人胃癌裸鼠原位移植模型的生物学行为研究

目的建立人胃癌裸鼠胃壁原位移植瘤模型，并与相应的皮下移植瘤作比较，以探讨宿主器官微环境对胃癌细胞浸润及转移等生物学行为的影响．方法将人胃癌细胞系ＭＧｃ８０３及其克隆株Ｃ１１癌细胞分别接种于裸鼠胃壁及背部皮下，比较原位和皮下移植瘤的成瘤率、生长率、生长方式及浸润、转移等生物学行为，以及体外回复培养后瘤细胞的增殖能力．结果胃壁原位及皮下移植瘤体内成瘤率、生长率及形态学上均无明显不同；其体外增殖能力也无显著性差异．但皮下移植瘤多呈局限性生长，无肝、脾、肾转移，其转移仅限于肺及个别局部淋巴结．胃壁原位移植瘤则浸润破坏胃壁各层组织结构，并直接蔓延到邻近各器官组织．其转移既有经血道至肝、肺、脾、肾等部位，也有经淋巴道至多数局部及远处淋巴结，其淋巴结的转移率较皮下移植瘤有显著增高（Ｐ＜００５）；且多伴有腹、盆腔内广泛种植性转移．结论裸鼠胃壁微环境较皮下组织更适合于人胃癌ＭＧｃ８０３及Ｃ１１移植瘤的浸润及转移的表达，该原位移植瘤模型的恶性生物学行为更接近临床胃癌患者的体内侵袭及转移的实际．     

人结肠鳞癌直肠鳞腺癌SCID鼠原位移植高转移模型的建立及生物学特性的研究

目的建立人结肠鳞癌直肠鳞腺癌SCID鼠原位移植高转移模型,为探讨理想的治疗结直肠癌肝转移和预防结直肠癌根治术后肝转移的方法提供实验工具.方法采用组织学完整的结直肠癌手术标本植入SCID鼠结直肠(粘膜层)壁内,观察原位移植的成瘤、移植瘤的侵袭和转移及其形态学特性(光镜、电镜、免疫组织化学).结果两株人结直肠癌SCID鼠均获原位移植成功.人直肠鳞癌SCID鼠原位移植模型HCS-HMN-1已传至19代,人直肠鳞腺癌SCID鼠原位移植模型HRSA-HMN-2已传至23代,共移植SCID鼠223只,其移植生长率和自发转移率及液氮冻存复苏成活率均为100%,移植瘤在结直肠内呈广泛原位侵袭性生长、淋巴结转移、肝转移和全腹腔癌病.并具有分泌CEA的功能.移植瘤细胞病理学和电镜观察、流式细胞仪DNA含量检测及染色体核型分析与瘤源人结直肠癌细胞完全一致.结论本研究所建立的两株人结直肠癌SCID鼠高转移模型完整模拟了人结直肠癌侵袭和转移的临床过程,为进一步研究结直肠癌肝转移治疗和预防结直肠癌根治术后肝转移的方法提供了理想的实验动物模型.     

恶性淋巴瘤的造血干细胞移植

恶性淋巴瘤（ＭＬ）是血液系统最常见的恶性肿瘤之一,其治疗效果与化疗剂量成正相关,剂量越大,治疗效果越好;但大剂量化、放疗的同时可增加药物的毒副反应,尤其是骨髓抑制作用,故限制了大剂量化、放疗的临床应用。近年来,采用造血干细胞移植（ＨＳＣＴ）治疗ＭＬ,...     

恶性淋巴瘤的造血干细胞移植治疗

对于常规化疗 5年生存率超过 5 0 %的预后较好、进展缓慢的淋巴瘤 ,目前不主张首先使用造血干细胞移植 (HSCT)治疗。1　 HSCT的适应证1.1　异基因造血干细胞移植 (Allo- HSCT)　尽管 Allo-HSCT具有移植物抗肿瘤效应 (GVT)、移植物中无肿瘤细胞污染、复发率低等优点 ,但其多被供     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

目的 探讨T细胞与B细胞起源的原发性结直肠恶性淋巴瘤(PCML)临床特征的差异.方法 回顾性分析1999年1月至2008年5月南昌大学第一附属医院经病理确诊的40例PCML患者的临床、内镜及病理资料.结果 40例PCML患者平均年龄为(41.0±11.2)岁,T淋巴细胞起源者占32.5%(13/40),B淋巴细胞起源者占67.5%(27/40).T淋巴细胞起源的PCML患者多见发热、盗汗、便血症状,易并发肠穿孔.内镜下T淋巴细胞起源的PCML患者多见多发或全结肠溃疡型病灶,B淋巴细胞起源者多见右半结肠隆起型单发病灶.B淋巴细胞起源的PCML确诊时临床分期较早,手术切除机会更多.结论 PCML内镜下肉眼观察诊断率较低,T淋巴细胞起源的PCML多为全结肠多发性溃疡型病灶,B淋巴细胞起源的PCML多为右半结肠隆起型单发病灶.B淋巴细胞起源的PCML确诊时临床分期较早.     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

Objective To investigate the difference of clinical features between primary colorectal malignant T cell lymphoma and primary colorectal malignant B cell lymphoma.Methods Forty cases diagnosed as primary eolorectal malignant lymphoma (PCML) between Jan 1999 and May 2008 were studied retrospectively.Results The average age of 40 patients with PCML was (41.0±11.2) years old.Twenty-seven cases (67.5%) were diagnosed as B-cell lymphoma.Thirteen cases (32.5%) were diagnosed as T-cell lymphoma.Patients with T-cell lymphoma in PCML had more symptoms such as fever,night sweat and hematochezia,and was easier to perforate than those with B-cell lymphoma.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under colonoscopy,but ulcerative type and multi-focus and fulbcolon were common in T-cell lymphoma.B-cell lymphoma had an earlier diagnosis,and more opportunities for surgery than T-cell lymphoma.Conclusions The misdiagnostic rate for PCML was high.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under eolonoscopy,but ulcerative type and multi-focus and full-colon were common in T-cell lymphoma.B-cell lymphoma could be diagnosed earlier.     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

Objective To investigate the difference of clinical features between primary colorectal malignant T cell lymphoma and primary colorectal malignant B cell lymphoma.Methods Forty cases diagnosed as primary eolorectal malignant lymphoma (PCML) between Jan 1999 and May 2008 were studied retrospectively.Results The average age of 40 patients with PCML was (41.0±11.2) years old.Twenty-seven cases (67.5%) were diagnosed as B-cell lymphoma.Thirteen cases (32.5%) were diagnosed as T-cell lymphoma.Patients with T-cell lymphoma in PCML had more symptoms such as fever,night sweat and hematochezia,and was easier to perforate than those with B-cell lymphoma.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under colonoscopy,but ulcerative type and multi-focus and fulbcolon were common in T-cell lymphoma.B-cell lymphoma had an earlier diagnosis,and more opportunities for surgery than T-cell lymphoma.Conclusions The misdiagnostic rate for PCML was high.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under eolonoscopy,but ulcerative type and multi-focus and full-colon were common in T-cell lymphoma.B-cell lymphoma could be diagnosed earlier.     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

Objective To investigate the difference of clinical features between primary colorectal malignant T cell lymphoma and primary colorectal malignant B cell lymphoma.Methods Forty cases diagnosed as primary eolorectal malignant lymphoma (PCML) between Jan 1999 and May 2008 were studied retrospectively.Results The average age of 40 patients with PCML was (41.0±11.2) years old.Twenty-seven cases (67.5%) were diagnosed as B-cell lymphoma.Thirteen cases (32.5%) were diagnosed as T-cell lymphoma.Patients with T-cell lymphoma in PCML had more symptoms such as fever,night sweat and hematochezia,and was easier to perforate than those with B-cell lymphoma.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under colonoscopy,but ulcerative type and multi-focus and fulbcolon were common in T-cell lymphoma.B-cell lymphoma had an earlier diagnosis,and more opportunities for surgery than T-cell lymphoma.Conclusions The misdiagnostic rate for PCML was high.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under eolonoscopy,but ulcerative type and multi-focus and full-colon were common in T-cell lymphoma.B-cell lymphoma could be diagnosed earlier.     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

Objective To investigate the difference of clinical features between primary colorectal malignant T cell lymphoma and primary colorectal malignant B cell lymphoma.Methods Forty cases diagnosed as primary eolorectal malignant lymphoma (PCML) between Jan 1999 and May 2008 were studied retrospectively.Results The average age of 40 patients with PCML was (41.0±11.2) years old.Twenty-seven cases (67.5%) were diagnosed as B-cell lymphoma.Thirteen cases (32.5%) were diagnosed as T-cell lymphoma.Patients with T-cell lymphoma in PCML had more symptoms such as fever,night sweat and hematochezia,and was easier to perforate than those with B-cell lymphoma.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under colonoscopy,but ulcerative type and multi-focus and fulbcolon were common in T-cell lymphoma.B-cell lymphoma had an earlier diagnosis,and more opportunities for surgery than T-cell lymphoma.Conclusions The misdiagnostic rate for PCML was high.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under eolonoscopy,but ulcerative type and multi-focus and full-colon were common in T-cell lymphoma.B-cell lymphoma could be diagnosed earlier.     

不同淋巴细胞起源的原发性结直肠恶性淋巴瘤临床特点差异分析

Objective To investigate the difference of clinical features between primary colorectal malignant T cell lymphoma and primary colorectal malignant B cell lymphoma.Methods Forty cases diagnosed as primary eolorectal malignant lymphoma (PCML) between Jan 1999 and May 2008 were studied retrospectively.Results The average age of 40 patients with PCML was (41.0±11.2) years old.Twenty-seven cases (67.5%) were diagnosed as B-cell lymphoma.Thirteen cases (32.5%) were diagnosed as T-cell lymphoma.Patients with T-cell lymphoma in PCML had more symptoms such as fever,night sweat and hematochezia,and was easier to perforate than those with B-cell lymphoma.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under colonoscopy,but ulcerative type and multi-focus and fulbcolon were common in T-cell lymphoma.B-cell lymphoma had an earlier diagnosis,and more opportunities for surgery than T-cell lymphoma.Conclusions The misdiagnostic rate for PCML was high.Protrude type and single-focus and right-colon type were common in B-cell lymphoma under eolonoscopy,but ulcerative type and multi-focus and full-colon were common in T-cell lymphoma.B-cell lymphoma could be diagnosed earlier.     

高转移MHCC97H细胞裸鼠原位移植瘤姑息性肝癌切除模型的建立

目的 模拟临床肝癌切除病例构建姑息性肝切除裸鼠模型,用于研究手术对残癌的影响并探索干预策略. 方法 采用肝左叶单瘤源接种技术,构建高转移潜能MHCC97H细胞裸鼠原位移植瘤模型,成功建模后14 d行姑息性肝切除.姑息术后14d,分析肿瘤相关基因差异表达;观察35 d,检测肝内、肺脏及腹腔转移;剩余裸鼠用于观察生存期.计量资料比较用t检验,生存分析用Kaplan-Meier法(Log Rank检验),P＜0.05为差异有统计学意义. 结果 原位移植瘤模型成功率100％,成功构建裸鼠姑息性肝切除模型,无手术相关死亡裸鼠.姑息组裸鼠肝内转移结节多于假手术组[(11.7±4.7)个对比(6.3±2.8)个,t=-2.412,P＜0.05],腹腔转移结节多于假手术组[(9.8±3.4)个对比(5.2±2.6)个,t=-2.641,P＜0.05],肺转移结节也多于假手术组[(14.3±4.7)个对比(8.7±4.7)个,t=-2.348,P＜0.05].应用生物信息学技术进一步分析发现,肿瘤转移抑制蛋白1、肿瘤坏死因子β1、Smad2、白细胞介素1β及基质金属蛋白酶7基因在姑息组残癌基因功能网络中处于核心位置.姑息组裸鼠生存期为(60.8±2.7)d,假手术组裸鼠生存期为(51.3±1.4)d,差异有统计学意义(x2=12.850,P＜0.01).结论 单瘤源姑息性肝切除模型能有效模拟临床肝切除病例,为术后残癌生物学特性的研究及干预提供一个新的平台.     

Liver metastasis models of human colorectal carcinoma established in nude mice by orthotopic transplantation and their biologic characteristic

AIM To establish a liver metastasis model of human colorectal carcinoma in nude mice.METHODS Orthotopic transplantation of histologically intact colorectal tissues from patients into colorectal mucosa of nude mice. Tumorgenicity, invasion, metastasis and morphological characteristics of the transplanted tumors were studied by light microscopy, electron microscopy and immunohistochemistry.RESULTS Liver metastasis models of human colon carcinoma (HCA-HMN-1) and human rectal carcinoma (HRA-HMN-2) were established after screening from 34 colorectal carcinomas. They had been passaged in vivo for 18 and 21 generations respectively. There were lymphatic, hemotogenous and implanting metastasesis. CEA secretion was maintained after transplantation. The primary and liver metastatic tumors were similar to the original human carcinoma in histopathological and ultrastructural features, DNA content and chromosomal karyotype.CONCLUSION The liver metastasis models provide useful tools for the study of mechanism of metastasis and its treatment of human colorectal cancer.INTRUDUCTIONSome models of nude mice that fresh human colorectal carcinoma tissue or cells were successfully transplanted subcuteneously have been reported at home and abroad[1,2]. But until now there has been no report on a liver metastasis model of human colorectal carcinoma established by orthotopic transplantation in nude mice in China. Based on our previous models of human liver and pancreas carcinoma by orthotopic transplantation[3,4], we established liver metastasis models of colon and rectum carcinoma with a spontaneous metastasis rate of 100%.     

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.     

三羟基异黄酮诱导人胃癌原代细胞裸鼠移植瘤凋亡的研究

目的探讨三羟基异黄酮诱导人胃癌原代细胞裸鼠移植瘤凋亡的机制。方法建立人胃癌原代细胞裸鼠移植瘤模型并测定移植瘤的生长曲线;25只Balb/c裸鼠接种胃癌原代细胞悬液生成移植瘤后均分为五组,不同剂量的三羟基异黄酮(0.5、1.0和1.5mg/kg)、生理盐水和DMSO进行瘤旁注射,透射电镜和TUNEL法检测肿瘤组织凋亡情况,免疫组化法和RT-PCR法检测肿瘤组织凋亡相关基因bcl-2和bax的表达情况。结果三羟基异黄酮对胃癌原代细胞移植瘤有明显的抑制作用;透射电镜发现三羟基异黄酮导致移植瘤内大量细胞发生凋亡;TUNEL染色法发现三羟基异黄酮注射组瘤细胞的凋亡指数(28.7%±1.1%、33.4%±1.4%和37.1%±1.0%)明显高于生理盐水和DMSO对照组(P<0.05),而两个对照组(11.8%±0.4%和11.7%±0.4%)之间差异无统计学意义。免疫组化法发现三羟基异黄酮注射组的Bcl-2蛋白阳性细胞率(11.8%±0.9%、5.7%±0.8%和4.0%±0.8%)明显低于两个对照组(P<0.05),而两个对照组(19.5%±0.6%和17.6%±1.4%)之间差异无统计学意义;而Bax蛋白阳性细胞率(20.0%±1.2%、24.7%±0.9%和29.3%±1.6%)明显高于两个对照组(P<0.05),而两个对照组(10.3%±0.4%和10.9%±0.9%)间差异无统计学意义;RT-PCR法发现三羟基异黄酮注射组的bcl-2mRNA条带强度随剂量增大而递减,并明显低于对照组(P<0.05);而baxmRNA条带强度递增,并明显低于对照组(P<0.05),而两对照组间差异无统计学意义(P>0.05)。结论三羟基异黄酮对胃癌原代细胞移植瘤有明显的抑制作用,通过下调bcl-2和上调bax的表达而诱导胃癌裸鼠移植瘤细胞发生凋亡,是其抗胃癌作用的机制之一。     

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 apoptosisregulated 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 morphological changes 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     

两种细胞制作的人肝癌裸鼠模型转移性状的比较

目的 比较两种人肝癌细胞株(HCCLM3和SMMC-7721细胞)构建的人肝癌裸鼠模型的转移潜能.方法 实验分为两组:应用HCCLM3细胞构建的人肝癌裸鼠模型组为A组,SMMC-7721细胞构建的模型组为B组.比较A组和B组的原发灶的肿瘤大小和病理检查结果.比较各组的肝内转移率、转移数目,累及的肝叶数以及转移肿瘤体积的大小.所有数据由SPSS16.0软件分析,计量资料采用t检验,率的比较采用Fisher确切概率法,计数资料采用秩和检验.结果 A组的肝内转移率为100%(8/8)、肝内原发肿瘤体积为(6954±1945)mm~3,B组的肝内转移率为62.5%(5/8)、肝内原发肿瘤体积为(6034±2035)mm~3,两组肝内转移率和原发灶肿瘤体积大小比较,差异无统计学意义.肝内转移的数目、累及肝叶数、转移肿瘤的体积大小A组分别为4.5个(中位数)、3个和975 mm~3(中位数),B组分别为1个(中位数)、1个和274 mm~3(中位数),两组比较,Z值分别为-2.818、-2.289和-1.975,P值均<0.05,差异均有统计学意义.两组在肺转移率(P=0.001)和累及其他脏器个数(P=0.041)的比较,差异均具有统计学意义.结论 HCCLM3细胞制作的人肝癌裸鼠转移模型具有高转移潜能,可应用于人肝癌的生物性状的研究.     

小鼠结肠癌完整组织块原位种植及肝转移模型的建立方法

[目的]研究快速建立模拟临床过程的小鼠结肠癌原位移植瘤及肝转移模型方法。[方法]采用小鼠结肠癌细胞CT26接种于裸鼠皮下,形成稳定传代的皮下种植瘤。再取该肿瘤组织块,用原位医用胶黏合法种植于BALB/c小鼠盲肠壁,建立模拟于临床的结肠癌肝转移模型,定时观察小鼠全身情况,分不同时期处死,观察其生长及转移特性。[结果]原位种植成瘤率100%,术后3周肝转移率100%,4周后动物消瘦,萎靡,行动迟缓,反应迟钝,部分可见腹水形成,濒临死亡。中位生存期29d。[结论]小鼠结肠癌完整组织块通过"生物胶黏贴法"原位种植BALB/c小鼠盲肠壁,速度快,方法简便,能较好的重现结肠癌临床转移的过程,为人类结肠癌生长、肝转移机制及干预肝转移治疗的研究提供一种较理想的动物模型。     

Establishment of an orthotopic lung cancer model in nude mice and its evaluation by spiral CT

Objective

To establish a simple and highly efficient orthotopic animal model of lung cancer cell line A549 and evaluate the growth pattern of intrathoracic tumors by spiral CT.

Methods

A549 cells (5×10⁶ mL^-1) were suspended and inoculated into the right lung of BALB/c nude mice via intrathoracic injection. Nude mice were scanned three times each week by spiral CT after inoculation of lung cancer cell line A549. The survival time and body weight of nude mice as well as tumor invasion and metastasis were examined. Tissue was collected for subsequent histological assay after autopsia of mice.

Results

The tumor-forming rate of the orthotopic lung cancer model was 90%. The median survival time was 30.7 (range, 20-41) days. The incidence of tumor metastasis was 100%. The mean tumor diameter and the average CT value gradually increased in a time-dependent manner.

Conclusions

The method of establishing the orthotopic lung cancer model through transplanting A549 cells into the lung of nude mice is simple and highly successful. Spiral CT can be used to evaluate intrathoracic tumor growth in nude mice vividly and dynamically.Key words : Lung cancer, orthotopic lung cancer model, spiral CT, A549