宫颈癌移植瘤大鼠动物模型的建立

Establishment of cervical cancer transplanted carcinoma models in rats

目的:应用瘤细胞悬液移植法和静脉注射方法将人宫颈癌HELA细胞接种于具有正常免疫功能的大鼠体内建立人的宫颈癌移植瘤模型,观察成瘤时间,肿瘤进行性生长情况和其他实质性器官转移情况。方法:实验于2004-10/2005-06在中国医科大学盛京医院实验动物部及超声科进行。①实验动物:成年雌性Wistar大鼠35只,随机分为皮下注射组10只、腹腔注射组10只、静脉注射组10只、阴性对照组5只。②实验方法:取对数生长期的人宫颈癌HELA细胞。皮下注射组接种2.5×1010L-1细胞悬液于大鼠右后肢及背部皮下。腹腔注射组接种2.5×109L-1细胞悬液于腹腔内。静脉注射组由大鼠尾静脉注入2.5×108L-1的细胞悬液。没有进行免疫抑制的5只大鼠做阴性对照。③实验评估:接种后观察成瘤率、肿瘤生长及播散情况及动物生存期,行超声检查观察肿瘤的二维灰阶表现和肿瘤的血供情况。结果:①皮下注射组7只成瘤,腹腔注射组8只成瘤,静脉注射组10只,在肿瘤生长过程中3只死亡,其余7只全部成瘤。总体成瘤率67%。腹腔种植组及静脉注射组的肿瘤较早出现腹腔脏器转移。皮下注射组成瘤时间较晚,约种植后40d左右。②3组平均生存时间分别为静脉注射组62.5d,腹腔注射组78.9d和皮下注射组84.4d,差异有显著性意义(P<0.01)。③超声检查皮下生长的肿瘤组织呈低回声,椭圆形,内部回声均匀,边界清晰,彩色多谱勒扫查未见彩色血流信号显示。转移较明显的腹腔种植组及静脉注射组大鼠超声检查可见,2周后,肝区内结节表现为椭圆形或不规则形的低或略高回声,内部质地欠均匀,边缘尚清晰,但不是很规则。4周左右于肝区内见异常低回声结节或强回声结节,大小约0.4cm×0.5cm。肿瘤内部未见明显血流信号,外周可见少许滋养血管。结论:人宫颈癌HELA细胞接种于具有正常免疫功能的大鼠体内建立人的宫颈癌移植瘤模型,具有与人宫颈肿瘤的组织学形态及生物学行为相似的特点,且移植瘤的生长状况、回声类型及血供变化大体符合宫颈癌的一般超声表现特点。

AIM： To establish models of cervical cancer transplanted carcinoma in immunocompetent rat by seeding human cervical carcinoma （HELA） cells carcinoma using cell suspension and intravenous injection, and to observe carcinoma formation time, progressive growth and parenchymatous organ metastasis. METHODS： The experiment was performed at Department of Experimental Animal and Department of Ultrasound, Shengjing Hospital of China Medical University from October 2004 to June 2005. Thirty-five adult female Wistar rats were randomly divided into 4 groups： in subcutaneous injection group （n=10）, 2.5 × 10^10 L^-1 HELA cells was subcutaneously injected into right hind limb and back; in intraperitoneal injection group （n=10）, 2.5 × 10^9 L^-1 HELA cells were intraperitoneally injected; in intravenous injection group （n=10）, 2.5 × 10^8 L^-1 HELA cells were injected. Five rats without immunosuppressive intervention served as negative control. Tumor formation rate, tumor growth and animal survival were observed. The 2-dimensional gray-scale and blood supply of tumor were observed by ultrasonography. RESULTS： Tumor formation was found in 7 rats of subcutaneous injection group, 8 of intraperitoneal injection group and 10 of intravenous injection group. During tumor growth, 3 rats die. Total tumor formation rate was 67%. Tumor metastasis of abdominal cavity organ was early found in intraperitoneal injection group and intravenous injection group. Tumor formation was observed in subcutaneous injection group about at 40 days. Mean survival time of intravenous injection group, intraperitoneal injection group and subcutaneous injection group was 62.5, 78.9 and 84.4 days, respectively （P 〈 0.01）. Ultrasonographic observation showed that subcutaneous tumor tissue was at low echo-level, and oval-shaped. Internal echo was even with clear border. No color blood flow signal was found in color Doppler scanning. Notable tumor metastasis was found in intravenous injection group and intraperitoneal injection group. Two weeks after injection, nodules of liver presented oval or irregular low or high level echo; the echo was even and the border was clear but not disorderly. At about 4 weeks, abnormal hypoecho or hyperecho nodules were observed in liver, about 0.4 cm × 0.5 cm. No obvious blood flow signal was found in tumor, but few nutrient vessels were found in the outside. CONCLUSION： This animal model has similar histomorphological and biological characteristics as human cervical carcinoma. Moreover, metastasis tumor growth, echo patterns and blood supply are nearly in accordance with general ultrasonic manifestation of cervical carcinoma.