目的 构建含有鞘脂激活蛋白C(Saposin C)的真核表达载体,探讨Saposin C在前列腺癌雄激素依赖性前列腺细胞(LNCaP)中对雄激素受体(AR)及细胞增殖的影响。方法 通过cDNA法构建含有TAT和Saposin C的真核表达载体,并将其转染至前列腺癌LNCaP细胞。通过MTS、RT-PCR、Western blot等实验,分别检测TAT-Saposin C与Saposin C的AR基因表达与其转录激活功能,以及对前列腺癌细胞增殖的影响。结果 在不依赖雄激素的情况下,Saposin C能够上调AR与PSA基因mRNA和蛋白质水平的表达,增加AR的转录激活活性,刺激LNCaP细胞的增殖。结论 Saposin C通过雄激素非依赖性上调AR的表达与转录激活活性,促进LNCaP细胞的增殖。
Objective To construct eukaryotic expression vectors containing Saposin C so as to investigate the effects of Saposin C on androgen receptor (AR) expression and proliferation of LNCaP cells. Methods Recombination vectors pcDNA-TAT-Saposin C and pcDNA-Saposin C were constructed using cDNA methods. After the vectors were transfected into LNCaP cells, the proliferation of LNCaP cells was assessed by MTS assay. Expression of AR in TAT-Saposin C and Saposin C and transcriptional activation were examined by Western blot and RT-PCR. Results In the absence of androgen, Saposin C up-regulated AR and PSA expressions at the protein and mRNA levels, increased transcriptional activation of AR, and stimulated the proliferation of LNCaP cells. Conclusion Saposin C promotes the proliferation of LNCaP cells by up-regulating AR expression and transcriptional activation in an androgen-independent manner.