Zhang Ying, Xiang Yang, Lang Jing-He, Qian Hai-Li, Lin Chen
Department of Obstetrics and Gynecology, Hospital Affiliated to Capital University of Medical Sciences, Beijing 100006, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2007 Apr;29(2):246-51.
To explore the effects of adenovirus delivered tissue inhibitor of metalloproteinases-3 (Ad-TIMP-3) on the biological behaviors of cervical cancer cell lines and to evaluate its potential application in cervical cancer gene therapy.
We transferred Ad-TIMP-3 into cervical cancer cells. The TIMP-3 mRNA expression was assessed by RT-PCR, and the TIMP-3 and p53 protein expressions were assessed with Western blot. The apoptotic changes of cells were illustrated with morphology and DAPI staining. The viability of cells was determined with MTT assay. The abilities of in vitro invasion and adhesion were evaluated by the invasion and adhesion assays respectively.
After infection, the TIMP-3 mRNA and protein were significantly upregulated in a time-dependent manner. Overexpression of TIMP-3 markedly increased p53 protein level in spite of the backgrounds of p53 gene in cells. Ad-TIMP-3 infection induced massive apoptosis of cervical cancer cells with a marked bystander effect. The abilities of in vitro invasion and adhesion were inhibited significantly (P < 0.01). The cytotoxicity of Ad-TIMP-3 was significantly stronger than that of Ad-p53 (P < 0.05, P < 0.01).
Ad-TIMP-3 infection has cytotoxic effects on cervical cancer cells and can inhibit the expressions of these malignant phenotypes. Ad-TIMP-3 may be a potentially useful agent for cervical cancer gene therapy.
探讨腺病毒介导的金属蛋白酶组织抑制剂-3（Ad-TIMP-3）对宫颈癌细胞系生物学行为的影响，并评估其在宫颈癌基因治疗中的潜在应用价值。
将Ad-TIMP-3转染至宫颈癌细胞中。采用逆转录聚合酶链反应（RT-PCR）检测TIMP-3 mRNA表达，采用蛋白质免疫印迹法检测TIMP-3和p53蛋白表达。通过形态学观察和4′,6-二脒基-2-苯基吲哚（DAPI）染色显示细胞凋亡变化。采用噻唑蓝（MTT）比色法检测细胞活力。分别通过侵袭实验和黏附实验评估细胞的体外侵袭和黏附能力。
感染后，TIMP-3 mRNA和蛋白呈时间依赖性显著上调。尽管细胞中p53基因背景不同，但TIMP-3的过表达显著提高了p53蛋白水平。Ad-TIMP-3感染诱导宫颈癌细胞大量凋亡，并具有明显的旁观者效应。细胞的体外侵袭和黏附能力均受到显著抑制（P < 0.01）。Ad-TIMP-3的细胞毒性显著强于Ad-p53（P < 0.05，P < 0.01）。
Ad-TIMP-3感染对宫颈癌细胞具有细胞毒性作用，并能抑制这些恶性表型的表达。Ad-TIMP-3可能是一种潜在的宫颈癌基因治疗药物。
