人宫颈癌SiHa细胞荷瘤裸鼠模型的建立及鉴定

Establishment and identification of human cervical cancer Si Ha cells tumor bearing model in nude mice

目的:建立人宫颈癌SiHa细胞裸小鼠体表荷瘤模型,并对其成瘤特性进行分析和鉴定。方法:将5×10~6/mL、1×10~7/mL及5×10~7/mL 3个浓度的SiHa细胞悬液分别在3组裸小鼠背部右腋近上肢皮下接种,建立人宫颈癌裸小鼠体表荷瘤模型,观察荷瘤裸鼠的成瘤率并测量肿瘤的大小,5周后取肿瘤组织经病理学方法观察其组织学特性,并采用PCR及免疫组织化学方法对肿瘤组织的HPV16E7 DNA表达进行检测。结果:3组裸小鼠接种SiHa细胞的成瘤率均为100%,5周后肿瘤大小分别达到(77.29±28.34)mm^3、(178.91±61.55)mm^3和(305.11±12.62)mm^3。3个浓度组间肿瘤体积大小差异有统计学意义(P<0.001)。病理学大体观察可见SiHa肿瘤的生长均以局部浸润为主;组织切片观察新生肿瘤细胞特点呈现体积大、核大深染及易见核分裂相的病理性特征;SiHa新生肿瘤组织经PCR检测证实了HPV16E7 DNA阳性,并经免疫组织化学检测证实了HPV16E7蛋白表达阳性。结论:成功建立了SiHa细胞荷瘤裸鼠模型,为人宫颈癌肿瘤靶向治疗及生物学特性研究提供了理想的动物模型。

Objective： To establish human cervical cancer SiHa cells tumor bearing model on the body sur-face area in nude mice, and analyze its tumor characteristics. Methods： Three groups nude mice were subcutane- ous inoculated with 5 × 106/mL, 1 ×107/mL and 5 × 107/mL concentrations of cervical cancer SiHa cells in the right axillary close to upper limbs, to establish human cervical cancer SiHa cells tumor bearing model in nude mice. The tumor of nude mice was observed and the size of tumor was measured. The histological characteristics of tumor tissue was observed by pathological method after 5 weeks. HPV16E7 DNA of tumor tissue was comfirmed by PCR assay, and the expression of HPV16 E7 protein in tumor tissue was detected by immunohistochemistry method. Results： The tumor rate of 3 groups of nude mice inoculation with SiHa cell line was 100%. Their tumor sizes reached up （77.29±28.34） mm3, （178.91±61.55） mm3 and （305.11±12.62） mm3 after 5 weeks, and there was a significant difference among 3 groups （P〈0.001）. Pathology observation showed the growth of SiHa cell tumors were mainly in local infiltration. Histopathological of slice observation showed the new SiHa cells tumor tissue appeared pathological characteristics of large volume, large nucleus, nucleus deep dyeing and the mitotic phase. HPV16E7 DNA and the protein expression were positive comfirmed by PCR and immunohistochemistry detec- tion respectively in tumor tissue. Conclusion： Human cervical cancer SiHa cells tumor bearing model in nude mice was established successfully, and provide an ideal animal model for human cervical cancer tumor targeted therapy and biological research.