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目的:探讨携带野生型PTEN基因的重组腺病毒(Ad-PTEN)进行子宫内膜癌基因治疗的可行性。方法:BALB/c裸鼠随机分3组,每组8只,以空载体Ad-CMV转染或未转染作为对照组,观察Ad-PTEN体外转染后体内接种对子宫内膜癌细胞致瘤能力的影响。应用LacZ作为报告基因,X-gal染色检测Ad-PTEN体内转染效率。皮下荷人子宫内膜癌BALB/c裸鼠15只,随机分为对照组、Ad-CMV组及Ad-PTEN治疗组,每组5只,每只裸鼠皮下有2个移植瘤。待皮下移植瘤长至4~5 mm时,治疗组Ad-PTEN肿瘤内注射,每个移植瘤5×108pfu/100μl,隔日1次,共3次,观察裸鼠肿瘤体积的变化及有无不良反应,最后1次治疗后15 d处死裸鼠,取肿瘤组织及治疗组裸鼠肝脏病理检查。结果:Ad-PTEN体外转染子宫内膜癌RL 95-2细胞,子宫内膜癌细胞致瘤能力完全抑制,裸鼠成瘤率为0;而空载体转染组和PBS对照组裸鼠成瘤率均为100%。重组腺病毒在裸鼠体内具有较高的转染效率,Ad-CMV-LacZ肿瘤内注射后96 h,转染效率可达到80%。Ad-PTEN肿瘤内注射可使裸鼠皮下移植瘤生长速度显著减慢,肿瘤体积较对照组明显缩小(P<0.05),Ad-PTEN基因治疗未发现明显不良反应。结论:PTEN基因治疗有可能成为子宫内膜癌非手术治疗的一个新方法,具有潜在的应用价值。
Objective: To investigate the feasibility of endometrial cancer gene therapy using recombinant adenovirus carrying wild-type PTEN gene (Ad-PTEN). Methods: BALB / c nude mice were randomly divided into 3 groups, 8 in each group. The Ad-PTEN transfected cells were transfected with Ad-PTEN Tumor capacity effects. LacZ was used as a reporter gene and the transfection efficiency of Ad-PTEN was detected by X-gal staining. Fifteen BALB / c nude mice bearing subcutaneously human endometrial carcinoma were randomly divided into control group, Ad-CMV group and Ad-PTEN treatment group, with 5 mice in each group. There were 2 xenografts in nude mice. When the subcutaneously transplanted tumor grows to 4 ~ 5 mm in length, the treatment group is injected with tumor of Ad-PTEN, each transplanted tumor is 5 × 108pfu / 100μl, every other day for 3 times to observe the change of the tumor volume of the nude mice and whether there is any defect The nude mice were killed 15 days after the last treatment, and the pathological examination of the liver in nude mice was performed. Results: Ad-PTEN transfected endometrial carcinoma RL 95-2 cells in vitro, tumorigenic ability of endometrial cancer cells was completely inhibited, the nude mice tumorigenicity rate was 0; while the empty vector transfected group and PBS control group nude mice Tumor rates were 100%. The recombinant adenovirus had higher transfection efficiency in nude mice, and the transfection efficiency could reach 80% after 96 h in Ad-CMV-LacZ tumor. Ad-PTEN intratumoral injection of nude mice subcutaneously tumor growth significantly slowed down, the tumor volume was significantly reduced compared with the control group (P <0.05), Ad-PTEN gene therapy found no significant adverse reactions. Conclusion: PTEN gene therapy may become a new method of non-surgical treatment of endometrial cancer with potential value.