人卵巢癌裸鼠卵巢原位移植瘤模型的构建

Orthotopic construction of human ovarian carcinoma xenograft model in nude mice

为探讨人卵巢癌进展机制的相关研究,构建适宜的体内动物模型。采用慢病毒转染技术,构建稳定过表达绿色荧光报告基因(GFP)的人EOC细胞模型:SKOV3(人卵巢浆液性腺癌细胞株)和ES-2(人卵巢透明细胞癌细胞株);异氟烷吸入麻醉4-6周龄BALB/C(nu/nu)雌性裸鼠,从裸鼠背部正中纵切口进入腹腔,显微多点注射法分别将细胞模型悬液,移植于裸鼠的单侧卵巢,每组6只裸鼠;Night OWL活体成像系统动态观察裸鼠移植瘤生长和转移情况;移植术后4-6周解剖裸鼠,取卵巢和转移瘤组织行HE染色。结果显示,携带GFP的EOC细胞模型SKOV3和ES-2,均可成功构建裸鼠卵巢原位移植瘤模型,裸鼠未发生手术原因死亡,成功率均100%;活体成像系统通过检测GFP的荧光强度和范围,能够动态监测裸鼠移植瘤生长情况;SKOV3细胞模型构建的卵巢癌和转移瘤组织,HE染色观察病理特征符合人卵巢浆液性腺癌;ES-2细胞模型构建的卵巢癌和转移瘤组织,HE染色观察病理特征符合人卵巢透明细胞癌。由此可知,异氟烷吸入麻醉、从背部正中纵切口进入腹腔、显微多点注射法移植携带GFP的EOC细胞模型,能成功构建可动态监测的裸鼠卵巢原位移植瘤动物模型;该动物模型与人卵巢癌生物学特征类似,适宜作为研究人卵巢癌演变进展的体内模型。

To build a suitable animal ovarian cancer model in vivo, so as to study the mechanism of its progression.Lentivirus transfection technology was used to build a green fluorescent protein（GFP） marked EOC cells model： SKOV3（human ovarian serous carcinoma cell lines） and ES-2（human ovarian clear cell carcinoma cell lines）. After isoflurane anesthesia, 4-6 week old BALB/C（nu/nu） nude female, back into the abdominal cavity from the median longitudinal incision. Micro multipoint injection method was used to transplant suspension cell model in one side ovary; Night OWL vivo imaging system was used to dynamically observe the growth and metastasis of xenografts in nude mice. Nude mice were dissected at 4-6 weeks after transplantation, the ovarian and metastatic tissues were stained with HE. Both SKOV3 and ES-2 GFP positive cell model was successfully constructed with ovarian orthotopic xenograft. nude mice did not lead to surgical death, the success rate of orthotopic transplanted tumor model in nude mice was 100%. The in vivo imaging system can dynamically monitor the growth of a transplanted tumor in nude mice by detecting the fluorescence intensity and range of GFP. SKOV3 cell model was built-in ovarian canceer and metastatic tissues, HE staining pathological features consistent with human ovarian serous adenocarcinoma; ES-2 cell model was built-in ovarian cancer and metastatic tissues, HE staining pathological features consistent with human ovarian clear cell carcinoma. it can be concluded that inhalation of isoflurane anesthesia, middle of the back vertical incision into the abdominal cavity, micro multipoint injection of GFP-carrying EOC cell model, can be successfully constructed dynamic observation of ovarian orthotopic transplantation tumor in nude mice animal models. This animal model is similar to the biological characteristics of human ovarian cancer and is suitable as an in vivo model to study the development of human ovarian cancer.