摘要
Objective To explore the growth inhibiting effects on human bladder cancer by antisense RNA targeting the proliferating cell nuclear antigen (PCNA) gene. Methods The eukaryotic expression vector for antisense PCNA cDNA was constructed and transferred into a bladder cancer EJ cell line. The PCNA expression in the cancer cells was detected by RT-PCR and Western blotting assays. The in vitro proliferation activities of the transferred cells were observed by growth curve,tetrazolium bromide (MTT) colorimetry,tritiated thymidine ( 3H-TdR)incorporation, flow cytometry and clone formation testing,while its in vivo anti-tumor effects were detected on nude mice allograft models.Results After the antisense vector,pLAPSN,was transferred,cellular PCNA expression was inhibited at both protein and mRNA levels. The growth rates of EJ cells were reduced from 27.91% to 62.07% ( P <0.01),with an inhibition of DNA synthesis rate by 52.31% ( P <0.01). Transferred cells were blocked at G 0/G 1 phases in cell-cycle assay,with the clone formation ability decreased by 50.81% ( P <0.01). The in vivo carcinogenic abilities of the transferred cancer cells were decreased by 54.23% ( P <0.05). Conclusions Antisense PCNA gene transfer could inhibit the growth of bladder cancer cells in vitro and in vivo,which provided an ideal strategy for gene therapy of human cancers.
Objective To explore the growth inhibiting effects on human bladder cancer by antisense RNA targeting the proliferating cell nuclear antigen (PCNA) gene. Methods The eukaryotic expression vector for antisense PCNA cDNA was constructed and transferred into a bladder cancer EJ cell line. The PCNA expression in the cancer cells was detected by RT-PCR and Western blotting assays. The in vitro proliferation activities of the transferred cells were observed by growth curve,tetrazolium bromide (MTT) colorimetry,tritiated thymidine ( 3H-TdR)incorporation, flow cytometry and clone formation testing,while its in vivo anti-tumor effects were detected on nude mice allograft models.Results After the antisense vector,pLAPSN,was transferred,cellular PCNA expression was inhibited at both protein and mRNA levels. The growth rates of EJ cells were reduced from 27.91% to 62.07% ( P <0.01),with an inhibition of DNA synthesis rate by 52.31% ( P <0.01). Transferred cells were blocked at G 0/G 1 phases in cell-cycle assay,with the clone formation ability decreased by 50.81% ( P <0.01). The in vivo carcinogenic abilities of the transferred cancer cells were decreased by 54.23% ( P <0.05). Conclusions Antisense PCNA gene transfer could inhibit the growth of bladder cancer cells in vitro and in vivo,which provided an ideal strategy for gene therapy of human cancers.
基金
ThisstudywassupportedbyNationalNatureScienceFoundationofChina (No 3 9770 73 9)
