IL-12基因联合CD40L基因治疗小鼠黑色素瘤的实验研究

Studies on the effects of Combined IL-12 and CD40L gene therapy for murine melanoma

目的:观察腺病毒介导的小鼠白细胞介素12基因(AdmIL-12)和CD40配体基因(AdmCD40L)对小鼠黑色素瘤的抗肿瘤效果。方法:利用B16细胞皮下注射C57BL/6小鼠建立小鼠黑色素瘤模型,单独或联合应用分别携带小鼠IL-12基因和CD40L基因的重组腺病毒进行直接瘤内注射治疗,观察小鼠皮下肿瘤生长及成活情况。采用乳酸脱氢酶释放法检测荷瘤小鼠脾细胞CTL活性变化。结果:AdmIL-12和AdmCD40L在体内外均能有效表达;AdmIL-12能显著抑制荷瘤小鼠皮下肿瘤的生长并明显延长其生存时间,能显著增强荷瘤小鼠的脾细胞CTL杀伤活性。AdmCD40L的抗瘤效果不明显,但与Ad-mIL-12联合应用可明显提高抗肿瘤效果。结论:腺病毒介导的mIL-12基因对小鼠黑色素瘤有显著的治疗效果,且与CD40L基因联合应用能进一步提高抗肿瘤效果。

Objective：To observe the therapeutic efficiency of adenovirus-mediated interleukin-12 gene（AdmIL-12） and CD40L ligand gene（AdmCD40L） intratumoral transfer in established murine melanoma in vivo. nethods：C57BL/6 mice were inoculated subcutaneously with B16 cells to establish the murine melanoma model. The tumor-bearing mice were injected intratumorally with murine IL-12 gene and CIMOL gene recombinant adenovirus. Tumor growth and the survival of tumor-bearing mice were observed. The CTL activity was measured in vitro by lactate dehydrogenase（LDH） release assay. Results：Both AdmIL-12 and AdmCD40L can be efficiently expressed in vitro and in vivo. The treatment with AdmIL-12 could significantly inhibit the tumor growth and prolong the survival period of the tumor-beraing mice. Splenic CTL activity of the mice was also enhanced after IL-12 gene transfer. But the anti-tumor effects of AdmCD40L gene were not significant. In contrast, Co-delivery of IL-12 gene and CD40L gene lead to stronger antitumor effects than IL-12 gene alone. Conclusion：Adenovirus-mediated interleukin-12 gene and CD40L ligand gene transfer together intratumorally has significant therapeutic effects on mice melanoma in vivo.