裸鼠原位肝癌移植模型的建立及其生物学观察

目的探讨采用Hep G2细胞株建立裸鼠人肝癌原位模型的方法,评价其生物学特性。方法取人肝癌细胞株Hep G2制备皮下移植瘤,然后将瘤块植入裸鼠肝左叶,建立原位移植模型,观察荷瘤鼠生存状态。裸鼠全身衰竭时处死、取瘤,HE染色及免疫组化法观察瘤组织的病理特点、血管生长及肿瘤细胞凋亡情况。结果种瘤成功率90%,荷瘤鼠中位生存期76.7d,肿瘤直径(16.8±2.2)mm,体积(965.3±333.3)mm^3,重量(1.1±0.4)g。瘤组织早期具有旺盛的增殖能力,晚期癌组织中Caspase3、TUNEL呈强阳性表达。结论由原位移植法建立的裸鼠人肝癌模型,造模时间短、成瘤率高,生物学行为与临床肝癌相似,是基因药物筛选研究的理想模型。

聚合人脐带血红蛋白氧载体增强乳腺癌裸鼠皮下移植瘤化疗敏感性的初步研究

目的探讨聚合人脐带血红蛋白氧载体(PolyCHb)对人乳腺癌MCF-7细胞裸鼠皮下移植瘤化疗敏感性的影响及其机制。方法收集处于指数生长期的MCF-7细胞,制成密^度为5×10^(7)个/mL的悬浮细胞,按0.2 mL/只接种于18只BALB/c-nu裸鼠右肢皮下,建立裸鼠移植瘤模型,待肿瘤体积达到100 mm^(3)左右时,随机均分为化疗组:多柔比星(DOX)5 mg·kg^(-1),1次/周;化疗+PolyCHb治疗组:除给药(DOX)(同化疗组)外,PolyCHb 600 mg·kg^(-1),3次/周;对照组:生理盐水90 mg·kg^(-1),1次/周;均为经尾静脉连续注射4周。自注射当天(d0)起,每3 d 1次测量各组裸鼠肿瘤体积,据此绘制其各自(组)肿瘤生长曲线。38 d结束肿瘤生长观察,剥瘤并称取瘤重,计算抑瘤率;HE染色观察肿瘤组织病理变化,免疫组化检测肿瘤组织中HIF-1α表达,采用TUNEL法检测肿瘤细胞凋亡情况,荧光染色测定各组肿瘤组织活性氧(ROS)的含量。结果化疗+PolyCHb组、化疗组和对照组至d38时的肿瘤体积(mm^(3))分别为:196.35±103.45 vs 316.29±62.88 vs 519.42±177.33(P<0.05);化疗+PolyCHb组与化疗组抑瘤率(%)分别为62.20 vs 39.11;HE染色和TUNEL检测:化疗+PolyCHb组肿瘤组织生长区域细胞坏死和凋亡增多;免疫组化:化疗+PolyCHb组HIF-1α表达水平降低;荧光染色:化疗+PolyCHb组[活性氧(ROS)]含量增多。结论PolyCHb增加了乳腺癌裸鼠皮下移植瘤化疗的敏感性,其作用机制可能与其提高肿瘤组织内ROS含量,促进肿瘤细胞的凋亡有关。

OB glue paste technique for establishing nude mouse human gastric cancer orthotopic transplantation models

AIM： To establish nude mouse human gastric cancer orthotopic transplantation models using OB glue paste technique. METHODS： Using OB glue paste technique, orthtopic transplantation models were established by implanting SGC-7901 and NKN-45 human gastric cancer cell strains into the gastric wall of nude mice. Biological features, growth of the implanted tumors, the success rate of transplantation and the rate of auto-metastasis of the two models were observed. RESULTS： The success rates of orthotopic transplanration of the two models were 94.20% and 96%. The rates of hepatic metastasis, pulmonary metastasis, peritoneal metastasis, lymphocytic metastasis and splenic metastasis were 42.13% and 94.20%, 48.43% and 57.97%, 30.83% and 36.96%, 67.30% and 84.06%, and 59.75% and 10.53%, respectively. The occurrence of ascites was 47.80% and 36.96%. CONCLUSION： OB glue paste technique is easy to follow. The biological behaviors of the nude mouse human gastric cancer orthotopic transplantation models established with this technique are similar to the natural processes of growth and metastasis of human gastric cancer, and, therefore, can be used as an ideal model for experimental research of proliferative metastasis of tumors.

Establishment of orthotopic transplantation model of human bladder cancer and detection by MRI

Objective: To establish an orthotopic bladder cancer model bearing human bladder cancer for experimental research, and monitor tumor progression by magnetic resonance imaging (MRI). Methods: The mucosa was mechanically damaged transurethrally under direct vision, and then human bladder cancer cell line T24 was inoculated into the bladders of BALB/c nude mice to establish orthotopic bladder cancer model. To find a suitable concentration of Gd-DTPA for this re- search. MRI was performed weekly to assess tumor growth, using Gd-DTPA as contrast agent. The pathologic morphology of the bladders and other specimens were observed with HE stain. Results: All the 25 mice developed bladder cancer after inoculation. The best concentration of Gd-DTPA was 1.408 mg/mL. On MRI, no change in the bladders was observed on day 7 after inoculation, filling defect in the bladders, accordant to actual tumor size, was detected on days 14, 21 and 28. Pathologic examination showed that tumor grew in the mucosa or superficial muscle of bladder on day 7, confined in muscle layer on days 14–28, and invaded serosa on day 35. Conclusion: Transurethrally damaged bladder mucosa under direct vision and instilled bladder cancer cell T24, we successfully established an orthotopic bladder cancer model. Tumor growth simulated the progression of human bladder cancer approximately. MRI was a reliable way for dynamic detection of murine orthotopic bladder tumor.

Establishment of orthotopic impact/metastasis model of human ovary cancer in nude mice

Objective: To establish a patient-like human ovary carcinoma /spontaneous metastasis model using orthotopic transplanation of histologically intact tumor tissue. Methods: An highly metastatic ovarian tumor line (HO-8910PM: Human serum carcinoma of the ovary )previously grown substaneously was transplanted into the ovicapsule using microsurgery technique .Histologically intact human ovary tumor pieces gained from implantation site were passaged between ovicapsules for four generations. Results: All mice developed ovary tumors and the metastatic rates were about 75%. The tumors only metastasized to liver but no other organs. The earliest appearance of metastasis was 14 d and the average survival period was 20.7±4.89 d.The microscopic appearance of the metastases was similar to the tumor observed in the substaneous xenografts and orthotopically transplanted. Chromosomes analysis exhibited the feature of human carcinoma and retained genetic stability during the processes of passage. Conclusion: Orthotopic implanation provides a suitable micro-enviroment in which ovarian cancer can express its intrinsic clinically-relevant properties. This approach is relevant to the spontaneous development of ovarian cancer and is thought to be a useful model for studies of metastatic mechanism and therapy for ovary cancer.