EFFECTS OF SENSE TANKYRASE TRANSFECTION ON SMOOTH MUSCLE CELLS OF CORPUS CAVERNOSUM IN RAT
EFFECTS OF SENSE TANKYRASE TRANSFECTION ON SMOOTH MUSCLE CELLS OF CORPUS CAVERNOSUM IN RAT
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DOI：
10.1080/01485010500315727
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发表时间：
2006-01
期刊：
影响因子：
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通讯作者：
X-J Wu;B. Song;X-Y Jin;J.H. Zhang
中科院分区：
文献类型：
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作者：
X-J Wu;B. Song;X-Y Jin;J.H. Zhang
本研究构建正义Tankyrase真核表达载体，并观察Tankyrase基因对大鼠海绵体平滑肌细胞的影响。构建正义tankyrase真核表达载体并进行鉴定后，将正义tankyrase真核表达载体(pcDNA-TNKS)导入大鼠海绵体平滑肌细胞。然后对DNA和RNA水平进行评估。TRAP-EL ISA法检测端粒酶活性，Southern印迹法检测端粒长度，四甲基偶氮唑盐(四甲基偶氮唑盐)法测定细胞生长曲线。我们发现正义Tankyrase真核表达载体构建成功，并成功地将正义Tankyrase重组载体导入大鼠海绵体平滑肌细胞，TnKS转染组(SMC-TANKS)、空负荷转染组(SMC-Zeo)和未转染组(SMC)端粒酶活性差异无统计学意义(P>；0.05)。SMC-TANK细胞的端粒长度明显长于SMC-Zeo或SMC细胞(P<；0.01)，且TnKS转染组细胞的OD值显著高于未转染组(P<；0.01)。这些结果表明，正义Tankyrase真核表达载体构建成功，为基因治疗奠定了基础。转导正义Tankyrase重组质粒有助于改变海绵体平滑肌细胞的端粒酶长度，延长细胞寿命。
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.