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目的:观察P27基因重组腺病毒联合中药片仔癀对人骨肉瘤裸鼠移植瘤体内生长的抑制作用。方法:①用人骨肉瘤细胞株MG-63接种于裸鼠腋部皮下,连续传代3次,肿瘤生长稳定后,再用组织移植法构建人MG-63细胞裸鼠原位骨肉瘤模型;②肿瘤细胞接种1周肿瘤长至0.7cm左右时,选择40只肿瘤体积均一的裸鼠随机分成5组:空白对照组(8只),PBS溶液注射治疗;P27基因治疗组(8只),raav-P27病毒液注射治疗;Paav-MCS空载体组(8只),raav-MCS病毒注射治疗;片仔癀组(8只),片仔癀溶液灌胃治疗;联合治疗组(8只),raav-P27病毒液注射联合片仔癀灌胃治疗。③观测指标:骨肉瘤裸鼠成瘤率、荷瘤裸鼠生活质量及毒副反应、肿瘤块的体积和肿瘤的生长抑制率。结果:①成功构建了裸鼠原位骨肉瘤模型;②单纯灌服片仔癀溶液、P27基因治疗及片仔癀、P27基因联合治疗组肿瘤的生长明显受到抑制,与空白对照组及空载体组差异显著(P<0.05),肿瘤生长抑制率分别达32.24%,55.25%与61.84%。空白对照组与空载体组相比,无显著差异(P>0.05);③荷瘤裸鼠一般情况良好,活动能力较强。结论:①腺相关病毒介导的p27基因能有效抑制人骨肉瘤MG-63的生长。②P72基因与片仔癀联合应用后,产生协同作用,其抑瘤效果与改善荷瘤裸鼠的生存情况比单用片仔癀治疗或单用P27基因治疗更明显。③中药片仔癀对提高荷瘤裸鼠的生活质量有一定的帮助。
Objective: To observe the inhibitory effect of P27 recombinant adenovirus combined with Pien Tze Huang on the growth of human osteosarcoma xenografts in nude mice. Methods: ① Human osteosarcoma cell line MG-63 was subcutaneously inoculated subcutaneously in the axilla of nude mice for three times in succession. After the tumor grew stably, the in vivo osteosarcoma model of human MG-63 cells was constructed by tissue transplantation. One week after inoculation, the nude mice with a uniform volume of tumor were randomly divided into 5 groups: control group (n = 8), PBS solution injection treatment; P27 gene therapy group (n = 8), raav- P27 virus liquid injection therapy; Paav-MCS empty vector group (8), raav-MCS virus injection treatment; Pien Tze Huang group (8), Pien Tze Huang solution gavage treatment; combination therapy group (8), raav-P27 virus solution Injection joint Pien Tze Huang irrigation treatment. ③Observation: The rate of tumor formation in nude mice with osteosarcoma, the quality of life and toxicity of tumor-bearing nude mice, the volume of tumor mass and tumor growth inhibition rate. Results: ① The model of orthotopic osteosarcoma was successfully established in nude mice; ② The growth of tumor was inhibited obviously by P27 gene therapy and P27 gene therapy combined with P27 gene therapy group, which was significantly different from that of blank control group and empty vector group P <0.05). The tumor growth inhibition rates were 32.24%, 55.25% and 61.84% respectively. There was no significant difference between blank control group and empty vector group (P> 0.05). (3) The tumor-bearing nude mice were generally in good condition with strong activity ability. Conclusion: (1) Adeno-associated virus-mediated p27 gene can effectively inhibit the growth of human osteosarcoma MG-63. ②P72 gene and Pien Tze Huang joint application, have a synergistic effect, and its anti-tumor effect and improve the survival of tumor-bearing nude mice than Pien Tze Huang treatment alone or P27 gene therapy alone is more obvious. ③ Pien Tze Huang Chinese medicine to improve the quality of life of nude mice have some help.