Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.

Establishment of a mdr1 Multidrug Resistant Model of Orthotopic Transplantation of Liver Carcinoma on Nude Mice

To develop a new method of inducing mdrl multidrug resistance by establishinga nude mice model of orthotopic transplantation of liver carcinoma by sporadic abdominalchemotherapy at intervals. Methods: Hepatocellular carcinoma HepG2 cell was cultured and injectedsubcutaneously to form the tumor-supplying mice. The tumor bits from the tumor-supplying mice wereimplanted under the envelope of the mice liver and induced by abdominal chemotherapy withPharmorubicin. Physical examination, ultrasonography, spiral CT and operative inspection were usedto examine tumor progression. RT-PCR and immunohistochemistry were adopted to detect the expressionof mdr1-mRNA and its encoded protein P-gp protein (P-gp). Results: There was no operative dead, therate of implanting tumor successfully was 88% (22/25), the rate of implanting secondly successfullywas 100% (3/3), and the rate of inducing successfully was 80% (16/20). The expression of mdrl-mRNAand the P-gp in the inducing group was 23 folds and 13 folds in the control group respectively.Conclusion: We have established an in vivo model of mdr using nude mice transplanted with orthotopicliver neoplasm coupled to chemotherapy.

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.

Effects of endostatin on expression of vascular endothelial growth factor and its receptors and neovascularization in colonic carcinoma implanted in nude mice

AIM:To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS:Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma cell line to generate carcinoma and were randomly separated into two groups.Mice received injection of vehicle or endostatin every day for two weeks. After the tumor was harvested,the tumor volumes were determined,and the expressions of CD34,VEGF and FIk-1 were examined by immunohistochemical method. RESULTS:Tumor volume was significantly inhibited in the endostatin group(84.17%)and tumor weight was significantly inhibited in the endostatin group(0.197±0.049) compared to the control group(1.198±0.105)(F=22.56, P=0.001),microvessel density(MVD)was significantly decreased in the treated group(31.857±3.515)compared to the control group(100.143±4.290)(F=151.62,P<0.001). Furthermore,the expression of FIk-1 was significantly inhibited in the treated group(34.29%) ompared to the control group(8.57%)(X^2=13.745,P=0.001).However no significant decrease was observed in the expression of vascular endothelial growth factor(VEGF)between these two groups(X^2=0.119,P=0.730). CONCLUSION:Endostatin can inhibit tumor growth and angiogenesis by blocking Vegf/FIk-1 pathway.This experiment provides the theory basis for developing a new anti-carcinoma drug through studying the properties of anti-angiogenesis inhibitors.

Tumor radioimmunoimaging of chimeric antibody in nude mice with hepatoma xenograft

IM To study the radioimmunoimaging (RAII) using the human/mouse chimeric Ab to evaluate its targeting activity in animal models.METHODS To chimeric Ab was labeled with 131I. RAII was performed at different intervals after injection of radiolabeled Abs in nude mice with human hepatoma xenograft, and tissue distribution of radioactivity was measured. Comparison was made in the chimeric Ab between the single segment Ab and previous murine mAb against HBxAg.RESULTS The experimental objects developed tumorpositive image after 2 days of radiolabeled Abs injection, and the peak accumulation of radioactivity fell on the 7th day. The tumor/liver ratioactivity of the chimeric Ab, single segment Ab, antiHBx mAb, and the control group was 281±021, 244±016, 460±019, and 096±014, respectively.CONCLUSION The genetic engineering Abs have a considerable targeting activity which can be used as a novel humanized vector in the targeting treatment of liver cancer..

A nude mouse model of endometriosis and its biological behaviors

Endometriosis （ EM ） as a common and intractable gynecological disease is characterized by unknown etiology and complex pathologic changes. Many factors of the disease are uncertain at the molecular level and it is difficult to study clinically. In this study, we attempted to establish a nude mice model of EM for dynamical observation of the genesis and development of the disease, morphological changes in tissue, and biological behaviors.

Translational pancreatic cancer research:a comparative study on patient-derived xenograft models

AIM To assess the viability of orthotopic and heterotopic patient-derived pancreatic cancer xenografts implanted into nude mice.METHODS This study presents a prospective experimental analytical follow-up of the development of tumours in mice upon implantation of human pancreatic adenocarcinoma samples. Specimens were obtained surgically from patients with a pathological diagnosis of pancreatic adenocarcinoma. Tumour samples from pancreatic cancer patients were transplanted into nude mice in three different locations(intraperitoneal, subcutaneous and pancreatic). Histological analysis(haematoxylin-eosin and Masson's trichrome staining) and immunohistochemical assessment of apoptosis(TUNEL), proliferation(Ki-67), angiogenesis(CD31) and fibrogenesis(α-SMA) were performed. When a tumour xenograft reached the target size, it was reimplanted in a new nude mouse. Three sequential tumour xenograft generations were generated(F1, F2 and F3).RESULTS The overall tumour engraftment rate was 61.1%. The subcutaneous model was most effective in terms of tissue growth(69.9%), followed by intraperitoneal(57.6%) and pancreatic(55%) models. Tumour development was faster in the subcutaneous model(17.7 ± 2.6 wk) compared with the pancreatic(23.1 ± 2.3 wk) and intraperitoneal(25.0 ± 2.7 wk) models(P = 0.064). There was a progressive increase in the tumour engraftment rate over successive generations for all three models(F1 28.1% vs F2 71.4% vs F3 80.9%, P < 0.001). There were no significant differences in tumour xenograft differentiation and cell proliferation between human samples and the three experimental models among the sequential generations of tumour xenografts. However, a progressive decrease in fibrosis, fibrogenesis, tumour vascularisation and apoptosis was observed in the three experimental models compared with the human samples. All three pancreatic patient-derived xenograft models presented similar histological and immunohistochemical characteristics.CONCLUSION In our experience, the faster development andgreatest number of viable xenografts could make the subcutaneous model the best option for experimentation in pancreatic cancer.

人源异种皮下注射移植法建立子宫内膜异位症纤维化裸鼠模型的评价

目的评估人源子宫内膜异位症纤维化裸鼠模型建立的可行性,确定子宫内膜间充质干细胞是否参与诱导子宫内膜异位症纤维化。方法收集人源子宫内膜标本4例,采用皮下注射的方式1∶3移植到12只BABL/c裸鼠体内。记录病灶皮表形态及体积变化,移植后第15天处死裸鼠,观察病灶形态及其与腹壁周围粘连情况,HE染色评判造模结果,Masson染色评定纤维化程度,免疫荧光追踪子宫内膜间充质干细胞在子宫内膜异位症纤维化进程中的作用。结果裸鼠异位病灶体积随时间增长,呈局限性、囊泡样改变,与腹壁粘连紧密,镜下可见子宫内膜样腺体、胶原纤维沉积,造模成功率为83.4%,造模后病灶胶原纤维容积分数显著升高(P<0.01),共聚焦成像提示子宫内膜间充质干细胞(SUSD2^(+))可在体内向肌成纤维细胞(α-SMA^(+))分化。结论人源异种皮下注射移植的方法建立的裸鼠模型符合子宫内膜异位症纤维化的病变特点,操作简单,可行性高,此外体内观察到子宫内膜间充质干细胞参与诱导子宫内膜异位症纤维化形成,为进一步探究其发病机制提供较好的模型参考。

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.

AEG-1对裸鼠肝癌模型中肝癌细胞生长及肺转移的影响

目的研究过表达和沉默AEG-1基因对肝癌细胞生长的影响,以及AEG-1基因在调节肝癌细胞定向肺转移中的作用。方法分别以携带过表达AEG-1序列及对照基因序列的慢病毒(lentivirus)转染SMMC-7721肝癌细胞株(SMMC-7721-AEG-1-L;SMMC-7721-control-L);以携带shRNA AEG-1及对照shRNA质粒(plasmid)转染SMMC-7721细胞株(SMMC-7721-shAEG-1-P;SMMC-7721-control-P);使用实时定量PCR和Westen blot检测AEG-1的表达;随后使用荧光素酶基因慢病毒包装颗粒转染上述4种稳定转染细胞株。使用上述细胞株分别建立3种裸鼠肝癌模型:皮下移植瘤模型,原位移植瘤模型和血行播散模型;每种细胞株每一模型5只Balb-c裸鼠,共60只。建立皮下移植瘤模型,观测肿瘤的生长情况并绘制肝细胞肿瘤生长曲线;建立肝癌原位移植瘤和血行播散模型,采用生物发光活体成像技术及组织病理学方法监测造模成功率及肿瘤在肝内、肝外转移情况。结果通过肝癌皮下移植瘤模型可观察到AEG-1过表达组肿瘤体积显著高于对照组,且裸鼠肝脏组织出现弥漫性侵袭转移灶;在肝癌原位移植瘤和血行播散模型中可观察到AEG-1过表达/沉默可以导致肝癌细胞肝内转移、肺转移率和转移灶数量显著增高/降低。结论过表达AEG-1基因可促进肝癌细胞生长以及定向肺转移,沉默AEG-1基因抑制肝癌细胞生长及定向肺转移。

肺癌颅脑转移动物模型建立的比较

目的通过左心室注射、尾静脉注射、胸腔注射3种方法,建立肺癌颅脑转移的动物模型。方法18只BALB/c NUDE裸鼠随机分为三组,分别经左心室注射、尾静脉注射、胸腔注射途径,接种具有颅脑转移潜能的人肺腺癌PC-9细胞(1×10^(6)/0.1 mL),接种后观察裸鼠状态,在裸鼠出现恶病质时处死;解剖裸鼠,肉眼观察肺和颅脑的转移情况,并对其进行HE染色处理。结果左心室注射组:裸鼠注射后18 d开始出现体质量下降,逐步出现恶病质,并于27 d将裸鼠全部处死;解剖观察肺、颅脑无异常;HE染色证实,肿瘤病灶并未在它们的肺部发现;6只裸鼠均有共济失调、偏瘫、跛行的症状,是颅脑转移的病灶引起的。尾静脉注射组:注射后55 d裸鼠体质量开始减轻,92 d开始出现恶病质,处死于112 d;解剖观察肺、颅脑无异常;HE染色证实4只裸鼠身上出现肺部肿瘤灶;6只裸鼠均未发现颅脑转移。胸腔注射组:裸鼠于注射后22 d出现胸壁瘤结;35 d开始有消瘦,恶病质渐起,42 d处死;解剖观察肺部、胸腔见多发肿瘤结节,并胸膜、肋骨、脊柱浸润;HE染色证实6只裸鼠全部表现为肺部转移;6只裸鼠均未发现颅脑转移。结论左心室注射组脑转移率为100%,肺癌颅脑转移动物模型建立的可靠方法是经左心室注射途径。

人乳腺癌裸鼠移植模型的建立

目的 :建立人乳腺癌裸鼠移植模型 ,并探讨其部分生物学特性。方法 :采用雌激素受体阳性的 MCF- 7人乳腺癌细胞株 ,接种于 2 0只裸小鼠右侧胸壁乳垫下 ,移植细胞总数为 1× 10 6 /只。观察肿块生长情况 ,至 90天处死荷瘤鼠 ,切除肿块作病理切片 ,角蛋白19( KT19) m RNA、雌激素受体 ( ER)检测。结果 :接种后第 10天在接种部位可见结节 ,肿瘤移植成功率 (成瘤率 )为 95 %( 19/2 0 )。切除肿块平均直径为 ( 14± 3 ) mm,平均重量为 1.4 g,病理学检查为浸润性导管癌 ,RT- PCR检测表达人角蛋白 19( KT19)、ER阳性。结论 :该方法建立的人乳腺癌裸鼠移植模型 ,成功率高 ,肿瘤可部分保持人乳腺癌生物学特性 。

子宫内膜异位症裸鼠模型子宫内膜的雌、孕激素受体表达

目的:建立子宫内膜异位症裸鼠模型,观察子宫内膜的雌激素受体(ERs)、孕激素受体(PRs)及其亚型的表达情况,为进一步探讨内异症子宫内膜孕激素抵抗的发生机制提供研究模型。方法:建立子宫内膜异位症裸鼠模型,分别于造模后1月、2月、3月取裸鼠子宫内膜,利用RT-qPCR和Western blot法检测其孕激素受体(PRA、PRB)和雌激素受体(ERα、ERβ)表达情况。结果:与对照组相比,内异症组裸鼠内膜的PRB和ERα表达降低,在造模后3月最明显;PR和ERβ表达升高,PRB/PRA和ERα/ERβ比值降低,均在造模后3月最明显。结论:子宫内膜异位症裸鼠模型是研究内异症的理想模型之一,造模后的裸鼠内膜存在孕激素抵抗。

三种人卵巢癌动物模型的生物学特性比较

目的:建立人卵巢癌裸小鼠皮下移植瘤,腹水瘤和原位移植瘤模型,观察比较其生物学特性,为卵巢癌的理论研究和选择临床模型选择提供参考。方法:利用卵巢癌细胞株SW626先制备裸小鼠皮下移植瘤模型,再将瘤组织条植入裸小鼠一侧卵巢内,建立人卵巢癌的原位移植模型,将该细胞悬液注入小鼠腹腔建立腹水瘤模型。用组织病理学,电镜和流式细胞仪DNA含量及染色体核型分析鉴定模型,并比较3种模型在生物学特性上的差异。结果:3种模型的瘤细胞与细胞株在形态和结构上一致,腹水瘤模型自然生存率明显短于其它两种模型,后两者无明显差异。在生物学特性上各具特点,其中原位移植瘤模型中肿瘤的生长和转移完全模拟人卵巢癌的临床过程。结论:3种模型从不同层面展现了人卵巢癌的生物学特点,其中原位移植瘤模型是研究卵巢癌的转移基础和评定抗癌药物药效更客现的模型。

ErbB2高表达的人乳腺癌原位移植瘤裸鼠模型的建立

目的采用人乳腺癌细胞株BT-474建立稳定高表达ErbB2的人乳腺癌裸鼠移植瘤模型,选择合适的模型制作方法。方法实验前1d在36只裸鼠颈部皮下埋植0.5mg17β雌二醇缓释片,并随机分为3组:原位组将5×106个BT-474细胞接种于裸鼠左侧第二乳房垫内,胁部皮下组将同样数量的BT-474细胞接种于裸鼠左侧胁部皮下,腋窝组则接种于裸鼠左侧腋窝内;观察3组裸鼠荷瘤情况及移植瘤病理组织学特点,并采用免疫组化方法动态监测移植瘤的ErbB2表达情况。结果 3组裸鼠的荷瘤率均为100%,都保持了原发肿瘤高表达ErbB2的特点;和两组对照相比,原位组肿瘤形状、生长速度及细胞增殖与血管生成等生物学特征一致性更强;此外,原位组的肿瘤对治疗药物处理也有良好反应。结论 BT-474细胞接种于乳房垫内比胁部皮下及腋窝建立的裸鼠移植瘤模型更适于进行抗ErbB2抗体等药物的药理学研究。

VEGF在人子宫内膜异位症裸鼠模型组织中的表达及意义

目的建立人子宫内膜异位症(EMs)裸鼠模型,检测血管内皮生长因子(VEGF)在正常在位内膜及裸鼠异位内膜的表达,探讨其在EMs发生发展中的作用。方法分别采用皮下种植和腹腔注射的方法,建立EMs裸鼠模型,光镜下观察异位病灶的形态学特点,并采用免疫组化法检测正常在位内膜及裸鼠异位内膜中VEGF蛋白表达水平。结果实验裸鼠共20只,皮下种植组10只,8只成模,腹腔注射组10只,7只成模,异位病灶在光镜下呈现增生期特点;正常在位内膜10例,VEGF蛋白在异位内膜中的表达有增多趋势(t=2.632,P<0.05)。结论成功建立人EMs裸鼠皮下种植及腹腔注射模型,检测到VEGF表达较正常在位内膜增多,该模型可用于进行人EMs血管生成及抗血管生成治疗的研究模型。

人子宫内膜异位症裸鼠模型的建立及雌、孕激素受体的表达

目的 为子宫内膜异位症 (EM)的临床研究提供实验动物模型。方法 实验组 :取EM和正常育龄妇女分泌晚期子宫内膜 ,剪碎 ,随机置入裸鼠盆腹腔 ,实验Ⅰ组切除卵巢 ,实验Ⅱ组不切除卵巢 ;对照组 :同法置入人大网膜。术后给雌激素支持。各组裸鼠分别于 5、10、15、2 0、2 5、3 0d脱颈椎处死 ,观察异位病灶生长情况。免疫组化SP法检测异位病灶雌、孕激素受体表达。结果 子宫内膜种植 5d即见异位病灶牢固粘附 ,主要发生在腹壁(56% )、膀胱旁脂肪 (2 1% )等部位 ,可见腺体细胞及散在间质细胞 ,时间越长 ,病灶与裸鼠组织融合越明显 ,盆腹腔粘连越重。用EM患者和正常育龄妇女内膜移植的异位病灶无差别 ,实验Ⅰ组和实验Ⅱ组的裸鼠所形成的异位病灶亦无差别 ,它们的雌、孕激素受体表达未显示差别 ,多数呈弱表达。结论 裸鼠做为EM早期动物模型科学、简便、可行。

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。

快速建立裸鼠胶质瘤模型的实验研究

目的:探讨快速建立裸鼠C6型脑胶质瘤动物模型的方法及在科研实验中应用的可行性。方法:将C6型鼠胶质瘤细胞悬液分别接种至雌雄裸鼠皮下,或用瘤组织块行异体移植至雌雄裸鼠皮下,观察裸鼠皮下移植瘤长出时间,并研究肿瘤的病理组织学及免疫组织化学特征。结果:(1)两种方法所建肿瘤模型其成功率均为100%,肿瘤生长状况良好,肿瘤病理组织学及免疫组织化学检查均符合胶质瘤细胞的形态学特征;(2)采用细胞接种法建立裸鼠脑胶质瘤模型所需的时间长,在雌雄裸鼠皮下生长速度不一,差别有统计学意义,P<0.05;(3)采用瘤组织块行异体移植法建立的肿瘤模型,其所需时间均较细胞接种法明显缩短,雌雄裸鼠肿瘤组织块生长速度基本相同,差别没有统计学意义,P>0.05。结论:(1)利用肿瘤组织块移植的方法可快速建立裸鼠皮下移植瘤模型,并能作为研究胶质瘤发病机制、生物学特性以及基因治疗的可靠动物模型;(2)利用肿瘤组织块异体移植的方法,可以缩短肿瘤研究中用于建立动物模型的时间,节约经济成本。

肺癌转移瘤动物模型的建立

目的通过尾静脉注射,建立一种符合临床特征的肺腺癌转移瘤动物模型,为下一步的肺腺癌转移机制的研究提供可靠的实验造模方法。方法取对数生长期的A549细胞,11只SPF级、4~6周龄BALB/c裸鼠,分别以1×106个细胞/只注射入裸鼠尾静脉。接种后每天观察小鼠状态。分别于接种肿瘤细胞后第4、5、6、7周随机处死2只,余3只小鼠处于濒死状态时处死。解剖小鼠,观察肺部有无转移、转移结节的数目及全身其他器官的转移情况,并做病理取材,HE染色观察。结果注射过程中小鼠均存活。未处死的3只分别于第11、13、14周出现恶液质。第4周肺部未见转移结节;第5周出现镜下肺部转移结节;第6周肉眼可见肺部转移结节;第7周转移结节数增多;第11周出现纵隔淋巴结转移。第11、13、14周出现肺部结构大量破坏,弥漫性的肿瘤细胞浸润,出现淋巴结浸润,病理证实为腺癌。结论通过尾静脉注射A549细胞可以成功建立人肺腺癌转移瘤模型。