中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (4): 251-254.

• 论著 • 上一篇    下一篇

携带IL-18基因的溶瘤腺病毒对裸鼠恶性黑素瘤移植瘤的抗肿瘤作用

刘彦群1,杨春华1,毛立军2,裴东生2,魏志平1,郑骏年2   

  1. 1. 徐州医学院附属医院皮肤科
    2. 徐州医学院肿瘤重点实验室
  • 收稿日期:2008-05-19 修回日期:2008-06-16 出版日期:2009-04-15 发布日期:2009-04-14
  • 通讯作者: 郑骏年，zhengjunnian@sina.com
  • 基金资助:

    国家自然科学基金（30570385，30700999）;国家教委归国人员科研启动基金资助项目（编号）

Antitumor effects of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice

  • Received:2008-05-19 Revised:2008-06-16 Online:2009-04-15 Published:2009-04-14

PDF (PC)

328

赞

摘要：

目的 探讨携带IL-18基因的溶瘤腺病毒（ZD55-IL-18）对裸鼠恶性黑素瘤移植瘤的抗肿瘤效果。方法 取A375细胞2 × 106个接种于BALB/c裸鼠腋下组织内，建立裸鼠黑素瘤模型，当移植瘤体积达100 ~ 150 mm3时将其随机分为3组，每组8只，分别于瘤体内注射ZD55-IL-18、Ad-IL-18和磷酸盐缓冲液（PBS）。通过定期测量肿瘤体积观察肿瘤生长抑制情况，并切取肿瘤进行HE染色，观察肿瘤细胞生长形态；激光共聚焦显微镜下观察移植瘤组织IL-18表达水平、E1A蛋白的表达情况。免疫组化检测荷瘤裸鼠肿瘤组织CD34染色标记测定的肿瘤组织微血管密度；TUNEL法检测移植瘤组织细胞凋亡情况。结果 肿瘤生长曲线表明，ZD55-IL-18处理组肿瘤生长速度较PBS组明显减慢，接种44 d后ZD55-IL-18组肿瘤平均体积为（1039.378 ± 29.67） mm3，Ad-IL-18组为（2900.46 ± 62.65） mm3，PBS组为（3980.24 ± 63.78） mm3，各组间差异有统计学意义（P < 0.01）。HE染色发现，ZD55-IL-18处理组细胞核深染，很少见核仁，显示肿瘤组织生长抑制。激光共聚焦结果表明，ZD55-IL-18处理组IL-18的表达阳性率比Ad-IL-18处理组明显增强（P < 0.01），并可以在肿瘤组织中表达E1A蛋白；免疫组化检测显示，ZD55-IL-18治疗组肿瘤微血管密度明显低于PBS组（P < 0.05）。不同处理组移植瘤细胞凋亡阳性率ZD55-IL-18处理组（86.28% ± 3.25%） ＞ Ad-IL-18处理组（43.67% ± 3.46%） ＞ PBS处理组（10.73% ± 2.48%），各组之间差异有统计学意义（P ＜ 0.05）。结论 携带IL-18基因的溶瘤腺病毒ZD55-IL-18对裸鼠恶性黑素瘤移植瘤的生长及转移有抑制作用，并探讨了其可能的作用环节。

关键词: 溶瘤腺病毒；IL-18；恶性黑色素瘤

Abstract:

Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice. Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma. When the volume of implanted tumor reached 100 - 150 mm3, murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18, IL-18 gene-expressing adenovirus named Ad-IL-18, phosphate buffer saline （PBS）, respectively. The tumor size was measured at an interval of 4 days for 9 weeks. Hematoxylin and eosin （HE） staining was performed to observe the morphological changes of tumor cells. The protein expression of IL-18 and E1A，microvessel density in tumor tissue，and apoptosis of tumor xenografts were detected by immuno- fluorescence assay, immunohistochemistry and in situ end labeling technique （TUNEL）, respectively. Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor. Forty-four days after the treatment, the mean tumor volume was 1039.378 ± 29.67 mm3 in ZD55-IL-18-treated mice, significantly smaller than that in Ad-IL-18 treated mice （2900.46 ± 62.65 mm3） and PBS-treated mice （3980.24 ± 63.78 mm3）. HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice. Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs. AD55-IL-18-treated mice （83.4% ± 3.2% vs 24.4% ± 2.1%, P < 0.01）. Moreover, immunofluorescence assay revealed the presence of E1A protein in tumor tissue. A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice （P < 0.01）. The apoptosis rate in tumor cells from high to low was 86.28% ± 3.25% in ZD55-IL-18-treated mice, 43.67% ± 3.46% in Ad-IL-18-treated mice, and 10.73% ± 2.48% in PBS-treated mice; there was a significant difference between the three groups （all P < 0.05）. Conclusion The oncolytic adenovirus expressing human IL-18 gene, ZD55-IL-18, has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.