原位基因治疗联合前药热化疗治疗结肠癌肝转移的实验研究

Anti-tumor effect of cytosine deaminase genetherapy in situ combined with thermochemotherapy for colon cancer liver metastasis in nude mice

目的:探讨胞嘧啶脱氨酶(CD)基因联合5-氟胞嘧啶(5-FC)热化疗对裸鼠结肠癌肝转移的治疗效果。方法:30只BALB/c裸鼠经门静脉注射人结肠癌Lovo细胞建立结肠癌肝转移模型,随机分为转基因组和非转基因组,每组15只。转基因组:经腹腔注射病毒上清(0.2mL/d,共5d)进行原位基因转染;非转基因组经腹腔注射生理盐水(0.2mL/d,共5d),2组同时腹腔注射42℃500mg·kg-1.d-1)5-FC。治疗21d后将动物处死,观察肝脏转移率和转移结节数,PCR法检测目的基因在肿瘤组织的整合,RT-PCR法检测目的基因在肿瘤组织的表达,观察肿瘤病理学变化。结果:目的基因成功转染并在肿瘤组织中有效表达。转基因组与非转移基因组肝脏转移率分别为26.7%(4/15)、100.0%(15/15),2组比较差异有统计学意义(P<0.05),肝脏转移结节数分别为0.60±0.83、2.40±0.99(P<0.001)。镜下转基因组肝内转移癌细胞变性,染色体消失,可见凋亡小体,而非转基因组未见明显变化。结论:原位基因治疗联合前药热化疗对裸鼠结肠癌肝转移有明显抑制作用。

Objective： To investigate the treatment effect of cytosine deaminase gene combined 5-FC thermochemotherapy for colon cancer liver metastasis in Nude Mice. Methods： The models of liver metastasis of colon cancer had been established by injecting human colon cancer cells Lovo into portal vein in 30 BALB/c nude mice,which were divided into transgenic group and non-transgenic group randomly. Transgenic group：virs supernatant was injected into abdominal cavity（0.2 mL/d,for 5 days）; nontransgen c group： sodium chloride was injected into abdominal cavity（0.2mL/d,for 5days）.In both groups 42℃5-FC was injected into the abdominal cavity（500mg·kg^-1·d^-1,for 21days）.After treatment,all mice were sacrificed ,and the PCR and RT-PCR were paerformed to detect the expression of targeted gene in carcinoma.The pathology changes of tumor were observed.Resultes：The targeted gene were detected in the rumor tissues.The rates of liver metastasis tumors were 26.7%（4/15）,100.0%（15/15） in transgenic group and non-transgenic group respectively（P〈0.05）,the number of liver metastasis nodules were 0.60±.83,2.40±0.99respectively （P〈0.001）.In transgenic group,liver metastasis cell denatured,and chromosome disappeared.Conclusions：Cytosine deaminase gene combined 5-FC thermochemotherapy has and obvious inhibiting effect on the growth of liver metastasis of colon cancer in nude mice.