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.     

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．     

下调CD44基因表达对人胃癌SGC7901细胞裸鼠移植瘤的抑制作用及其机制

目的探讨靶向CD44的短发夹RNA(shRNA)对人胃癌裸鼠移植瘤生长抑制作用及其机制。方法利用CD44 shRNA细胞及对照细胞株建立裸鼠原位移植瘤及皮下种植瘤模型,监测肿瘤生长变化,采用RT-PCR及免疫组织化学方法检测瘤体内CD44表达的变化,并进一步检测上皮-间质转化(EMT)相关因子表达的变化。结果成功构建了CD44 shRNA转染荷瘤皮下及原位裸鼠模型。与对照shRNA组、空白对照组分别比较,CD44 shRNA组荷瘤裸鼠肿瘤生长速度减慢,皮下种植瘤及原位移植瘤质量减轻,差异均有统计学意义(P均0.01)。RT-PCR结果发现,CD44 shRNA组CD44mRNA表达在皮下种植瘤及原位移植瘤均显著下调,差异均有统计学意义(P均0.01)。CD44 shRNA组细胞EMT相关基因E-cadherin表达增加,N-cadherin、Vimentin表达降低。结论 CD44 shRNA可以有效抑制人胃癌SGC7901细胞裸鼠移植瘤生长,并下调CD44的表达水平,这可能与CD44基因参与EMT相关。     

三种来源细胞因子诱导的杀伤细胞对人食管癌裸鼠原位移植肿瘤的转移抑制作用

目的探讨三种来源的细胞因子诱导的杀伤(CIK)细胞对人食管癌裸鼠原位移植肿瘤的转移抑制作用。方法建立人食管癌裸鼠原位移植模型。取32只荷瘤裸鼠平分为四组:食管癌患者自体外周血CIK细胞组、健康人外周血CIK细胞组、脐血CIK细胞组及生理盐水对照组。给予20 d对应的注射治疗,停药72 h后处死裸鼠,称量各组原位移植瘤瘤重,计算抑瘤率;观察肿瘤转移情况;免疫组织化学法检测基质金属蛋白酶-9(MMP-9)的表达情况。结果与对照组相比,CIK细胞治疗组的肿瘤生长及转移明显受到抑制(P<0.05)。且CIK细胞可抑制肿瘤组织中MMP-9的表达。结论三种来源CIK细胞对人食管癌裸鼠原位移植肿瘤的生长及转移有明显抑制作用,且脐血来源的CIK细胞抗瘤作用最强,其抗转移作用可能与肿瘤组织中MMP-9的表达下降有关。     

不同转移潜能食管癌细胞株裸鼠模型的建立

背景：食管癌的转移率较高,是导致患者死亡的主要原因,但其机制仍不完全清楚。目的：建立具有不同转移潜能的高侵袭能力食管癌细胞株亚系裸鼠模型。方法：应用Transwell侵袭小室从食管癌细胞株Eca-109中筛选出高侵袭能力的食管癌细胞株亚系。观察母系和亚系细胞形态,MTT法检测细胞增殖能力的变化,划痕法检测细胞迁移能力。将食管癌Eca-109细胞及其亚系Eca-109 T4细胞分别皮下注射于裸鼠体内诱导移植瘤模型,观察成瘤率、成瘤时间、肿瘤生长情况。结果：成功从食管癌Eca-109细胞株中筛选出高侵袭能力的食管癌细胞株亚系Eca-109 T4,两者细胞形态无明显差异。与Eca-109细胞相比,Eca-109 T4细胞增殖能力和划痕愈合能力均明显增强。Eca-109组和Eca-109 T4组裸鼠成瘤率均为100%,与Eca-109组相比,Eca-109 T4组裸鼠成瘤时间更早且瘤体生长更快。结论：成功建立了不同转移潜能的高侵袭能力食管癌细胞株亚系裸鼠成瘤模型,为食管癌转移的进一步研究提供了理想模型。     

肺原发性霍奇金淋巴瘤临床病理分析

目的 探讨肺原发性霍奇金淋巴瘤的诊断与鉴别诊断.方法 回顾性分析3例肺霍奇金淋巴瘤病例并复习相关文献.结果 3例患者均为男性,以咳嗽症状为主,肿瘤较大,平均直径为5 cm,可伴有肺门及纵隔淋巴结肿大,镜下均可见典型的RS细胞,背景细胞内有较明显嗜酸粒细胞浸润,并见干尸细胞,免疫组化CD15和CD30阳性.化疗、骨髓移植可使病情缓解,长期生存.结论 肺原发性霍奇金淋巴瘤病理学及免疫表型具有一定的特征,恰当的治疗预后良好.     

抗转移多肽对人胃癌腹膜高转移细胞侵袭转移的抑制作用

目的研究多肽PⅢ对人胃癌腹膜高转移细胞系GC9811-P腹膜转移的作用。方法利用体外细胞基质粘附和体外细胞侵袭实验,分别检测多肽PⅢ对GC9811-P细胞粘附及侵袭能力。体内实验采用胃浆膜下裸鼠原位接种转移模型,观察多肽PⅢ对胃癌细胞GC9811-P腹膜转移能力的影响。裸鼠分多肽PⅢ治疗组、0．9%氯化钠溶液对照组各12只,荷瘤鼠衰竭处死后观察腹膜转移率、转移灶数目及原发瘤质量。结果40μg多肽PⅢ孵育2h时粘附抑制率达86．3%,多肽PⅢ与GC9811-P细胞共同孵育对层粘连蛋白粘附的抑制作用呈时间依赖性。多肽PⅢ与GC9811-P细胞共孵育48h后侵袭抑制率达81．4%。将肿瘤细胞原位接种于裸鼠后给予多肽PⅢ治疗,与对照组比较腹膜转移灶的数目分别为(3．2±6．5)个和(26．3±5．2)个,腹膜转移率明显下降(P＜0．01)；而原发瘤质量分别为(1．9±1．2)g和(2．1±1．0)g,两者间差异无统计学意义(P＞0．05)。结论多肽PⅢ明显抑制人胃癌腹膜高转移细胞系GC9811-P粘附、侵袭和腹膜转移作用。     

间变性大细胞淋巴瘤的临床细胞病理学分析

目的:探讨间变性大细胞淋巴瘤(anaplastic large cell lymphoma,ALCL)的临床细胞病理学特点、免疫表型及鉴别诊断,提高对ALCL的认识。方法:分析经组织病理学证实的13例ALCL的临床表现、细胞病理学形态及免疫表型,并结合相关文献进行讨论。结果:13例ALCL患者中,ALK阳性11例,ALK阴性2例;全部表达CD30,部分表达EMA、CD3、CD45RO、CD43,不表达CD3、CD56、CD20、CD79a、PCK、CD117;Ki67LI 60%~100%。结论:ALCL是一种少见的非霍奇金淋巴瘤,依据其细胞病理学形态特征,细胞病理学可以做出正确的诊断,但应注意与霍奇金淋巴瘤、弥漫大B细胞淋巴瘤、恶性黑色素瘤、未分化癌等进行鉴别;当鉴别诊断有困难时,应行活检及免疫组织化学染色以确诊。     

CD133+肺癌细胞的干细胞特性研究

目的 收获CD133+肺腺癌细胞并评估其肿瘤干细胞特性.方法 从13例新鲜肺腺癌组织中收获CD133+和CD133-肺腺癌细胞,并进行Transwell侵袭实验、裸鼠成瘤实验等对比研究,观察CD133+和CD133-肺腺癌细胞的差异.结果 13例中有10例发现CD133表达,其阳性表达率最高为13.12%,CD133+和CD133-细胞在侵袭性、致瘤能力上有明显区别.结论 CD133在人肺腺癌组织细胞中广泛表达,CD133+较CD133-人肺腺癌细胞具有更强的侵袭性和致瘤能力.     

原发性肝脏淋巴瘤的临床病理特点及其外科治疗

目的:探讨原发性肝恶性淋巴瘤(primary liver lymphoma,PLL)的临床病理特点及诊治方法.方法:对我院1975-2008年收治的经病理确诊的7创原发性肝恶性淋巴瘤的病因、临床表现、形态学光镜、电镜、免疫组织化学、血清学甲胎蛋白(AFP)、乙型肝炎(HBsAg)、乳酸脱氢酶(LDH)和治疗结果进行系统分析.结果:本组7例患者,4例均上腹痛发热,3例有淋巴瘤B症状,发热、盗汗、体质量减轻.有5例合并慢性肝炎或肝硬化.组织病理学5例为B细胞表型.免疫组织化学显示CD20、CD79α阳性,电镜示瘤细胞间毛细血管、胞质细胞器发达.2例为T细胞表型,CD3、CD45RO阳性,电镜下瘤细胞质稀疏,细胞器不发达.血清学检查5例AFP和CEA阴性.5例HBsAg阳性,2例HBsAg阴性.LDH 675.54 U/L-1246.5 U/L.4例术后采用化疗者生存时间平均为9.5 mo.3例术后采用生物化疗者生存时间为23.0 mo.结论:PLL与HBV病毒感染有关:伴有B症状,常见肝内占位性病变和LDH增高:首选手术切除联合生物化疗的治疗模式:能够延长患者的生存时间.     

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%.     

CD44V6对实验性大肠癌肝转移影响的实验研究

目的:研究 CD44V6在大肠癌肝转移中的作用,探索一种有效的抗肿瘤转移方法。方法:应用大肠癌肝转移模型来研究 CD44v6单抗对大肠癌 HT29、Lovo 细胞裸鼠实验性肝转移的影响,分析 CD44V6单抗在抗大肠癌肝转移中的作用。结果:CD44V6单抗能有效阻断裸鼠实验性大肠癌肝转移的发生。     

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

ＩＮＴＲＵＤＵＣＴＩＯＮＳｏｍｅｍｏｄｅｌｓｏｆｎｕｄｅｍｉｃｅｔｈａｔｆｒｅｓｈｈｕｍａｎｃｏｌｏｒｅｃｔａｌｃａｒｃｉｎｏｍａｔｉｓｕｅｏｒｃｅｌｓｗｅｒｅｓｕｃｃｅｓｆｕｌｙｔｒａｎｓｐｌａｎｔｅｄｓｕｂｃｕｔｅｎｅｏｕｓｌｙｈａｖｅｂｅｅｎｒ...     

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.     

Study of orthotopic transplantation model of human gastrointestinal cancerand detection of micrometastases

AIM To establish a relevant animal model of human gastrointestinal cancer, which can be used forrepetitive investigations and may improve our understanding of carcinogenesis and cancer metastasis.METHODS Intact tissue of human colorectal and pancreatic cancers was transplanted in nude mice. Thebiological characteristics of the original and corresponding transplanted tumors were investigated by HEstaining, PAS staining and immunostaining. The metastases in livers and lungs of the nude mice wereinvestigated by immunostaining with biotinylated mab KL-1 and by RT-PCR using CK20 specific primers.RESULTS Nine of 16 surgical specimens grew in the nude mice subcutaneously and/or orthotopically (4 of6 colorectal and 5 of 10 pancreatic cancer). Tumor cell content of the specimens and freezing of tissuespecimens are important factors influencing the growth of transplanted tumor. In the group of fresh tumortissues with greater than 50% tumor cell content, transplantation rate was 100% (3 cases of pancreatic cancerand 3 cases of colorectal cancer). The orthotopically transplanted tumors resembled the original tumormorphologically and biologically, including TAA expression such as CEA by immunohistochemistry, andCEA level in the serum of mice. Ki-67 labeling index and the expression of TAA especially K-ras, 17-1A and RA-96, were associated with the potential of tumor growth in nude mice. Micrometastases in the lungs andlivers of tumor bearing mice could be detected by immunostaining with biotinylated mab KL-1 and CK20-sepcific RT-PCR.CONCLUSION An orthotopic transplantation model for human colon and pancreatic cancer in nude micehas been established. The sensitive detection methods with CK-immunohistochemistry and CK20-RT-PCRwere also established to study xenotransplanted human cancer and its metastatic cancer cells in the liver andlung of nude mice. This study may be helpful in understanding the mechanism of cancer metastasis and indeveloping new diagnostic methods and therapeutic strategies for metastases.Acknowledgement The authors thank Dr. J. Luettges, Department of Pathology; Kiel University, for investigating thepathological characterics of the specimens; Dr. N. Zawazawa, Institute of Immunology, Kiel University, for the quantitativemeasurement of serum of CEA.     

Orthotopic transplantation model of human gastrointestinal cancer and detection of micrometastases

AIM To establish a relevant animal model ofhuman gastrointestinal cancer,which can beused for repetitive investigations,so as toimprove our understanding and management ofcarcinogenesis and cancer metastasis.METHODS Intact tissues of human colorectaland pancreatic cancers were transplanted innude mice.The biological characteristics of theoriginal and the corresponding transplantedtumors were investigated by HE staining,PASstaining and immunostaining.The metastases inthe livers and lungs of nude mice wereinvestigated by immunostaining withbiotinylated mab KL-1 and by RT-PCR using CK20specific primers.RESULTS There were totally 9 of 16 surgicalspecimens growing in nude mice subcutaneouslyand/or orthotopically(4 of 6 colorectal and 5 of10 pancreatic cancer).Tumor cell content of thespecimens and freezing of tissue specimens areimportant factors influencing the growth oftransplanted tumor.In the group of fresh tumortissues with greater than 50% tumor cell content,the success rate of the transplantationwas 100%(3 cases of pancreatic cancer and 3cases of colorectal cancer).The orthotopicallytransplanted tumors resemble the original tumormorphologically and biologically,including TAAexpression such as CEA byimmunohistochemistry,and CEA level in theserum of mice.Ki-67 labeling index and theexpression of TAA especially K-ras,17-1A andRA-96,are associated with the potential of tumorgrowth in nude mice.Micrometastases in thelungs and livers of tumor bearing mice can bedetected by immunostaining with biotinylatedmab KL-1 and CK20-specific RT-PCR.CONCLUSION An orthotopic transplantationmodel for human colon and pancreatic cancer innude mice has been set up.We have alsoestablished sensitive detection methods withCK-immunohistochemistry and CK20-RT-PCR tostudy xenotransplanted human cancer and itsmetastatic cancer cells in the liver and lung ofnude mice.This study may be helpful inunderstanding the mechanism of cancermetastasis and in developing new diagnosticmethods and therapeutic strategies formetastases including micrometastases.     

Metastatic models of human liver cancer in nude mice orthotopically constructed by using histologically intact patient specimens

In this study of orthotopic implantation of histologically intact surgical specimens, the authors constructed metastatic models of human hepatocellular carcinoma (HCC) in nude mice. Histologically intact human liver cancer specimens, derived from patients, were implanted directly into the liver of nude mice, and their orthotopic growth and metastases were observed. The transplantability and metastatic rate of two specimen groups (primary and metastatic lesions) were analysed. -Fetoprotein (AFP) was also determined in transplanted tumours by an immunohistochemical method. Orthotopic growth was observed in 14 of 30 transplanted specimens and formation of metastases in 7 cases, which exhibited the variety of clinical behaviours seen in patients with HCC. These behaviours included local growth, regional invasion, spontaneous intrahepatic, lymph node and lung metastasis and peritoneal seeding. In two groups the growth rate of metastatic lesions following implantation was clearly higher than that of primary tumours. Chromosome analysis from locally growing tumours confirmed their morphologically human origin. An immunohistochemical study showed that implanted tumours originating from AFP-positive specimens maintained AFP expression. These results indicated that the animal models should prove valuable for developing new treatment modalities and studying the mechanism of metastasis of human HCC.Abbreviation HCC hepatocellular carcinoma - AFP -fetoprotein     

Nude mice model of human hepatocellular carcinoma via orthotopic implantation of histologically intact tissue

AIM: To establish a nude mice model of human hepatocellular carcinoma (HCC) via orthotopic implantation of histologically intact tissue, in order to study biologic features of HCC in vivo and to direct clinical treatment respectively.METHODS: Histologically intact fresh specimens of HCC were orthotopically implanted in nude mice (BALB/c, nu/nu). Survival rate and growth curve were investigated with Bultrasound. Morphological characteristics of pathology and spontaneous metastatic rates were detected with microscopy. Expression of multidrug resistance genes studied with immunohistochemical method and RT-PCR, and other biologic features of implanted tumor were observed and compared with human HCC specimens. RESULTS: Out of the specimens from two patients with HCC, only one specimen survived in nude mice. The orthotopic implantation tumor survival rate, spontaneous intrahepatic metastatic rate, pulmonary metastatic rate and bone metastases rate were 100%, 75.0%, 37.5% and 37.5% respectively in the first passage. AFP was kept on secreting and increasing with the size of the tumor. The morphological characteristics and biologic features were similar to the donor‘s, the protein and mRNA of MDR1 and LRP were expressed in tumors of the model and the donor, and there was no significant difference between them (P&gt;0.05). CONCLUTION: The model of nude mice with orthotopic implantation of histologically intact HCC tissue is an ideal model to study biologic features of HCC in vivo and to direct clinical treatment.