Effects of sense tankyrase transfection on smooth muscle cells of corpus cavernosum in rat.
This study was conducted to construct the eukaryotic expression vectors for sense tankyrase and to investigate the effects of tankyrase transfection on smooth muscle cells of corpus cavernosum in the rat. After the eukaryotic expression vectors for sense tankyrase were constructed and identified smooth muscle cells of rat corpus cavernosum were transfected with the recombinant plasmids of sense tankyrase (pcDNA-TNKS). Levels of DNA and RNA were then evaluated. Measurement of telomerase activity was conducted by TRAP-ELISA assay, the length of telomere by Southern blot and the growth curve by MTT assay. We have found that the eukaryotic expression vectors for sense tankyrase were constructed and the recombinant plasmids of sense tankyrase were transfected into smooth muscle cells of rat corpus cavernosum successfully; no significant differences in telomerase activity were observed between TNKS-transfected cells (SMC-TANKS), zero-load- transfected cells (SMC-Zeo), and non-transfected cells (SMC) (P > 0.05). The length of telomere in SMC-TANKS was longer than that in SMC-Zeo or SMC (P < 0.01), and the OD value of TNKS-transfected cell was significantly higher than that of the non-transfected cells (P < 0.01). These results suggested that the eukaryotic expression vectors for sense tankyrase were constructed successfully, which provides the basis for gene therapy. Transfection of recombinant plasmids of sense tankyrase helps change the telomerase length of smooth muscle cells of corpus cavernosum and extend the cell life span.
Wu
X-J
XJ
Urinary Surgery Research Center, Southwest Hospital, Third Military Medical University, Chongqing, PR China. swhxj_wu@sina.com
Song
B
B
Jin
X-Y
XY
Zhang
J-H
JH
D000818:Animals; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D004274:DNA, Recombinant / Q000032:analysis; D007172:Erectile Dysfunction / Q000201:enzymology / Q000473:pathology*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000187:drug effects*; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats / Q000235:genetics; D011993:Recombinant Fusion Proteins; D038501:Tankyrases / Q000494:pharmacology*; D019098:Telomerase / Q000378:metabolism; D016615:Telomere; D014162:Transfection
D004274:DNA, Recombinant; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D038501:Tankyrases; D019098:Telomerase