prognosis is verypoor. The natural survival is not more than six months. At present in thetreatment of HCC, surgical operation is still the first choice as the besttreatment method recognized nationally and internationally, but liver resectionfor hepatocellular carcinoma has a high propensity for local recurrence,studies have found, a lot of primary liver cancer after radical resection has ahigh recurrence rate--70%, although there are a lot of patients afterconventional chemotherapy, the high recurrence rate seriously affected theoperation and the effect of chemotherapy in patients with hepatocellularcarcinoma, many lost their precious lives, how to predict liver cancer’s highrecurrence rate, and the inhibition of recurrence effectively is the key toimprove the treatment of liver cancer.The pathogenesis of hepatocellular carcinoma is complex, multi factor,multi genes, multi stages.Involved in the cell signal transduction pathways,the Wnt/β-catenin transduction pathway plays a vital role in the transductionpathway. In the path through the beta catenin (β-catenin) accumulated in thenucleus and activate the target genes. The new study shows that-cateninprotein plays an important role in promoting tumor development process, Wnt/ β-catenin transduction pathway is not activated in normal cells. In normalcells, β-catenin is only as a cytoskeletal protein adhesion, to maintain thepeer cells and prevent cell migration,it plays a role in cell membrane, onlywhen the Wnt signal molecular and cell membrane specific receptor bindingthey activate the intracellular protein, the β-catenin is avoid beingphosphorylated by degradation fate, β-catenin can accumulate in thecytoplasm. When beta catenin concentration reaches a certain level, it cantransfer to the nucleus, it can combine with TCF,the target gene c-MYC andcyclin can be transcription,Wnt path way is open.If there is no Wnt signal,β-catenin was decomposed,Wnt path way is closed.To find an efficient and low toxicity drug become people’stoppriority.Lupeol widely exists in a variety of fruits, vegetables and herbs andother plants, it has a strong effect of anti-inflammatory, anti gene mutation andmalaria. Recent studies suggest that lupeol has anti-cancer effects, in additionto human toxicity.Method: The conventional culture of human hepatocellular carcinoma cellline, HepG2were divided into control group and test group. The control groupcells were cultured, cells in experimental groups were applied lupeolintervention, a concentration gradient and time gradient. Use reversetranscription polymerase chain reaction (RT-PCR) technique, measuring thechanges of expression of Wnt3and β-catenin mRNA, with β-actin asinternal reference standard, the amplified products of value DNAelectrophoresis band optical density analysis by gel scanner, Wnt3’s ratio toβ-actin as the expression levels of the parameters for the Wnt3mRNA,; theβ-catenin’s ratio to β-actin as a parameter β-catenin expression level ofmRNA. With different concentrations of lupeol (0,15,25,35μ mol/L)treatment of HepG2cells, the concentration of lupeol Western-Blot method (0,15,25,35μ mol/L) HepG2cells were treated with24h、48h、72h, theexpression changes of Wnt3, β-catenin protein in HepG2cells. All data wereexpressed with±s, using SPSS13.0statistical analysis software and singlefactor analysis of variance (ANOVA) and t test. P<0.05means significant difference.Result: Lupeol’s effect of different concentration for different time, theexpression of HepG2cell Wnt3and β-catenin mRNA compared with thecontrol group decreased gradually, and lupeol concentration of15-35μ mol/Lrange is concentration dependent and time dependent (P<0.05).Lupeol’s effect of different concentration of different time also has a inhibitoryeffect on HepG2cells in Wnt3and β-catenin protein, the expression of itschanges agrees with the trends of Wnt3and β-catenin mRNA, and also has atime difference effect (P<0.05).Conclusion: The expression of lupeol can inhibit Wnt3and-catenin βhepatocellular carcinoma in nucleic acid and protein level.