人类胃癌裸鼠淋巴结转移模型的建立

 目的　研究胃癌淋巴结转移动物模型的建立方法。方法　人类胃癌低分化细胞系SGC-7901体外培养、传代并扩增后，收集细胞行皮下种植成瘤，鼠间传代至第6代，以皮下肿瘤组织块原位种植于裸鼠胃壁建立动物模型。种植后第9周处死裸鼠，观察原位种植瘤生长、淋巴结转移及其他脏器转移情况，测定荷瘤裸鼠血清癌胚抗原（CEA）值。结果　原位移植瘤种植成功率100 ％，胃周淋巴结转移率93.3 ％，移植瘤可发生局部浸润及远处脏器转移，荷瘤裸鼠CEA值明显高于正常裸鼠（P＜0.01）。结论　应用SGC-7901细胞系可成功建立胃癌的淋巴结转移动物模型。     

人卵巢上皮癌裸鼠皮下移植瘤模型的建立及生物学性状的鉴定

目的　建立人卵巢上皮性癌裸鼠皮下移植瘤模型 ,为人卵巢癌的诊断及治疗诸方面提供理想的在体研究工具。方法体外培养人卵巢上皮癌细胞株SKOV 3 ,采用皮下注射方式接种至裸小鼠 ,待肿瘤直径达 0 .8cm～ 1.0cm时 ,将其切下剪成直径为1mm～ 2mm小块 ,再接种至另一裸鼠皮下进行传代。病理组织学检查或透射电镜观察肿瘤 ,并观察染色体特征 ,检测裸鼠血清和肿瘤组织中肿瘤标志物CA12 5、CEA、AFP的浓度。结果　成功地建立了人卵巢上皮性癌细胞株SKOV 3裸小鼠皮下移植瘤模型 ,并传至第二代 ,原代接种成功率为 69.2 % (2 7/ 3 9) ,鼠间传代移植成功率为 10 0 .0 % ,具备人肿瘤生物学特点 ,但未观察到转移行为。裸鼠血清CA 12 5浓度低 ,但在肿瘤冻融产物中较高。结论　建立的人卵巢癌裸鼠皮下移植瘤模型具有人卵巢癌特征 ,可用于研究卵巢癌的生物学特性 ,为卵巢癌临床诊断及治疗研究提供有用工具。     

裸鼠人胃腺癌SGC-7901原位移植模型的构建及其生物学特性

目的：建立裸鼠人胃癌原位移植模型。方法：以反复接种传代于裸鼠皮下的SGC-7901人胃癌细胞株建立的移植瘤组织块为材料，将其原位移植于裸鼠胃壁，观察移植肿瘤的生长情况、移植成功率和自发转移的发生率。结果：原位移植成功率及局部浸润率为8／8，局部淋巴结转移率为8／8，肺转移发生率为5／8，肝转移发生率为7／8，腹膜转移发生率为7／8。结论：裸鼠人胃癌原位移植模型的生物学行为与人胃癌自然生长和转移过程相似，可作为肿瘤转移机制及其抗转移治疗实验研究的理想模型。     

人卵巢癌裸鼠移植瘤和腹水瘤模型的建立及形态学观察

目的 建立人卵巢癌裸鼠移植模型,为研究人卵巢癌发病机制、实验治疗提供工具。方法 采用组织学完整 的人卵巢癌实体瘤和腹水瘤标本植入裸鼠腹腔和皮下,观察成瘤生长和转移以及形态学特征（光镜、电和染色体）。结果 10例卵巢癌标本2例实体瘤、1例腹水瘤移植成功,实体瘤传至第六代,腹水瘤传至第十八代,传代移植成功率100％,移植瘤病理学和电镜观察、染色体核型分析、肿瘤标记物检查均与人体原发肿瘤一致。结论 本研究建立的人卵巢癌实体瘤和腹水瘤裸鼠移植模型与临床患者相似,为进一步研究人体卵巢癌发病和治疗提供了理想的实验动物模型。     

人类胃癌裸鼠原位移植模型的建立

 目的：探讨建立人胃癌裸鼠原位移植转移模型简便易行的方法，比较三种方法的成瘤率。方法：用人胃癌细胞SGC7901悬液接种于裸鼠皮下，形成稳定传代的皮下移植瘤，连续传八代。15只裸鼠随机分为三组，以第八代裸鼠皮下移植瘤组织为移植材料，分别用困扎法、医用OB胶法以及两种联合的方法把皮下肿瘤组织移植裸鼠胃浆膜下形成原位移植肿瘤。结果：6～7周后，处死全部裸鼠，捆扎组、医用OB胶组和联合组成瘤率分别为4／5、3／5和5／5，捆扎联合医用OB组成瘤率最高。结论：应用缝线困扎联合OB胶建立人胃癌裸鼠原位移植模型是一种简便易行的方法，值得推广和应用。     

人骨肉瘤原位移植模型的建立及生物学特征

目的 用人骨肉瘤细胞系HOS-98建立人骨肉瘤裸鼠胫骨原位移植模型,以探讨宿主器官微环境对人骨肉瘤细胞侵袭及转移等生物学行为的影响。方法 将人骨肉瘤细胞系HOS-98接种于裸鼠皮下,形成移植瘤,用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。分别于移植后4周和8周处死小鼠,进行病理形态学检查,并对两种方法在成瘤率、生长方式及侵袭、转移等生物学行为比较。结果 两种移植方式在成瘤率及形态学上无明显不同,胫骨原位移植的潜伏期较短,并且生长快于皮下移植方式。皮下移植瘤呈局限性膨胀生长,有不完整的纤维包膜,瘤内类骨基质较少见,未见肺转移,观察8周时无明显消瘦;而胫骨原位移植瘤侵袭周围组织,可见发生肺转移,8周明显消瘦。原位移植的裸鼠血清ALP水平高于皮下移植者。原位移植的X线检查有明显的类似于人的骨性反应。结论 用人骨肿瘤细胞系HOS-98皮下接种的移植瘤作为移植材料是建立肿瘤异位移植的可行途径,裸鼠胫骨微环境较皮下组织更适合于人骨肉瘤的侵袭及转移表达,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体内侵袭及转移实际,该原位移植模型为今后的实验研究提供了更加接近患者实际的实验模型。     

人胃癌组织块裸鼠原位移植/转移模型的建立

 目的　用肿瘤组织块原位移植 ,建立人胃癌裸小鼠原位移植 /转移模型。方法　以人胃低分化腺癌细胞系接种于裸小鼠皮下 ,形成稳定传代的皮下移植瘤 ,再取该肿瘤组织块原位移植于裸鼠胃壁 ,观察移植肿瘤的生长状况、移植成功率和自发转移的发生率。结果　原位移植成功率 (成瘤率 )为 1 0 0 %、局部淋巴结转移率 1 0 0 %、远处淋巴结转移率 90 %、肝转移发生率为 75%。荷瘤鼠的中位生存期为 1 4周 ,晚期出现消瘦和全身衰竭。结论　该裸小鼠原位移植 /转移模型的生物学行为与人胃癌自然生长和转移过程相似 ,可作为一种有价值的工具用于胃癌转移机理和抗转移实验治疗的研究。     

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,已传至第26代.移植成功率达100%,平均裸鼠带瘤存活中位数为102天.肿瘤倍增时间为7.17(SD=±1.02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代.流式细胞仪及显微分光光度计检测移植瘤,提示肿瘤为多倍体,各代移植瘤的DNA指数与第一代肿瘤基本一致(DI1.05~1.40).经初步应用研究,用传代移植瘤组织提取抗原免疫BALB/C小鼠,将免疫小鼠的脾细胞与小鼠缺欠型骨髓瘤系进行融合,制备了一株抗卵巢癌单克隆抗体,经免疫组化染色(ABC法)可显示有与人卵巢癌阳性反应.说明裸鼠人体卵巢癌模型的建立,为深入进行该类肿瘤诊断及治疗研究提供了有效途径.     

裸鼠人胃腺癌SGC-7901原位移植模型的构建及其生物学特性

目的 :建立裸鼠人胃癌原位移植模型。方法 :以反复接种传代于裸鼠皮下的SGC 790 1人胃癌细胞株建立的移植瘤组织块为材料 ,将其原位移植于裸鼠胃壁 ,观察移植肿瘤的生长情况、移植成功率和自发转移的发生率。结果 :原位移植成功率及局部浸润率为 8 8,局部淋巴结转移率为 8 8,肺转移发生率为 5 8,肝转移发生率为 7 8,腹膜转移发生率为 7 8。结论 :裸鼠人胃癌原位移植模型的生物学行为与人胃癌自然生长和转移过程相似 ,可作为肿瘤转移机制及其抗转移治疗实验研究的理想模型     

4种人肾细胞癌原位移植裸鼠模型建立方法的比较

背景与目的:肾细胞癌是最常见的肾脏恶性肿瘤,起病隐匿,恶性程度高.研究旨在建立成功率高、稳定的人肾细胞癌原位动物造模方法.方法:采用人肾细胞癌细胞(786-0、ACHN)进行细胞悬液原位注射法、皮下成瘤后引瘤原代细胞悬液原位注射法、皮下成瘤后引瘤组织块原位移植肾包膜下法及皮下成瘤后引瘤组织块原位移植肾周筋膜内法建立原位移植裸鼠模型,采用PET/CT、H-E染色、免疫组织化学染色及血清生化检测等手段观察成瘤率及肿瘤生长情况,评估4种方法的建模效果.结果:人肾细胞癌皮下移植瘤裸鼠模型,ACHN组成瘤率(90%)高于786-0组(30%);人肾细胞癌原位移植裸鼠模型,786-0原位注射组、ACHN原位注射组、ACHN皮下移植瘤引瘤后原代细胞原位注射组、ACHN组织块肾包膜下包埋组和ACHN组织块肾周筋膜内包埋组的成瘤率分别为33%、80%、90%、100%和20%,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤率最高,影像学及病理组织学检查结果显示符合低分化肾细胞癌.结论:成功建立4种人肾细胞癌原位移植裸鼠模型,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤效果最佳,为肾细胞癌发病机制及靶向治疗的进一步研究提供理想的动物模型.     

卵巢癌高频转移细胞模型中nm23-H1基因表达的相关性研究

目的 筛选高频转移卵巢恶性肿瘤细胞，研究不同转移潜能的细胞和nm23的相关性。方法 通过反复动物接种和体外培养，观察动物肺转移状况，筛选高频转移细胞株，比较原发肿瘤和转移肿瘤的特征，并应用Northera-blot方法测定各类肿瘤细胞nm23 mRNA表达水平。结果 8株卵巢恶性肿瘤细胞中4株有较高转移潜能。多次培养接种可筛选出高频转移细胞亚群。测定各类细胞nm23 mRNA表达水平与肿瘤转移特性呈负相关。结论 由基因分子水平决定的肿瘤转移趋势在不同肿瘤种类及细胞亚群中有明显差异；卵巢癌中nm23 mRNA和蛋白的表达与其转移能力的降低有密切关系，可作为判定卵巢癌预后的敏感指标。     

卵巢癌高频转移细胞模型中nm23-H1基因表达的相关性研究

目的筛选高频转移卵巢恶性肿瘤细胞,研究不同转移潜能的细胞和 nm23的相关性。方法通过反复动物接种和体外培养,观察动物肺转移状况,筛选高频转移细胞株,比较原发肿瘤和转移肿瘤的特征,并应用 North-ern-blot 方法测定各类肿瘤细胞 nm23 mRNA 表达水平。结果 8株卵巢恶性肿瘤细胞中4株有较高转移潜能。多次培养接种可筛选出高频转移细胞亚群。测定各类细胞 nm23 mRNA 表达水平与肿瘤转移特性呈负相关。结论由基因分子水平决定的肿瘤转移趋势在不同肿瘤种类及细胞亚群中有明显差异;卵巢癌中 nm23 mRNA 和蛋白的表达与其转移能力的降低有密切关系,可作为判定卵巢癌预后的敏感指标。     

nm23-H1基因表达与卵巢癌转移的相关性

背景与目的：转移是卵巢癌治疗失败及患者死亡的首要原因。然而,目前对卵巢癌转移潜能的分子机制知之不多。本研究旨在筛选高频转移卵巢恶性肿瘤细胞,分析卵巢癌高频转移细胞模型中nm23．H1基因表达与肿瘤转移特性的相关性,为系统实验研究和临床实践提供依据。方法：通过反复动物接种和体外培养,观察动物肺转移状况,筛选高频转移细胞株,比较原发肿瘤和转移肿瘤的特征,并应用Northem blot和Westem blot方法测定各类肿瘤细胞nm23 mRNA和蛋白表达水平。结果：8株卵巢恶性肿瘤细胞中,4株有较高转移潜能,多次培养接种可筛选出高频转移细胞亚群。各类细胞nm23mRNA和蛋白表达水平与肿瘤转移特性呈负相关(r=0．96,P=0．0001)。结论：由基因分子水平决定的肿瘤转移趋势在不同肿瘤种类及细胞亚群中有明显差异;卵巢癌中nm23 mRNA和蛋白的表达与其转移能力的降低有密切关系,可作为判定卵巢癌预后的敏感指标。     

Metastatic characteristics of SY86B human gastric carcinoma in athymic nude mice

The invasive and metastatic behavior of SY86B human gastric carcinoma in nude mice were further studied. The results showed that this human cancer strain had a relatively high spontaneous metastatic rate (68.4%). The metastasis was related to the invasiveness of the transplanted tumor and tumor-bearing time. Since the metastasis developed from 2nd to 9th passages, it was considered that this model had a stable metastatic tendency. It was observed that the genetic background, age and sex difference of the recipient nude mice may not be associated with the metastasis. Aside from the high transplantable rate (100%) and spontaneous metastatic rate (68.4%), SY86B human gastric carcinoma strain still maintains the morphologic, immunologic, genetic and ultrastructural characteristics of the original tumor. All these make it a useful means in the study of diagnosis and treatment of cancer metastasis.     

A case of emergency operation for pedicle torsion of ovarian tumor during bevacizumab therapy

We report a case of torsion of ovarian carcinoma pedicle during chemotherapy with bevacizumab and FOLFIRI. A 47-year-old-female who had undergone laparoscopic-assisted sigmoidectomy for sigmoid cancer with multiple metastatic liver tumors, was treated with 3 courses of FOLFOX4 and 9 courses of FOLFIRI. Bevacizumab (BV) was started to administer with FOLFIRI since a CT scan had revealed that metastatic liver tumors had enlarged . Four days after final administration of BV (13th course), she presented lower abdominal pain. CT scan showed an enlarged right ovary sized 20 cm, but no findings suggesting bowel perforation; therefore we assumed her pain was due to enlarged ovarian tumor, which might be metastasis from sigmoid cancer. Increased oral morphine could not relieve her pain. Then, along with sufficient information about the risks of the operation caused by BV, we proposed an operation to remove her enlarged ovary. Day 16 after final administration of BV, we undertook a laparotomy by lower median incision, and found a twisted right adnex with large necrotic ovarian tumor. Pathologists diagnosed the ovarian tumor as primary ovarian cancer. No complication related to BV occurred, and she was moved to the ward for internal medicine 13 days after operation.     

Establishment of an ovarian metastasis model and possible involvement of E‐cadherin down‐regulation in the metastasis

Clinical observations of cases of ovarian metastasis suggest that there may be a unique mechanism underlying ovarian‐specific metastasis. This study was undertaken to establish an in vivo model of metastasis to the ovary, and to investigate the mechanism of ovarian‐specific metastasis. We examined the capacity for ovarian metastasis in eight different human carcinoma cell lines by implantation in female NOD/SCID mice transvenously and intraperitoneally. By transvenous inoculation, only RERF‐LC‐AI, a poorly differentiated carcinoma cell line, frequently demonstrated ovarian metastasis. By intraperitoneal inoculation, four of the eight cell lines (HGC27, MKN‐45, KATO‐III, and RERF‐LC‐AI) metastasized to the ovary. We compared E‐cadherin expression among ovarian metastatic cell lines and others. All of these four ovarian metastatic cell lines and HSKTC, a Krukenberg tumor cell line, showed E‐cadherin down‐regulation and others did not. E‐cadherin was then forcibly expressed in RERF‐LC‐AI, and inhibited ovarian metastasis completely. The capacity for metastasizing to the other organs was not affected by E‐cadherin expression. We also performed histological investigation of clinical ovarian‐metastatic tumor cases. About half of all ovarian‐metastatic tumor cases showed loss or reduction of E‐cadherin expression. These data suggest that E‐cadherin down‐regulation may be involved in ovarian‐specific metastasis. (Cancer Sci 2008; 99: 1933–1939)     

A GFP-labeled Human Colon Cancer Metastasis Model Featuring Surgical Orthotopic Implantation

Colorectal cancer has become a major disease threatening human health. To establish animal models thatexhibit the characteristics of human colorectal cancer will not only help to study the mechanisms underlying thegenesis and development effectively, but also provide ideal carriers for the screening of medicines and examiningtheir therapeutic effects. In this study, we established a stable, colon cancer nude mouse model highly expressinggreen fluorescent protein (GFP) for spontaneous metastasis after surgical orthotopic implantation (SOI). GFPlabeledcolon cancer models for metastasis after SOI were successfully established in all of 15 nude mice and therewere no surgery-related complications or deaths. In week 3, primary tumors expressing GFP were observed inall model animals under fluoroscopy and two metastatic tumors were monitored by fluorescent imaging at thesame time. The tumor volumes progressively increased with time. Seven out of 15 tumor transplanted mice diedand the major causes of death were intestinal obstruction and cachexia resulting from malignant tumor growth.Eight model animals survived at the end of the experiment, 6 of which had metastases (6 cases to mesentericlymph nodes, 4 hepatic, 2 pancreatic and 1 mediastinal lymph node). Our results indicate that our GFP-labeledcolon cancer orthotopic transplantation model is useful with a high success rate; the transplanted tumors exhibitsimilar biological properties to human colorectal cancer, and can be used for real-time, in vivo, non-invasive anddynamic observation and analysis of the growth and metastasis of tumor cells.     

Establishment and Characterization of a New Spontaneous Metastasis Model of Human Gastric Carcinoma in Nude Mice

A poorly differentiated medullary carcinoma of human stomach, designated HY–1, was successfully transplanted to nude mice by either the subcutaneous or intramuscular route for five generations. The transplanted tumor showed spontaneous lung metastases in nearly 100% of KSN and Balb/c female nude mice. There were over 20 visible lung metastatic nodules in KSN and Balb/c nude mice bearing tumors for over SO days. Immunostaining of type IV collagen and electron microscopy revealed that tumor cells were often in direct contact with basement membrane (BM) of tumor blood vessels in the primary tumor tissue. At the site of contact between tumor cells and vascular BM, focal disappearance of the BM, disruption of endothelial cells and entry of tumor cell clusters into vascular lumen were observed. Immunostaining of 72 kDa gelatinase/type IV collagenase demonstrated that tumor cells expressed this enzyme in their cytoplasm. These results suggest that spontaneous metastasis of this tumor may be partly due to a marked tendency to vascular invasion involving the following sequential events: tumor cell contact with vascular BM, BM degradation possibly by 72 kDa gelatinases and endothelial disruption. This model could be a useful tool for understanding the mechanism of hematogenous metastasis of human gastric cancer.     

Establishment and characterization of a new spontaneous metastasis model of human gastric carcinoma in nude mice.

A poorly differentiated medullary carcinoma of human stomach, designated HY-1, was successfully transplanted to nude mice by either the subcutaneous or intramuscular route for five generations. The transplanted tumor showed spontaneous lung metastases in nearly 100% of KSN and Balb/c female nude mice. There were over 20 visible lung metastatic nodules in KSN and Balb/c nude mice bearing tumors for over 80 days. Immunostaining of type IV collagen and electron microscopy revealed that tumor cells were often in direct contact with basement membrane (BM) of tumor blood vessels in the primary tumor tissue. At the site of contact between tumor cells and vascular BM, focal disappearance of the BM, disruption of endothelial cells and entry of tumor cell clusters into vascular lumen were observed. Immunostaining of 72 kDa gelatinase/type IV collagenase demonstrated that tumor cells expressed this enzyme in their cytoplasm. These results suggest that spontaneous metastasis of this tumor may be partly due to a marked tendency to vascular invasion involving the following sequential events: tumor cell contact with vascular BM, BM degradation possibly by 72 kDa gelatinases and endothelial disruption. This model could be a useful tool for understanding the mechanism of hematogenous metastasis of human gastric cancer.     

沉默乙酰肝素酶基因对胃癌生物学行为的影响

目的 探讨乙酰肝素酶(HPA)沉默对人胃癌裸鼠移植瘤生长、转移和血管形成的影响.方法 利用人胃癌SGC-7901细胞和HPA被沉默的SGC-7901-HPA-细胞,分别建立6只裸鼠皮下移植瘤模型,观察成瘤的时间、肿瘤生长速度和体积.应用逆转录聚合酶链反应(RT-PCR)和Western blot方法分别检测皮下移植瘤组织中HPA mRNA和蛋白的表达,应用免疫组织化学SP法检测皮下移植瘤组织的微血管密度(MVD).将皮下移植瘤细胞分别注射入6只裸鼠腹腔,建立腹腔转移瘤,并观察成瘤情况.结果 SGC-7901细胞和SGC-7901-HPA-细胞在裸鼠皮下均能生长出移植瘤,分别在接种后第4天后和第7天后出现肉眼可见的肿瘤,接种SGC-7901-HPA-细胞的裸鼠移植瘤生长较慢,MVD为(11.35±1.94)个/高倍视野,明显低于接种SGC-7901细胞组[(20.69±1.20)个/高倍视野,P＜0.05].接种SGC-7901-HPA-细胞与接种SGC-7901细胞的裸鼠皮下移植瘤组织相比,HPA mRNA和蛋白的表达均降低.由SGC-7901细胞皮下移植瘤组织建立腹腔转移瘤的裸鼠,有3只成瘤,在肝脏、大网膜、肠系膜、右肾形成了4处转移灶,且体积较大.而接种SGC-7901-HPA-细胞皮下移植瘤组织的裸鼠,仅有1只在肝脏和右肾形成了转移灶,而且瘤体较小.结论 HPA沉默后抑制了人胃癌在裸鼠体内的生长、转移和血管形成,HPA有可能成为预防和治疗胃癌的一个新靶H点.