The Expression Of BDNF, TrkB In The Primary Liver Cancer And Effects Of BDNF On Liver Cancer Cell Line Bel-7402

Objective:The expression of BDNF and TrkB was detected by immunohistochemistry in 8 cases of normal liver tissue,48 cases of liver cancer and their para-cancer tissue respectively to analyze the relationship between BDNF,TrkB expression and the clinical pathological features and recurrence,survival rate.Methods:Immunohistochemistry was performed to investigate the expression of BDNF and TrkB in 8 cases of normal liver tissue and 48 cases of liver cancer and its para-cacner tissue.Results:There was no BDNF and TrkB protein expression observed in normal liver tissue, while expressed in liver cancer,with the rate of 60.4%(29/48)and 52.1%(25/48),which was higher than their para-cancer tissues(P<0.05)respectively.Significant relations with Edmondson Grade in BDNF and TrkB positive group and capsule formation in BDNF positive group and multiple nodules of tumor in TrkB positive group were observed respectiverly(P<0.05).There was positive correlation between BDNF and TrkB(r=0.332,P<0.05).and both of which showed significant relations with the two year's recurrence rate (P<0.05)and survival time(P<0.01);And they were not associated with no significant relation with age,gender,HBsAg,AFP,cirrosis and the size of tumor in patients with liver cancer.Conclusion:(1)The protein expression of BDNF and TrkB are higher in liver cancer than in their para-liver cancer. Which were not expressed in normal liver tissue.(2)The positive rates of BDNF and TrkB are tightly associated with Edmondson Grade together and capsule formation,multiple nodules of tumor in each positive group respectively. There was positive correlation between BDNF and TrkB(r=0.332, P<0.05).(3)the high expression of BDNF and TrkB maybe signify the earlier recurrence and poor prognosis.Objective:To investigate the effects of exogenous BDNF on biological behavior and expression of Bcl-2 in liver cancer cell line Bel-7402.Methods:control group, low concentration group, middle concentration group and high concentration group were cultured with RPMI 1640 containing Ong/ml 25ng/ml,50ng/ml and 100ng/ml BDNF respectively.Forethermore,the normal liver cell line L-02 was used as normal control group in the mobility,invasion and MTT,RT-PCR assay (1)Cell proliferation of Bel-7402 was estimated by MTT assay;(2) by the way of establishing suspension culture model by culture plate treated with Poly-HEMA, the cell anoikis rate was analyzed by flow cytometry(FCM);(3)Adhesion ability was analyzed by adhesion assay;(4) the mobility ability was detected by scrape migration assay;(5)Invasion ability was analyzed by Transwell chamber assay;(6) Expression level of mRNA and protein of BDNF,TrkB and Bcl-2 were investigated by RT-PCR and Western Blot respectively.Results:Promoting cell proliferation and the ability of adhesion were observed in MTT and Adhesion assay (P<0.01),and the ability to resist anoikis enhanced by BDNF showed in Flow cytometry (FCM) (P<0.01),and transwell chamber assay and scrape migration assay also showed that BDNF could enhance the ability of invasion and mobility of Bel-7402 in no concentration-dependent manner way (P<0.01). Expression of Bcl-2 mRNA and its protein of in Bel-7402 was up-regulated by BDNF showed in RT-PCR and Western Blot (p<0.05), and there was no significant difference among different groups.Conclusion:(1)BDNF and TrkB protein are expressed in liver cancer Bel-7402 cell line;(2) Exogenous BDNF can promote cell proliferation,as well as enhance the ability to resist anoikis and the ability of adhesion,mobility and invasion in liver cancer Bel-7402 cell line;(3)Exogenous BDNF can up-regulate the expression of Bcl-2 gene and protein in Bel-7402 cell.Objective:To investigate anti-apoptosis effects of BDNF induced by 5-Fu in vitro and the effects of exogenous BDNF on the growth of transplanted tumor of Bel-7402 in nude mice.Methods(1)assay in vitro:ontrol group,A(5-Fu) group and B (5-fu+BDNF)group were established to detect anti-apoptosis effects of BDNF induced by 5-Fu on Bel-7402 cell line by MTT assay,clone formation assay and FCM respectively.(2)assay in vivo:for establishing mouse transplanted tumour model,0.9% sodium chloride solution and BDNF were injected subcutaneously into nude mice of control group and BDNF group respectively for the purpose of establishing mouse transplanted tumour model,and growth rate of tumor(including size and weight) was detected, the histological morphous was observed by HE staining, the expression of BDNF and TrkB was detected by immunohistochemistry, finally the expression level of mRNA and protein of Bcl-2 was investigated by RT-PCR and Western Blot. Results:(1)MTT assay,clone formation assay and FCM showed BDNF could inhabit the apoptosis effects induced by 5-Fu in Bel-7402 cell line.(2)Transplanted human liver cancer were developed in all nude mice,growth of tumor was significantly promoted by BDNF. The tumor volume and weight in BDNF group were significant more than control group (P<0.05).(3)HE staining showed typical histological morphous of liver cancer of transplantation tumor tissue.(3)Immunohistochemical staining showed:there were expressions of BDNF and TrkB protein in transplantation tumor tissue.(4)RT-PCR and Western Blot showed that BDNF could up-regulate the expression of mRNA and protein of Bcl-2 in transplantation tumor(P<0.05).Conclusion:(1)Exogenous BDNF has the anti-apoptosis effects induced by 5-Fu in Bel-7402 cell line.(2)Exogenous BDNF can promote the growth of transplantation tumor. (3)There are expression of BDNF and TrkB in transplanted tumor tissue. (4)Exogenous BDNF can up-regulate the expression of Bcl-2 gene in transplanted tumor...