卵巢癌SKOV3细胞卵巢原位种植模型的建立及鉴定

The identification and the establishment of the orthotopic transplantation mouse model by ovarian cancer cells SKOV3

目的记录通过原位凝胶种植法荷瘤后的NOD-SCID鼠的肿瘤形成过程、肿瘤转移情况及腹水形成时间。方法以绿色荧光蛋白（GFP）转染SKOV3细胞。将转染后的SKOV3细胞以一定浓度种植于裸鼠及NOD-SCID鼠体内,根据小鼠成瘤情况及死亡时间判断最佳种植部位和最适种植细胞数。以最适部位和细胞数再次种植20只NOD-SCID鼠,每周记录肿瘤大小和腹水生成情况及肿瘤转移情况。通过ELISA法检测荷瘤鼠血清中CA125水平,通过免疫组化检测肿瘤组织中BRCA1、BRCA2及Ki67的表达情况。结果原位种植为凝胶种植法最佳种植部位,4×10~4/个SKOV3细胞为最适种植种植浓度。荷瘤鼠血清CA125水平明显升高,且与荷瘤时间成正比,肿瘤组织中BRCA1、BRCA2、Ki67的表达呈阳性。结论通过SKOV3细胞原位凝胶种植法建立的卵巢癌动物模型具有成瘤时间短,发展过程与人卵巢原发肿瘤更为接近等特点,可为卵巢癌新治疗方法的研究提供较好的动物模型。

Objective To records the detail progression of the tumor,the metastasis and the ascites in the ovarian mouse model developed by orthotopic collagen implantation in NOD-SCID mouse. Methods The ovarian cancer cell line SKOV3 was transfected with Green fluorescent protein. Preexperimentals were processed on nude and NOD / SCID mice in order to estimate the optimal planting area and the optimum planting cell count. The best planting area and the appropriate concentration of cells were chosen. According to the results,anther 20 NOD / SCID was transplanted. The detection of the turmor size,the metastase and the ascite was performed weekly. The expressions of BRCA1,BRCA2 and Ki67 were also identified by immunohistochemistry. The serum CA125 level was determined by ELISA. Results The optimal planting area was ovray and the concentration was 4 × 104 cells. The serum CA125 level increased significantly compared with the healthy mice. BRCA1,BRCA2,P53 and Ki67 showed positive in the tumor tissues by immunohistochemistry. Conclusion The characteristics of the orthotopic transplantation mouse model of ovarian cancer built by collagen implantation is rapid tumorigenesis and it is superior to reflecting the biological behavior of ovarian cancer. This humanized mouse model may be used as preclinical tool to investigate ovarian cancer.