中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (2): 91-94.

• 论著 • 上一篇    下一篇

携带IL-24基因的溶瘤腺病毒诱导黑素瘤细胞凋亡

蒋冠1,刘彦群1,郑骏年2,魏志平1,毛立军2,裴冬生2   

  1. 1. 徐州医学院附属医院皮肤科
    2. 徐州医学院肿瘤生物治疗重点实验室
  • 收稿日期:2008-09-23 修回日期:2009-05-18 出版日期:2010-02-15 发布日期:2012-03-30
  • 通讯作者: 蒋冠 E-mail:5721268@163.com
  • 基金资助:
    30700999;30570385;国家自然科学基金资助项目（编号）

Oncolytic adenoviruses harboring IL-24 gene induce the apoptosis of melanoma cells

  • Received:2008-09-23 Revised:2009-05-18 Online:2010-02-15 Published:2012-03-30

PDF

351

赞

摘要： 目的 探讨携带IL-24基因的溶瘤腺病毒（ZD55-IL-24）诱导人黑素瘤A375细胞凋亡的作用。方法 用PCR方法鉴定ZD55-IL-24，并大量扩增、纯化和病毒滴度测定。将ZD55-IL-24感染人黑素瘤A375细胞。结晶紫染色观察细胞毒性，MTT法测定细胞存活率，Western印迹检测溶瘤腺病毒E1A、IL-24基因蛋白表达。结果 PCR鉴定表明ZD55-IL-24包含IL-24基因且没有野生型腺病毒污染；结晶紫染色结果表明ZD55-IL-24对A375细胞有明显的细胞毒性效应。MTT法结果显示，ZD55-IL-24对A375细胞的增殖抑制作用呈时间和浓度依赖性效应。Western印迹结果表明ZD55-IL-24能在A375细胞内高效表达E1A、IL-24基因。结论 溶瘤腺病毒ZD55-IL-24能携带IL-24基因在黑素瘤A375细胞中高效表达，抑制细胞增殖并诱导其凋亡。

关键词: 黑色素瘤, 基因,IL-24, 溶瘤病毒

Abstract: Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene （ZD55-IL-24） on the apoptosis of human melanoma cell line A375. Methods The oncolytic adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-IL-24, ZD55 fused with enhanced green fluorescent protein （ZD55-EGFP）, and replication-deficient adenovirus ZD55 carrying IL-24 gene （AD-IL-24）， respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively. Subsequently, the cytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium （MTT） assay, respectively. The expressions of E1A and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirus contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious cytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time- and concentration-dependent manner. As shown by Western blot analysis, ZD55-IL-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Key words: Genes,IL-24