【作者】 王朝霞；

【导师】 李莉；

【作者基本信息】 山西医科大学 ， 妇产科学， 2010， 硕士

【摘要】 目的：探讨基因重组质粒pDC316-hIL-24对裸鼠人宫颈癌模型的肿瘤生长及凋亡的作用。方法：于5周龄裸鼠腋窝外侧皮下注射1×107/mlHela细胞悬液100μl(约1×106),将构建成功的人宫颈癌Hela细胞裸鼠模型,随机分为3组,分别于瘤体内注射基因重组质粒pDC3.0-hIL-24,空质粒pDC3.0以及磷酸盐缓冲液(PBS),比较干预后移植瘤的体积及抑瘤率,应用PCR法测定移植瘤中hIL-24的表达,HE染色观察各组肿瘤组织形态学变化,免疫组化方法研究肿瘤细胞凋亡情况。结果：经转染IL-24基因已成功转染入肿瘤细胞并在肿瘤细胞中已成功表达,至实验结束时,对照组和空质粒组移植瘤体积明显大于IL-24基因重组质粒组(F=63.395,P＜0.05)；IL-24重组质粒组的平均抑瘤率明显高于空质粒组(t=130.920,P＜0.01)；病理学观察见IL-24重组质粒组宫颈癌细胞有局灶性坏死,细胞变大,胞质疏松,核深染,可见核固缩、核溶解、核碎裂,染色质成团状；免疫组化结果显示,与其余两组相比,基因重组质粒组原癌基因Bcl-2表达降低(F=6.164,P＜0.05),而其同源蛋白Bax表达明显高于其他两组(F=58.426,P＜0.05)。结论：基因重组质粒pDC316-hIL-24对宫颈癌裸鼠模型肿瘤生长具有明显的抑制及促进凋亡作用,IL-24可能通过激活细胞凋亡/抗凋亡通路而诱导肿瘤细胞核固缩碎裂、细胞凋亡,而且IL-24抗肿瘤作用对动物生活能力及质量无明显不良影响。更多还原

【Abstract】 MethodCervical cancer Hela cells were injected in a 106concentration into the central lateral subcutaneous of the left armpit of nude mice of 5 weeks age. Construction of nude mice model of human cervical cancer. The nude mice were randomly divided into 3 groups, and recombinant plasmids or empty plasmids as well as PBS were injected into transplanted tumor to interfere with the growth of transplanted tumor. Comparison of tumor volume and inhibitory rate after the intervention, determination of hIL-24 expression in transplanted tumor by PCR, observation of morphological changes of tumor tissues in each group by HE staining and as well as immunohistochemistry was employed for investigating tumor cell apoptosis in each group.ResultAfter transfection IL-24 gene was successfully transfected into tumor cells and has been successfully expressed in tumor cells. To the end of the experiment, the control group and blank plasmid group tumor volume was significantly greater than IL-24 gene plasmid group (F=63.395, P<0.05); The average inhibition rate of IL-24 recombinant plasmid group was significantly higher than empty plasmid group (t=130.920, P<0.01); Pathological observation shows in plasmid IL-24 group cervical cancer cells have focal necrosis and cells become large, and loose cytoplasm, nucleus deeply stained, pyknosis, nuclear dissolution, nuclear fragmentation, chromatin lumps also can be found; Immunohistochemistry showed that, compared with the other two groups, recombinant plasmid group proto-oncogene Bcl-2 expression was decreased (F=6.164, P<0.05), but its homologous protein Bax expression was significantly increased (F=58.426, P<0.05).ConclusionRecombinant plasmid pDC316-hIL-24 has significant inhibition function on human cervical cancer nude mice model tumor growth. IL-24 may activate apoptosis/ anti-apoptosis pathway to induce tumor cell nuclear condensation, fragmentation and apoptosis, and anti-tumor function of IL-24 had no obvious adverse effects on the quality of animals’life.更多还原