转基因溶瘤性单纯疱疹病毒对人鼻咽癌细胞裸鼠皮下种植瘤的治疗作用

Therapeutic effect of an oncolytic herpes simplex virus vector against human nasopharyngeal carcinoma cell subcutaneous tumors in nude mice

目的观察转基因溶瘤性单纯疱疹病毒载体G47Δ对人鼻咽癌裸鼠皮下种植瘤的治疗效果,探讨G47Δ对人鼻咽癌的治疗作用。方法体外培养人鼻咽癌细胞CNE2,选取Balb-c裸鼠12只,分为病毒组和对照组,每组6只。于裸鼠双侧皮下注射人鼻咽癌细胞CNE2构建鼻咽癌皮下种植瘤模型,右侧为对照侧,左侧为处理侧。待肿瘤直径约5 mm时,病毒组每周2次于处理侧皮下瘤内注射4×104/μl G47Δ,共注射4次,对照侧不做处理;对照组处理侧每周两次注射磷酸盐缓冲液(PBS),共注射4次,对照侧不做处理。接种后第4、7、11、14、17、21、24、28、32天测量肿瘤最大长径(a)值和最大宽径(b)值,并计算肿瘤体积(V=a×b2/2)。病毒组裸鼠于接种后90 d处死裸鼠,对照组皮下肿瘤最长径达18 mm时处死裸鼠,采用方差分析比较2组裸鼠双侧肿瘤的大小,采用KaplanMeier法比较2组的生存时间。结果病毒载体治疗第21天,2组裸鼠皮下肿瘤体积已有明显差异,病毒组接种后28 d,治疗侧肿瘤体积继续缩小,至接种后32 d,该侧肿瘤消退。病毒组处理侧与病毒组对照侧、病毒组处理侧与对照组处理侧的处理主效应、时间主效应、处理效应与时间效应交互作用均有统计学意义,表明病毒组处理侧的治疗效果优于病毒组对照侧及对照组处理侧。病毒治疗组裸鼠的生存时间明显长于对照组裸鼠,差异具有统计学意义(χ2=12.11,P <0.01)。结论瘤内注射溶瘤性单纯疱疹病毒载体G47Δ可有效治疗鼻咽癌皮下种植瘤。

ObjectiveTo assess the efficacy of a herpes simplex virus （HSV） vector, G47Δ, in the treatment of human nasopharyngeal carcinoma cell subcutaneous tumors in nude mice. MethodsHuman nasopharyngeal carcinoma cells （CNE-2） were cultured in vitro. A bilateral subcutaneous nasopharyngeal tumor model was then established in Balb-c nude mice by subcutaneous injection of CNE2 cells. When tumors were approximately 5 mm in maximal diameter, mice were unilaterally （the left side） inoculated with 50 μL of 2×10^6/μL G47Δ or virus buffer twice a week. Tumor size was measured using an external caliper and tumor volume was calculated （V=a×b^2/2）. When the diameter of tumor reached 18 mm, the mice were sacrificed. Tumor size and survival time were compared between the two groups. ResultsIn the bilateral subcutaneous nasopharyngeal tumor model, compared with the non-virus treatment side, the subcutaneous tumors treated with G47Δ grew slowly and finally disappeared （P 〈 0.01）. The survival time of the virus group was significantly longer than that of the control group （χ2=12.11, P=0.005）. ConclusionIntratumoral inoculation of G47Δ could effectively cure nasopharyngeal carcinoma subcutaneous tumors in nude mice, suggesting that G47Δ may be used for clinical therapy of nasopharyngeal carcinoma if further clinical trials can confirm our findings.