重组腺病毒载体介导的基因治疗及其与放射治疗联合应用对子宫颈癌HeLa细胞的作用  

Experimental study on adenovirus-mediated transfer of p53 or adenovirus-mediated transfer of p53 combined with radiotherapy for human cervical cancer cell line HeLa cells

作　　者：孔为民[1] 孙建衡[1] 林晨[2] 查园园[2] 路国秋[2] 付明[2] 张雪燕[2] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤医院妇瘤科 [2]中国医学科学院肿瘤研究所分子肿瘤学国家重点实验室,北京100021

出　　处：《中华妇产科杂志》2002年第6期352-355,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的　观察重组腺病毒载体 (Ad)介导的野生型p5 3基因 (Ad p5 3)单独及与放射治疗(放疗 )联合应用 (联用 )对宫颈癌HeLa细胞的体内、外作用效果。方法　将Ad p5 3单独或与放疗联用 ,通过逆转录聚合酶链反应 (RT PCR)技术证实转染的成功性 ;通过细胞生长和存活的抑制实验、流式细胞仪分析、梯状DNA(DNAladder)实验 ,以及裸鼠皮下移植瘤的治疗实验 ,观察Ad p5 3对宫颈癌HeLa细胞的体内、外作用及可能作用机制。结果　RT PCR技术检测结果显示 ,加入Ad p5 3后 ,p5 3mRNA表达明显增加。在体外实验中 ,Ad p5 3对宫颈癌HeLa细胞生长的抑制率为 6 1 8% ,单独放疗的抑制率为 4 4 6 % ,而Ad p5 3与放疗联用的抑制率为 85 5 % ,后者分别与前两者比较 ,差异均有显著性 (P <0 0 5 )。在体内实验中 ,单独Ad p5 3与放疗对HeLa细胞裸鼠皮下移植瘤生长的抑瘤率分别为5 4 8%和 5 8 4 % ,而Ad p5 3与放疗联用的抑瘤率达到 77 4 % ,后者分别与前两者比较 ,差异均有显著性 (P <0 0 5 )。结论　Ad p5 3在体内、外对宫颈癌HeLa细胞生长均有明显的抑制作用 ,而将其与放疗联用则抑制作用更明显。作用机理可能与Ad p5 3诱导细胞凋亡和引起细胞G2Objective To evaluate the therapeutic efficiency of adenovirus mediated transfer of p53(Ad p53) or Ad p53 combined with radiotherapy for human cervical cancer cell line HeLa cells Methods HeLa cells were either transfected with Ad p53 alone or with Ad p53 combining with radiotherapy Reverse transcription polymerase chain reaction(RT PCR), the cell growth, morphological changes, cell cycle, apoptosis and molecular changes were measured using cell counting, microscopy, flow cytometry, DNA ladder, and in vivo therapy experiments to evaluate the therapeutic efficiency of the regimens Results Ad p53 could be transfected into HeLa cells and p53 mRNA could be expressed more in the targeted HeLa cells Ad p53 transfer had strong therapeutic effects to HeLa cells in vitro and in vivo (the inhibition rates were 61 8% and 54 8%, respectively), morever, the combined administration of Ad p53 and radiotherapy had stronger therapeutic effects in vitro and in vivo (the inhibition rate was 85 5% in vitro, and the inhibition rate was 77 4% in vivo) The inhibition rate by radiotherapy alone was 44 6% in vitro and 58 4% in vivo Massive apoptosis of HeLa cells was induced by these regimens Cell cycle analysis demonstrated that all these regimens could arrested HeLa cells in G 2 M Conclusions After introduced into HeLa cells, Ad p53 shows therapeutic efficiency for HeLa cells both in vitro and in vivo Moreover, Ad p53 could enhance therapeutic efficiency for HeLa cells when combined with radiotherapy

关 键 词：重组腺病毒载体 宫颈肿瘤 HELA细胞 基因疗法 放射疗法 AD-P53 

分 类 号：R737.33[医药卫生—肿瘤]