南方医科大学学报

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (12): 1367-1372.

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抗HPV16E6核酶对宫颈癌CaSKi细胞增殖和侵袭力的影响

刘柯1, 王莉丽2, 孙芳1, 郑燕芳3, 张积仁3   

  1. 1. 广州市花都区人民医院妇产科, 广东, 广州, 510800;
    2. 南方医科大学珠江医院妇产科, 广东, 广州, 510282;
    3. 南方医科大学珠江医院肿瘤中心, 广东, 广州, 510282
  • 出版日期:2004-12-20 发布日期:2004-12-20
  • 基金资助:
    收稿日期:2004-5-23。
    基金项目:广东省自然科学基金(96058);
    作者简介:刘柯(1976- ),主治医师,医学硕士,主要从事子宫颈癌分子生物学治疗的研究,电话:020-80910276,E-mail:liuke76@yahoo.com.cn

Effect of anti-HPV16 E6-ribozyme on cell proliferation and invasiveness of cervical carcinoma cell line CaSKi

LIU Ke1, WANG Li-li2, SUN Fang1, ZHENG Yan-fang3, ZHANG Ji-ren3   

  1. 1. 广州市花都区人民医院妇产科, 广东, 广州, 510800;
    2. 南方医科大学珠江医院妇产科, 广东, 广州, 510282;
    3. 南方医科大学珠江医院肿瘤中心, 广东, 广州, 510282
  • Online:2004-12-20 Published:2004-12-20

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摘要： 目的 研究特异性抗HPV16E6核酶对宫颈癌CaSKi细胞增殖和侵袭能力的影响。方法 以脂质体法将抗HPV16E6核酶、空载体质粒分别导入CaSKi细胞,分别为CaSKi-R和CaSKi-P。观察细胞的生长状态,测定CaSKi,CaSKi-R和CaSKi-P3种细胞的生长曲线、软琼脂克隆形成率、细胞侵袭力实验、裸鼠体内致瘤性,以及RT-PCR检测细胞中COX-2、血管内皮生长因子(VEGF)的表达,同时,免疫组化检测3种细胞COX-2、VEGF抗原表达,分析特异性抗HPV16E6核酶对宫颈癌CaSKi细胞增殖和侵袭力的影响。结果 CaSKi、CaSKi-P细胞生长速率、软琼脂克隆形成率、致瘤性和侵袭力相近,而CaSKi-R细胞的细胞生长速率、软琼脂克隆形成率、致瘤性和侵袭力与以上两种细胞相比却明显降低。RT-PCR检测CaSKi-R细胞的COX-2、VEGF表达水平也低于CaSKi、CaSKi-P细胞的表达。细胞免疫组化测定CaSKi、CaSKi-P和CaSKi-R细胞都有COX-2、VEGF抗原表达,而CaSKi-R细胞的抗原染色程度较弱。结论 特异性抗HPV16E6核酶的导入降低了宫颈癌CaSKi细胞的增殖和侵袭表型,可能与宫颈癌CaSKi细胞内COX-2、VEGF的表达水平降低有关。

Abstract: Objective To investigate the effects of anti-HPV16-ribozyme on the cell proliferation and invasiveness of cultured cervical cancer cell line CaSKi. Methods CaSKi cells were transfected with anti-HPV16 E6-ribozyme and empty eukaryotic expression plasmids via lipofectin and designated as CaSKi-R and CaSKi-P cells respectively. The growth rate, cell colony-forming ability on soft agar, invasiveness and tumorigenicity of CaSKi-R, CaSKi-P, and CaSKi cells were studied using corresponding methods. The expressions of cox-2 and vascular endothelial growth factor (VEGF) mRNA of the 3 cell strains were determined with one-step reverse transcriptional PCR (RT-PCR), and immunocytochemistry was employed for detecting the expressions of COX-2 and VEGF antigens. Results No distinct differences in the growth rate, colony-forming ability on soft agar, cell invasiveness and tumorigenicity were observed between CaSKi and CaSKi-P cells, whereas by comparison, CaSKi-R cells exhibited decreased growth rate, colony-forming ability, the cell invasiveness and tumorgenicity, with also lowered expression levels of cox-2 and VEGF mRNA as shown by RT-PCR analysis. Expressions of COX-2 and VEGF antigens were detected in all the 3 cell strains immunocytochemically, but compared with CaSKi and CaSKi-P cells, the antigen expressions in CaSKi-R cells were significantly weaker. Conclusion Anti-HPV16 E6-ribozyme may partially inhibit the prolifera- tion and reduce the invasiveness of CaSKi cells possibly through decreasing cox-2 and VEGF expressions, which are the important agents for tumor invasion.

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