Song Yue, Kong Bei-hua, Liu Pei-shu, Ma Dao-xin, Qu Xun, Jiang Sen
Department of Obstetrics and Gynocology, Qilu Hospital of Shandong University, Jinan 250012, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Aug;25(4):438-42.
To investigate the in vitro effect of HSV-tk/GCV using a hTERT promoter-driven vector system on Skov3 ovarian cancer cells.
An expression vector (pBTdel-279-tk) containing tk gene under the hTERT promoter was constructed by molecular biological methods, and then was transfected into Skov3 ovarian cancer cells, normal ovarian epithelial cells (NOEC) and human embryonic lung fibroblast by cationic liposome. Following the transfection with tk, GCV was added, and MTT and flow cytometry methods were applied to investigate its antitumor effect. RT-PCR was used to detect the tk gene in ovarian cancer cells and normal cells after the transfection of pcDNA3-tk or pBTdel-279-tk.
pBTdel-279-tk/GCV system induced apoptosis in hTERT-positive ovarian cancer cells, but not in hTERT-negative normal ovarian epithelial cells and fibroblasts. The hTERT promoter system was almost as efficient in inducing cancer cell death as the CMV promoter. tk gene was expressed in Skov3 cells and NOEC after pcDNA3-tk transfection, while positive was only in ovarian cancer cells after pBTdel-279-tk transfection.
The telomerasespecific transfer of the tk gene under the hTERT promoter is a novel targeting approach for the treatment of ovarian cancer and may lead to an effective and specific gene therapy.
利用人端粒酶逆转录酶（hTERT）启动子驱动的载体系统研究单纯疱疹病毒胸苷激酶（HSV-tk）/丙氧鸟苷（GCV）对Skov3卵巢癌细胞的体外作用。
采用分子生物学方法构建hTERT启动子调控下含tk基因的表达载体（pBTdel-279-tk），然后通过阳离子脂质体转染Skov3卵巢癌细胞、正常卵巢上皮细胞（NOEC）和人胚肺成纤维细胞。转染tk后加入GCV，采用MTT法和流式细胞术检测其抗肿瘤作用。转染pcDNA3-tk或pBTdel-279-tk后，用RT-PCR检测卵巢癌细胞和正常细胞中的tk基因。
pBTdel-279-tk/GCV系统可诱导hTERT阳性的卵巢癌细胞凋亡，但对hTERT阴性的正常卵巢上皮细胞和成纤维细胞无此作用。hTERT启动子系统诱导癌细胞死亡的效率与巨细胞病毒（CMV）启动子相近。pcDNA3-tk转染后，tk基因在Skov3细胞和NOEC中表达，而pBTdel-279-tk转染后，仅在卵巢癌细胞中呈阳性表达。
hTERT启动子调控下tk基因的端粒酶特异性转移是一种治疗卵巢癌的新型靶向方法，可能带来有效且特异的基因治疗。
