The Effect Of TGF-β1siRNA On Bladder Tumor Angiogenesis

Bladder tumor is one of the most common tumor in urology，itis also a disease that directly threat to survival of patients.Its biologic behavioris complicated,mainly for them: recurrence, multiple, infiltration and metastasis,etc.The90%of bladder tumor is urothelial carcinoma,it has highrecurrence.The occurrence and development and relapse mechanism of bladdercancer is still unknown at present,In recent years,with the molecular levelresearch deeply,research findings suggest that abnormal TGF-β1signalingpathway is closely related to many of the epithelial cells of the tumoroccurrence and development.Objective:To selecte the best inhibiting effect ofTGF-β1siRNA which transfected into the EJ cell lines,studying its effects onbladder tumor angiogenesis.Methods: The EJ cell lines are developed,thelogarithmic phase of EJ cell lines are divided into the conventional cellgroup,the liposome group (negative control) and the three group (group A andgroup B and C) with different TGF-β1siRNA expression vector group.Threekinds of designed TGF-β1siRNA are Chosen,they are respectively transfectioninto group A and group B and group C by plasmid.After transfection cellculture24hours,real-time PCR and Elisa are used to detect TGF-β1expression that in the EJ cell lines which transfected the TGF-β1siRNA,andselecting the best inhibiting effect group.The logarithmic phase of EJ cell linesare transfected and divided into the EJ cells group,the control group (TGF-beta1), the recombinant plasmid group (TGF-beta1sirna expression vectorgroup),The control group is stimulated by10ng/ml TGF-beta1.Aftertransfection cell culture24hours,Western Blot is used to detected the VEGFexpression that in the normal EJ cell group,TGF-β1group,the TGF-β1siRNAgroup,and the recombinant plasmid group.Results:The EJ cell lines arecultivate well， the TGF-β1siRNA transfect successfully.The RT-PCRdetection:Compared with the conventional cell group,The expression of TGF- β1mRNA in the liposome group is no statistical significance(P>0.05).It showsthat the blank plasmid has no noticeable interference on TGF-β1mRNAexpression;Compared with the conventional cell group and the liposomegroup,The expression of TGF-β1mRNA has dropped significantly in the threedifferent TGF-β1SiRNA group(P<0.05).The TGF-β1siRNA group C is themost obvious(P<0.01). It Shows that the design of three kinds of TGF-β1SiRNA have inhibition to the expression of TGF-β1mRNA,the TGF-β1siRNA group C is the most obvious.The Elisa detection:Compared with theconventional cell group,The expression of TGF-β1protein in the liposomegroup is no statistical significance(P>0.05).It shows that the blank plasmid hasno noticeable interference on TGF-β1protein expression, the testing result isthe same with RT-PCR.Compared with the conventional cell group and theliposome group,The expression of TGF-β1protein has dropped significantlyin the three different TGF-β1SiRNA group(P<0.05),the testing result is thesame with RT-PCR.The Elisa also confirm that the designed three kinds ofTGF-β1SiRNA have inhibited the expression of TGF-β1.Western Blot istested the groups of VEGF protein,it has found that:VEGF expresses significantincrease in the EJ cells which transfection TGF-beta1plasmid(P<0.05),Tosuggest that the expression of VEGF and TGF-beta1are positivelycorrelated;Comparing with normal EJ cells group and TGF-β1group,Theexpression of VEGF in EJ cells which transfect the TGF-β1siRNA hassignificantly reduced(P<0.05).Conclusion: The expression of VEGF protein ofthe EJ cells which has transfected TGF-β1siRNA plasmid has significantlyreduced, Thus,That affirms the TGF-β1siRNA have obvious inhibitory effecton bladder tumor angiogenesis.