Showing papers in "Journal of Contemporary Urologic and Reproductive Oncology in 2011"

Construction of dsRNA expression vector and research of RNA activation

Abstract: Objective To construct the small-molecule double-stranded RNA (dsRNA) expression vector plasmid,and to investigate its activation ability of the expression of cell cycle repressor protein p21WAF1/CIP1(p21) in human prostate cancer cell line PC-3 in vitro.Methods Synthetic the sequence-specific corresponding DNA fragments of the promoter with the p21 gene,as well as the same length of random DNA fragments,by means of the recombinant DNA technology to construct these fragments into the eukaryotic expression plasmids pGenesil-1,constructed with the purpose of the expression of dsRNA fragments plasmid vectors (dsRNAp21-pGenesil-1) and the random fragments of the control dsRNA expression vectors (dsRNACon-pGenesil-1).The dsRNAp21-pGenesil-1 (experimental group),dsRNACon-pGenesil-1 (negative control group) and a blank plasmid pGenesil-1 (control group) were transfected into human prostate cancer cell line PC-3 in vitro respectively with lipofectamine reagent,and transfection efficency was detected by laser scanning confocal microscope.Immunohistochemistry and reverse transcription-PCR were performed to examine the difference expression level of p21 in each group.Results These dsRNAp21-pGenesil-1 and the dsRNACon-pGenesil-1 eukaryotic cell expression vectors were successfully constructed.For each of the groups of plasmids transfected into PC-3 cells,p21 gene expression in the experimental group significantly increased,while there was no significant difference between the negative control group and the blank control group.Conclusions The dsRNAp21-pGenesil-1 express dsRNA molecule successfully,and active the level of expression of p21 mRNA and protein,and provide an effective tool for studying the activation effect of dsRNA.

Peptides CSNRDARRC affinity and bingding sites study of bladder cancer markers

Abstract: Objective To research the affinity of peptides CSNRDARRC with human T24 bladder cancer cell.MethodsThe binding ability of peptides were determined by FACS and the binding sites were determined by confocalmicroscope.ResultsFlowcytometry showed that peptides CSNRDARRC bind to T24 in a concentration-dependent manner,the laser scanning confocal fluorescence microimaging system confirmed that peptides CSNRDARRC was located in Nucleus.ConclusionsPeptides CSNRDARRC have high affinity with human T24 bladder cancer cell and have the binding sites in Nucleus.This experiment improved some experimental basis and foundation for using the peptides in vivo characterization.

Expression and significance of E-cadherin and Bcl-2 in early renal carcinoma and adjacent tissues of different levels of distance

Abstract: Objective To test the expression of E-candherin and Bcl-2 in early renal carcinoma and adjacent tissues,provide a theoretical basis for the safety margins in nephron sparing surgery.MethodsThe clinical data of 45 patients who were undertaken radical nephrectomy in the First Affiliated Hospital of Henan Scientific and Technological University,pathological diagnosis were all renal clear cell carcinoma.Taken cancer tissue(a)、the different distance of adjacent tissues 0.5 cm(b)、1.0 cm(c)、2.0 cm(d),use the type of non-biotin immunohistochemical Elivision method to study the different expression about E-candherin and Bcl-2 in different organizations.ResultsThe expression of E-candherin in renal carcinoma was 11.1%,Bcl-2 was 80.0%.The positive rate of expression about E-candherin and Bcl-2 were statistically significantly different between group a and group b、c and d(P0.01),but the difference between group b、c and d were not statistically significant(P0.05).ConclusionsThe safety margin in nephron sparing surgery for the early renal carcinoma is at least 0.5 cm of normal parenchyma tissue beyond the pseudocapsule of the tumor.