Anti-tumor activities and apoptosis-regulated mechanisms of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice

AIM: To investigate anti-tumor activities and apoptosis-regulated mechanisms of bufalin in the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice.METHODS: BEL-7402 cells of human hepatocellular carcinoma were inoculated to form subcutaneous tumors, and were implanted into the liver to establish orthotopic transplantation tumor models of human hepatocellular carcinoma in nude mice. Seventy-five animals were randomized divided into five groups (n = 15). Bufalin was injected intraperitoneally into three groups at doses of 1.5 mg/kg (BF1), 1 mg/kg (BF2) and 0.5 mg/kg (BF3) for d 15-24, respectively. The NS group was injected an equal volume of saline as above and adriamycin was injected intraperitoneally into the ADM group at a dose of 8.0 mg/kg for d 15. Ten mice in each group were killed at d 25 and the survival time in each group was calculated. We also observed the morphologic alterations in the myocardium, brain, liver, kidney and tumor tissues by pathology and electron microscopy, measured the apoptotic rate by TUNEL staining method, and detected the expression of apoptosis-regulated genes bcl-2 and bax by immunohistochemical staining and RT-PCR in tumor tissues. RESULTS: The tumor volumes in each group of bufalin were reduced significantly (35.21 ± 12.51 vs 170.39 ± 25.29; 49.83 ± 11.46 vs 170.39 ± 25.29; 83.99 ± 24.63 vs 170.39 ± 25.29, P < 0.01, respectively), and the survival times were prolonged in group BF1-2 (31.8 ± 4.2 vs 23.4 ± 2.1 and 29.4 ± 3.4 vs 23.4 ± 2.1, P < 0.05, respectively), and necrosis was mainly in severe or moderate degree in group BF1-2. No morphologicalchanges were detected in the myocardium, brain, liver and kidney tissues. Apoptotic characteristics could be seen in group BF1-2. The positive rates of bcl-2 and bax protein expression of each group by immunohistochemical staining were 10.0%, 10.0%, 20.0%, 10.0% and 20.0%; 90.0%, 80.0%, 80.0%, 40.0% and 30.0%, respectively. Loss of expression of bcl-2 mRNA in each group was to be found and the density of bax mRNA was increased progressively with increase of dose of bufalin by RT-PCR. CONCLUSION: Bufalin has significant anti-tumor activities in the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice with no marked toxicity and was able to induce apoptosis of transplanted tumor cells. This apoptosis may be mediated mainly via up-regulating the expression of apoptosis-regulated gene bax, which may be involved in its anti-tumor mechanism of bufalin.

Anti-tumor effect of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice

Objective: To investigate anti-tumor effect of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice. Methods: BEL-7402 cells of human hepatocellular carcinoma were inoculated to form subcutaneous tumors in nude mice by subcutaneous injection. Then the subcutaneous tumors were implanted into the liver of nude mice, and the orthotopic transplantation tumor models of human hepatocellular carcinoma were established. Seventy-five models were randomized into 5 groups ( n = 15) . Bufalin was injected intraperitoneally into the 3 groups at dose of 1.5,1 and 0.5 mg/kg for day 15 - 24, respectively. NS group were injected equal volume saline as above and adriamycin were injected intraperitoneally into ADM group at dose of 8.0 mg/kg for day 15. Ten mice in each group were killed at day 25 and detected on morphological and ultrastructural changes in myocardium, brain, liver, kidney and tumor tissues by pathology and electron microscope. The survival time in each group were observed. Results: The tumor volumes in each group of bufalin were reduced significantly compared with NS group (P < 0.01), the survival time were prolonged in group Bu 1 and Bu 2 compared with NS group ( P < 0.05), and tumor tissues were mainly necrosis in severe or moderate degree in Bu 1, Bu 2 groups, and mild degree or moderate degree in Bu 3 group. No morphological changes were detected in myocardium, brain, liver and kidney tissues, respectively. Apoptotic characteristics could be seen in tumor tissues of group Bu 1 and group Bu 2. Conclusion: Bufalin has significant anti-tumor effects on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice without marked toxicity. To guide cell apoptosis may be one of its anti-tumor mechanism of bufalin.

Establishment of an orthotopic transplantation tumor model of hepatocellular carcinoma in mice

AIM:To improve the outcome of orthotopic transplantation in a mouse model,we used an absorbable gelatin sponge(AGS) in nude mice to establish an orthotopic implantation tumor model.METHODS:MHCC-97L hepatocellular carcinoma(HCC)cells stably expressing the luciferase gene were injected into the subcutaneous region of nude mice.One week later,the ectopic tumors were harvested and transplanted into the left liver lobe of nude mice.The AGS was used to establish the nude mouse orthotopic implantation tumor model.The tumor suppressor gene,paired box gene 5(PAX5),which is a tumor suppressor in HCC,was transfected into HCC cells to validate the model.Tumor growth was measured by bioluminescence imaging technology.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) and histopathology were used to confirm the tumorigenicity of the implanted tumor from the MHCC-97L cell line.RESULTS:We successfully developed an orthotopic transplantation tumor model in nude mice with the use of an AGS.The success rate of tumor transplantation was improved from 60% in the control group to 100% in the experimental group using AGS.The detection of fluorescent signals showed that tumors grew in all live nude mice.The mice were divided into 3 groups:AGS-,AGS+/PAX5-and AGS+/PAX5 +.Tumor size was significantly smaller in PAX5 transfected nude mice compared to control mice(P < 0.0001).These fluorescent signal results were consistent with observations made during surgery.Pathologic examination further confirmed that the tissues from the ectopic tumor were HCC.Results from RT-PCR proved that the HCC originated from MHCC-97L cells.CONCLUSION:Using an AGS is a convenient and efficient way of establishing an indirect orthotopic liver transplantation tumor model with a high success rate.

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.

Orthotopical transplantation of human renal carcinoma tissue into nude mice and the establishment of a high metastatic cell line MRCC

Objective To establish a SOI model of human renal carcinoma and a high metastatic cell subline. Methods A human renal cell line RCC-9863 has been established by inoculating a human renal tumor tissue into nude mice s. c.. When RCC-9863 passaged for 20 times, the tissue from the same xemotransplant tumor were used to construct SOI model. Cultured the metastatic tissue in vitro, the tumor cell suspension was then injected orthotopically, The metastatic tissue obtained underwent the same procedure again. At last, the metastatic tumor was cultured in vitro and cloned. Results 15 days later, a tumor mass sized 1. 7 cm × 0. 6 cm in the nude mouse’s renal parenchyma was grown which lobulated, rude, and with multiply blood vessels and 55 days later later the mouse became moribund and metastases in the lungs were formed. The transplanted renal tumor in the SOI model grew fast and invasively and metastasized to lungs, lymphatic node and liver. A subline, MRCC, with metastatic ability to the lung was selected.

Orthotopic transplantation model of human gastrointestinal cancer and detection of micrometastases

AIM: To establish a relevant animal model of human gastrointestinal cancer, which can be used for repetitive investigations, so as to improve our understanding and management of carcinogenesis and cancer metastasis. METHODS: Intact tissues of human colorectal and pancreatic cancers were transplanted in nude mice. The biological characteristics of the original and the corresponding transplanted tumors were investigated by HE staining, PAS staining and immunostaining. The metastases in the livers and lungs of nude mice were investigated by immunostaining with biotinylated mab KL-1 and by RT-PCR using CK20 specific primers. RESULTS: There were totally 9 of 16 surgical specimens growing in nude mice subcutaneously and/or orthotopically (4 of 6 colorectal and 5 of 10 pancreatic cancer). Tumor cell content of the specimens and freezing of tissue specimens are important factors influencing the growth of transplanted tumor. In the group of fresh tumor tissues with greater than 50% tumor cell content, the success rate of the transplantation was 100% (3 cases of pancreatic cancer and 3 cases of colorectal cancer). The orthotopically trans-planted tumors resemble the original tumor morphologically and biologically, including TAA expression such as CEA by immunohistochemistry, and CEA level in the serum of mice. Ki-67 labeling index and the expression of TAA especially K-ras, 17-1A and RA-96, are associated with the potential of tumor growth in nude mice. Micrometastases in the lungs and livers of tumor bearing mice can be detected by immunostaining with biotinylated mab KL-1 and CK20-specific RT-PCR. CONCLUSION: An orthotopic transplantation model for human colon and pancreatic cancer in nude mice has been set up. We have also established sensitive detection methods with CK-immunohistochemistry and CK20-RT-PCR to study xenotransplanted human cancer and its metastatic cancer cells in the liver and lung of nude mice. This study may be helpful in understanding the mechanism of cancer metastasis and in developing new diagnostic methods and therapeutic strategies for metastases including micrometastases.

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.

人肝肿瘤细胞的裸小鼠原位癌建模条件优化及评价

目的优化通过注射人肝肿瘤细胞株构建原位癌裸小鼠模型的条件,并探索适宜的给药治疗时间。方法选用稳定表达萤光素酶报告基因(LUC)的人肝细胞癌Hep3B与肝母细胞瘤HepG2细胞株,使用小动物活体成像系统分析萤光素酶发光强度与肝肿瘤细胞数量之间的线性相关性,验证人源肝肿瘤细胞的发光效率。在5周龄雌性BALB/c裸小鼠的肝叶原位接种不同浓度(8×10^(6)、2.4×10^(7)、7.2×10^(7)个/mL)、不同重悬介质(PBS、Matrigel)的人肝肿瘤细胞悬液HepG2-LUC和Hep3B-LUC(共12组,每组7只),分别构建人肝肿瘤裸小鼠原位癌模型。每7 d为1个周期记录各组小鼠体重,用小动物活体成像系统定期监测原位肿瘤的生长过程,观察肿瘤生长趋势。接种肿瘤细胞后第35天剖取小鼠肝脏,制备病理切片,进行苏木精-伊红(hematoxylin and eosin,HE)染色观察组织病理学变化。结果两种人肝肿瘤细胞株的发光强度均与细胞数量呈正相关(R^(2)=0.9831,R^(2)=0.9705),适宜用于原位癌模型的构建。HepG2-LUC高浓度组,HepG2-LUC+Matrigel低、中、高浓度组,Hep3B-LUC中、高浓度组与Hep3B-LUC+Matrigel低、中、高浓度组均成功造模。HepG2-LUC+Matrigel高浓度组较低浓度与中浓度组小鼠的体重显著下降(P<0.05),Hep3B-LUC+Matrigel高浓度组较低浓度与中浓度组小鼠的体重也显著下降(P<0.05)。成功造模组小鼠的荧光发光强度随时间呈指数型增长(R^(2)>0.9500),且在移植后14 d发光强度至少可达到1.0×10^(7) p/(s·cm^(2)·sr)。HepG2-LUC低、中浓度组和Hep3B-LUC低浓度组小鼠肝脏未见明显的病理学变化,其余组肝脏肿瘤和肝细胞病变明显。结论对于HepG2-LUC细胞株,推荐肝叶原位注射2.4×10^(7)个/mL(50μL)且与Matrigel重悬的混合细胞液体造模,并于造模后第7天给药或采取预后措施;而对于Hep3B-LUC细胞株,推荐肝叶原位注射7.2×10^(7)个/mL(50μL)(不与Matrigel重悬混合)造模,并于造模后的第14天给药或采取预后措施。

完整瘤组织块原位移植法建立人成骨肉瘤转移模型

目的：建立人成骨肉瘤裸鼠转移模型。方法：采用完整瘤组织块原位移植法获得裸鼠自发性肺转移模型，并对转移模型进行影像学检查、组织学检查及 PCR同源性分析。 结果： 建立裸鼠体内人成骨肉瘤转移模型，其自发性肺转移率达到 100％，肝转移率达 30％。影像学检查及组织学检查均与人成骨肉瘤临床情况相似， PCR同源性分析证实裸鼠肺部转移灶为人成骨肉瘤细胞。结论：裸鼠体内人成骨肉瘤自发性转移模型模拟了临床发病过程，对人成骨肉瘤的治疗、局部肿瘤的生长及转移机制的研究提供了更为理想的模型。

蚯蚓纤溶酶抗肝癌转移作用的实验研究

目的探讨蚯蚓纤溶酶(earthworm fibrinolytic enzyme,EFE)对人肝癌细胞转移的影响。方法建立裸鼠人肝癌高转移原位移植瘤模型,造模后7d将裸鼠随机分为模型对照组和EFE高、低剂量组,共3组,每组7只,每天分别给予生理盐水和1600,800uku/kgEFE灌胃,连续30d。实验结束时完整剥除种植瘤并称重;肉眼直接观察种植瘤的肝内播散及腹腔种植情况,计算肝内播散率及腹腔种植率;通过病理切片观察肺转移肿瘤灶的数目;采用RT-PCR及Western blot方法检测移植瘤中FAK(focus adhesion kinase)及β1-整合素蛋白的表达。结果与模型对照组相比,EFE低、高剂量组原位种植瘤瘤重减轻(P<0.05或P<0.01);种植瘤肝内播散率和腹腔种植率降低,其中,EFE高剂量组与模型对照组比较,具有显著性差异(P<0.05);肺转移灶数目明显减少,与模型对照组相比,具有显著性差异(P<0.05或P<0.01)。在原位种植瘤和移植瘤中FAK及β1-整合素在mRNA及蛋白水平的表达方面,EFE高、低剂量组的表达均明显下降,与模型对照组比较具有非常显著性差异(P<0.01)。结论EFE具有抗肝癌转移的作用,其机理可能与EFE能够抑制FAK及β1-整合素的表达有关。

人卵巢癌裸鼠移植瘤模型的建立及其生物学特性的实验应用研究

研究用１例人卵巢癌淋巴转移的癌组织直接移植于裸鼠皮下，建成一株人卵巢癌移植瘤动物模型，已传至５经２６代。移植成功率达１００％，平均裸带瘤存活中位数为１０２。肿瘤倍增时间为７．１７天。组织学和超微结构形态证实保持了原人肿瘤的特征，有淋巴结转移行为。人类肿瘤染色体特征。保留了分泌癌胚抗原的能力。具有Ｐ５３癌基因蛋白的异常表达。移植瘤细胞可在体外培养并传至５例。流式细胞仪及显微光光度计检测移植瘤。

可移植性人脑胶质瘤组织裸小鼠脑内移植及其MR显像研究

背景与目的:文献报道的胶质瘤动物原位移植模型,大多数是将细胞悬液立体定向接种于鼠脑内,操作繁杂,耗时长,难以在短时间内完成批量实验。本研究接种可移植性人脑胶质瘤组织于裸小鼠脑内,探讨建立人脑胶质瘤裸小鼠原位移植模型及其MR活体成像的可行性。方法:在套管针内,置入位于裸小鼠皮下生长的可移植性人脑胶质瘤组织2mm3,经微型颅钻钻颅孔后经套管针推入裸小鼠右尾状核内,第30天在配有小鼠专用micro-23微线圈的1.5TMR机上进行头颅扫描和专用软件测量显像的肿瘤体积。继而取全脑作连续冰冻切片,HE染色,在光学显微镜下观察肿瘤病理特征,在体视显微镜目镜下用测微尺测量肿瘤。然后将上述两种方法测得的数据进行统计学分析,评价荷瘤鼠活体MR显像计算肿瘤体积的可行性。结果:经MR扫描的15只颅内接种肿瘤的小鼠,有14只的尾状核区域见到肿瘤显像;在脑切片上,与MR像相同部位见到了肿瘤组织。经脑切片测得的肿瘤体积[(23.19±10.18)mm3]和经MRI测得的肿瘤体积[(23.45±11.64)mm3]比较差异无统计学意义(P>0.05)。肿瘤模型制作成功率为93%(14/15),肿瘤模型MR显像成功率为100%(14/14)。结论:经套管针定量植入胶质瘤组织于裸小鼠尾状核,制作可移植性裸小鼠人脑胶质瘤原位模型,具有操作便捷、省时、便于批量制作、致瘤率高等优点。配有小鼠专用微型线圈的1.5TMR机可用于荷瘤裸小鼠的头颅成像、在活体上对移植瘤进行定位和体积测量等研究。

人结肠癌裸鼠原位移植瘤模型的建立

目的建立人结肠癌裸鼠原位移植瘤模型。方法使用对数生长期的人结肠癌细胞(lovo)在8只裸鼠结肠浆膜至黏膜逐层注射,以完成原位移植瘤模型的制备,同时皮下种植8只裸鼠作为对照组。分别于第4、6、8、12周各组分别处死裸鼠2只,观察原位种植肿瘤的成瘤率、生长情况、转移率和腹水出现率。结果16只裸鼠实验期间无1只死亡,成瘤率为100%,原位种植成瘤率为100%(8/8),区域淋巴结转移率100%(8/8),肝转移率为100%(8/8),肺脏转移率为75.0%(6/8),腹膜转移率为75.0%(6/8),腹水出现率为27.5%(3/8)。皮下种植组未见转移。结论本实验成功建立了人结肠癌裸鼠原位移植瘤模型,该模型的生物学行为与临床病程非常相似,为研究人结肠癌转移机制和干预措施提供了较为理想的动物模型。

绿色荧光蛋白标记的人卵巢癌裸鼠原位移植模型的建立

目的应用绿色荧光蛋白(GFP)基因标记人卵巢癌细胞株HO8910,建立新型裸鼠原位移植瘤模型。方法将处于对数生长期的人卵巢癌细胞株HO8910/GFP2×106个细胞注射于裸鼠腋下,收集瘤块后接种于左侧卵巢包膜下,共6只,术后利用荧光体视显微镜连续观察肿瘤生长情况,4周后处死荷瘤鼠,观察肿瘤生长及转移情况。结果种植的原位移植瘤成瘤率100%,移植后2周可观察到左侧肋脊角处绿色荧光团,并逐渐增大,4周后,可见肿瘤侵及腹膜、网膜、肠管、脾脏、肝脏、子宫、盆腔淋巴结,转移率达66.7%。结论成功建立新型裸鼠原位人卵巢癌的动物模型。

蟾毒灵对裸鼠大肠癌原位移植瘤的抗肿瘤作用及其对凋亡相关基因Bcl-x_L、Bax表达的影响

目的探讨蟾毒灵对裸鼠大肠癌原位移植瘤的抗肿瘤作用及其可能诱导凋亡的机制。方法将60只人结肠癌细胞株HCT-116建立的裸鼠原位移植瘤模型随机分为生理盐水(NS)组、5-Fu组、蟾毒灵低(BL)、中(BM)、高(BH)5组,每组12只,腹腔注射给药持续7d。用药结束后第24天每组分别处死6只裸鼠,完整取出肿瘤,测量肿瘤大小,计算肿瘤抑制率;剩余裸鼠观察带瘤生存期;TUNEL法检测原位移植瘤细胞的凋亡指数;实时荧光定量PCR法和Western blot法检测凋亡相关基因Bcl-xL、Bax mR-NA和蛋白的表达。结果 5-Fu组、BL组、BM组、BH组的肿瘤抑制率分别为69.6%、45.6%、56.2%、58.5%,肿瘤体积与NS组比较明显缩小(P<0.01);BL组、BM组带瘤生存期与NS组相比较明显延长(P<0.05)。TUNEL结果显示蟾毒灵用药组的凋亡指数明显高于NS组(P<0.01);荧光定量PCR和Western结果显示蟾毒灵可抑制Bcl-xL基因表达,促进Bax基因表达(P<0.05)。结论蟾毒灵对裸鼠大肠癌原位移植瘤有显著的抗肿瘤作用,能够抑制Bcl-xL基因和促进Bax基因表达,诱导肿瘤细胞凋亡可能是蟾毒灵抗肿瘤的重要机制之一。

人U87-MG脑胶质瘤细胞裸鼠原位移植模型的建立

目的应用不同数量U87-MG细胞接种裸鼠脑内,建立裸鼠脑胶质瘤模型,观察其生长特性。方法采用立体定向技术,分别将3.0×10^(10)·L^(-1)、4.0×10^(10)·L^(-1)、5.0×10^(10)·L^(-1)浓度的人脑胶质瘤细胞U87-MG单细胞悬液,接种于18只♂裸鼠的右侧尾状核区,每只接种体积为5μL,另取6只裸鼠作为对照,接种同体积Hanks液。观察不同接种量实验鼠的生存状态、成瘤情况及脏器转移灶、带瘤生存期,测量肿瘤的最大径,计算肿瘤体积,将获取的全脑标本制作病理切片,行HE染色和免疫组化检查。结果各接种量的实验组成瘤率均为100%,未见颅外转移病灶,3组裸鼠的平均生存时间分别为(46.50±3.27)d、(38.50±3.28)d、(30.67±3.51)d;病理学检查符合人脑胶质瘤细胞的形态学特征和免疫表型;免疫组化GFAP和S-100蛋白呈阳性表达。结论立体定向脑内定量注射U87-MG细胞制备的人脑胶质瘤裸鼠原位移植模型成瘤率高、颅内生长稳定、颅外远隔部位转移率低,与人脑胶质瘤病理学及形态学特征相似,能为研究胶质瘤的发生、发病机制、生物学特性以及探寻有效的治疗措施提供可靠的动物模型。

优化胶粘贴法建立裸鼠胃癌原位种植模型

目的通过对两种胶粘贴法建立的胃癌原位种植动物模型的比较研究,为探讨胃癌的发病机制和实验治疗提供理想的动物模型。方法用OB胶和FS生物蛋白胶法分别建立胃癌原位种植动物模型,观察和比较两种方法所建立的模型肿瘤生长状况、转移情况和形态学变化。结果FS生物蛋白胶组未出现肿瘤大片坏死,腹水形成率为85.7%,幽门梗阻发生率为57.1%;而OB胶组肿瘤大片坏死发生率为100%,腹水形成率为14.3%,未出现幽门梗阻。FS生物蛋白胶组有三例出现了肺和脑转移。结论FS生物蛋白胶法建立的裸鼠胃癌原位种植动物模型能更好的模拟人胃癌患者的临床过程,为研究人胃癌转移机制和实验治疗提供理想的动物模型。

人体增生性疤痕移植于裸鼠动物模型的建立及稳定性观察

目的：将增生性疤痕组织块以不同的方法移植于裸鼠，建立一稳定的疤痕动物模型，为研究增生性疤痕提供实验基础。方法：将３ ｍｍ ×３ ｍｍ 增生性疤痕组织块分别移植于裸鼠皮下和体表创面，定期进行大体观察，组织学观察及胶原定量测定。结果：增生性疤痕组织块移植于裸鼠皮下后，移植物可成活８０～９０ ｄ，无排斥反应，并保留原增生性疤痕特性；移植物体积随着移植时间而逐渐增加。结论：增生性疤痕组织块移植于裸鼠皮下建立的增生性疤痕动物模型是一成功、稳定的动物模型，为疤痕的研究提供了可靠的基础。应用该动物模型时，作者建议把移植后３～１０ 周的动物作为增生性疤痕的标准动物模型。

LJH-OS人骨肉瘤裸鼠原位移植模型的建立及其生物学特性

用人成骨肉瘤细胞系LJH- OS的传代移植瘤组织作为移植材料,进行胫骨原位移植及皮下移植。结果发现胫骨原位移植的潜伏期较短,生长快。皮下移植瘤呈局限性膨胀性生长,有不完整的纤维包膜,未见肺转移,观察7 周无明显消瘦;而胫骨原位移植瘤浸润基层,无纤维包膜,且发生肺转移,7 周时有明显消瘦。原位移植的裸鼠血清ALP水平高于皮下移植者。说明裸鼠胫骨微环境较皮下组织更适于人骨肉瘤的浸润及转移表达,裸鼠胫骨原位移植模型的恶性生物学行为更接近临床骨肉瘤患者的实际情况,该原位移植模型的建立为骨肉瘤的研究提供了良好的实验模型。

应用蛋白质组学技术筛选鉴定锌指蛋白139调控的胃癌转移相关蛋白质

目的以胃癌SGC7901细胞原位移裸鼠模型为对象,通过荧光双向差异凝胶电泳(2D-DIGE)结合液质联用(LC-MS)质谱分析技术鉴定ZNF139调控的胃癌转移相关蛋白质。方法合成针对ZNF139的小干扰RNA(ZNF139-si RNA),以ZNF139-si RNA转染人胃癌细胞株SGC7901,G418筛选。以ZNF139-si RNA质粒转染的胃癌细胞、阴性质粒转染的胃癌细胞及普通胃癌细胞分别进行裸鼠胃癌原位移植。造模成功后取出原位移植瘤及腹腔转移淋巴结。荧光双向差异凝胶电泳(2D-DIGE)技术分别分离ZNF139-si RNA各组原位移植瘤及腹腔转移淋巴结蛋白质;选定差异点,胶内酶解后,液质联用(LC-MS)质谱分析技术鉴定蛋白质。蛋白印迹(Western blot)技术验证差异蛋白质的表达。结果转染ZNF139-si RNA质粒后SGC7901细胞中ZNF139的表达受到有效抑制。与阴性质粒组及空白组比较,阳性质粒组原位移植瘤生长更慢(P<0.05),且腹腔淋巴结转移率更低(P<0.05)。蛋白质组学结果发现在阳性质粒组原发灶中Fascin、hn RNPA2/B1表达下调,ANXA1表达上调;阳性质粒组转移淋巴结中ANXA5表达下调(P<0.05)。Western blot验证结果与蛋白质组学结果相符。结论 ZNF139可能通过调节Fascin、hn RNPA2/B1、ANXA1、ANXA5促进胃癌淋巴结转移。