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目的探讨携带基质金属蛋白酶组织抑制因子-3(TIMP-3)基因的重组腺病毒(Ad-TIMP-3)在体外对人乳头瘤病毒(HPV)阳性宫颈癌细胞的作用以及与放疗联合应用的效果。方法 2008年10月至2009年4月采用RT-PCR法和Western Blot法检测感染Ad-TIMP-3的HeLa-Luc与CaSki细胞表达TIMP-3 mRNA及蛋白质水平;MTT法检测Ad-TIMP-3对HeLa-Luc和CaSki细胞增殖的抑制作用;平板克隆形成、细胞黏附性及侵袭性实验检测受X射线辐射细胞生长、黏附性和侵袭的能力。结果感染Ad-TIMP-3后,HeLa-Luc和CaSki细胞表达TIMP-3水平明显升高。Ad-TIMP-3能够明显抑制HeLa-Luc和CaSki细胞增殖,且具有剂量效应关系。Ad-TIMP-3与X射线联合应用可显著减少HeLa-Luc和CaSki细胞平板克隆形成数(P<0.01),显著减弱宫颈癌细胞的侵袭能力及黏附性(P<0.001)。结论 Ad-TIMP-3具有抑制宫颈癌细胞增殖的能力;与X射线联合应用后,Ad-TIMP-3显著提高宫颈癌细胞对放射治疗的敏感性。
Objective To investigate the effect of Ad-TIMP-3 carrying TIMP-3 gene on human papillomavirus (HPV) -positive cervical cancer cells in vitro and its application in combination with radiotherapy effect. Methods The expression of TIMP-3 mRNA and protein in HeLa-Luc and CaSki cells infected with Ad-TIMP-3 was detected by RT-PCR and Western Blot method from October 2008 to April 2009. The expressions of Ad-TIMP-3 On HeLa-Luc and CaSki cell proliferation inhibition; plate clone formation, cell adhesion and invasiveness experiments were detected by X-ray radiation cell growth, adhesion and invasion ability. Results The expression of TIMP-3 in HeLa-Luc and CaSki cells was significantly increased after Ad-TIMP-3 infection. Ad-TIMP-3 can significantly inhibit the proliferation of HeLa-Luc and CaSki cells with dose-effect relationship. The combined application of Ad-TIMP-3 and X-rays significantly reduced the number of platelet colonies (P <0.01) and significantly decreased invasiveness and adhesion of cervical cancer cells (P <0.001). Conclusions Ad-TIMP-3 can inhibit the proliferation of cervical cancer cells. Combined with X-ray, Ad-TIMP-3 can significantly increase the sensitivity of cervical cancer cells to radiation therapy.