Ad—TIMP-3转染增加宫颈癌细胞对顺铂的敏感性

Adenovirus-delivered tissue inhibitor of metalloproteinases-3 transfection increases the sensitivity of cervical cancer ceils to cisplatin

目的研究携带TIMP-3基因的重组腺病毒（Ad-TIMP-3）转染宫颈癌细胞后对顺铂（cDDP）敏感性的影响，探讨Ad-TIMP-3基因与cDDP联合在宫颈癌治疗中的作用。方法Ad-TIMP-3体外感染宫颈癌细胞系后，观察细胞凋亡效应、细胞周期及细胞内p53蛋白表达情况。Ad-TIMP-3与cDDP联合应用后，检测细胞克隆形成率；MTT法检测杀伤率及评价协同效应；裸鼠致瘤实验观察瘤体生长曲线。结果Ad-TIMP-3转染可促进细胞凋亡，引起细胞G2／M期阻滞，并诱导宫颈癌细胞内p53蛋白表达。Ad-TIMP-3与cDDP联合应用后，细胞克隆形成率明显减少，杀伤率成倍提高，肿瘤生长显著抑制。结论Ad-TIMP-3可增加宫颈癌对cDDP化疗的敏感性，二者联合应用在宫颈癌治疗中有潜在的应用价值。

Objective To investigate the effects of adenovirus-delivered tissue inhibitor of metalloproteinases-3 （Ad-TIMP-3） on sensitivity of cervical cancer cells to cisplatin and evaluate the potential application of this combined scheme in cervical cancer treatment. Methods Cells of cervical cancer CaSKi cell line were infected with Ad-TIMP-3 in vitro. Apoptotic effect, cell cycle changes and p53 protein expression were detected. After combined treatment of those cells with cisplatin, colony formation test was performed and cytotoxicity was detected by MTT. The growth curve and tumor growth inhibition in vivo were evaluated. Results The expressions of TIMP-3 mRNA and protein were significantly upregulated after transfection. As a result, massive apoptosis was induced and the cells were arrested at G2/M phase. Exogenous overexpression of TIMP-3 increased p53 protein level markedly in spite of the backgrounds of p53 gene in cells. Combined with cisplatin treatment, the cloning efficiency was decreased. A synergism was observed by isobolic method （ D 〈 1 ） in vitro and tumor growth was significantly inhibited in vivo. Conclusion Ad-TIMP-3 is a powerful proapoptotic agent. It increases sensitivity of the cells to cisplatin and the Ad-TIMP-3 gene therapy in combination with cisplatin could be a promising alternative in cervical cancer treatment.