In vivo metastasis of human tumor transplant in T and NK cell immune-deficient mice

Beige-nude mice with combined T and NK cell deficiency were produced by introducing "nu" gene into the beige mice of C57BL/6 background. The beige-nude mice were characterized by morphological features, level and activity of the NK cell in spleen and peripheral blood. NK cell level of the beige-nude mice was higher than the beige mice and similar to the ordinary nude mice though its NK cell activity was markedly lower than the nude mice. Human lung adenocarcinoma (PAa) was transplanted subcutaneously to the beige-nude mice and nude mice of BALB/cA background. No metastasis was found in BALB/cA nude mice (0/7), while in beige-nude mice, spontaneous lymph node metastasis rate was 36.3% (4/11). It should be noted that the transplanted PAa metastasized to lungs, which had never been seen in BALB/cA nude mice in previous experiments. The increase of metastatic rate is correlated with the relative low NK cell activity of the beige-nude mice. Our observation is that the beige-nude mice could be a suitable experimental model for in vivo study on metastatic behavior of the human tumors.     

Development of androgen-independent spindle cell tumors from androgen-dependent medullary Shionogi carcinoma 115 in androgen-depleted nude mice.

When Shionogi carcinoma 115 (SC115, undifferentiated medullary carcinoma showing compact cell pattern and containing androgen receptor) was transplanted into male and female DS mice, it grew only in males. In contrast to this strict androgen dependency in DS hosts, tumors composed of spindle-shaped cells appeared in more than 80% of cases when SC115 tumor was inoculated into female or castrated male nude athymic (BALB/c-nu/nu) recipients. These spindle cell tumors neither contained cytosol androgen receptor nor showed biologically defined androgen dependency. As spindle cell tumors could be serially transplanted in DS mice but not in BALB/c-+/+ mice and as the original SC115 (medullary carcinoma showing a compact cell pattern) tumor and the spindle cell tumor had many identical chromosome abnormalities, these two types of tumors seem to have a common origin in spite of their morphological, biochemical, and biological differences. Since spindle cells could not be detected histologically in SC115 tumors maintained in intact male DS mice, the present results seem to suggest that SC115 cells may change their morphological, biochemical, and biological characteristics within one passage in androgen-depleted nude athymic mice.     

基质降解酶u-PA,PAI-1与人肺癌细胞浸润转移的相关关系的研究

观察体外细胞株ＰＧ、ＰＡａ细胞的ｕ－ＰＡ，ＰＡＩ－１活性和抗原表达，结果发现ＰＧ、ＰＡａ细胞均能够产生ＰＡ及ＰＡＩ，免疫细胞化学法定位ＰＡ主要为ｕ－ＰＡ，ＰＡＩ主要为ＰＡＩ－１。在ＰＧ为ｕ－ＰＡ高表达，而ＰＡａ为低ｕ－ＰＡ表达，ＰＡＩ－１在ＰＧ、ＰＡａ细胞中均高表达。对ＰＧ瘤细胞在ＢＡＬＢ／ＣＡ裸鼠皮下种植性瘤组织的免疫组化结果显示，ｕ－ＰＡ阳性～强阳性表达，ＰＡＩ－１为阴性表达。说明ｕ－ＰＡ是ＰＧ、ＰＡａ高低不同的转移能力的重要机制之一。     

Early tumor growth in metastatic organs influenced by the microenvironment is an important factor which provides organ specificity of colon cancer metastasis

We have previously demonstrated that liver metastases in nude mice and lung metastases in nude rats occurred specifically, when KM12SM human colon carcinoma cells were inoculated orthotopically into the cecal wall of nude mice and rats. To clarify the relationship between the tumor growth potential in the metastatic organs and the metastatic organ preference in these two metastatic models, we have evaluated the in vitro cell growth activities affected by the organ conditioned medium (CM) from the liver and lung, and the in vivo growth activities of the ectopic implanted tumors in the liver and lung. The tumorigenicity of the ectopic implanted tumors was 100% in mouse liver, 33% in rat liver, 50% in mouse lung, and 75% in rat lung. The crude liver CM of the animals showed inhibitory activities for KM12SM cell growth in a dosage-dependent manner, and the crude lung CM stimulated KM12SM cell growth. The liver CM of nude mice inhibited the KM12SM cell growth more strongly compared with the CM of nude rats, and the lung CM of nude rats was more strongly stimulated compared with the CM of nude mice. The liver CM of nude mice had non-heparin binding factors, which stimulated or inhibited KM12SM cell growth, in a molecular weight range of 50 to 100 kDa. By contrast, the liver CM of nude rats showed no growth stimulating activity for KM12SM cells. These results suggest that the metastatic organ specificity of KM12SM cells may depend on the early tumor growth influenced by the microenvironment in metastatic organs.     

Cell kinetics and chemosensitivity of human carcinomas serially transplanted into nude mice

Six human carcinoma xenografts serially transplanted into nude mice were used for the study of chemosensitivity and cell kinetics. Three gastric carcinomas (St-4, St-40 and H-111), two colon carcinomas (Co-3 and Co-4) and one breast carcinoma (MX-1) were inoculated into the subcutaneous tissue of BALB/cA nude mice. The maximum tolerable doses of mitomycin C (MMC), adriamycin (ADM), cyclophosphamide (CPA) and 5-fluorouracil (5-FU) were administered when the tumor weights reached 100-300 mg. The response rates of the tumor to these drugs were found to be 3/6 for MMC, 2/6 for 5-FU and 1/6 for ADM and CPA. Percent labeled mitosis curves obtained from 3H-thymidine pulse labeling were analyzed by the method of Quastler and Sherman. It was found that the antitumor effect of MMC was closely correlated with the growth fractions of the tumors (r = -0.98, P less than 0.001), and it appeared that the tumor cells were more sensitive to MMC in the resting stages during the proliferating phase than in the other cell cycle phases. Cell kinetics is considered to be an important factor in determining chemosensitivity, and the system of human tumor xenografts-nude mice seems to be a suitable experimental model for investigating the correlation between cell kinetics and chemosensitivity in vivo.     

ABCE1-GFP肺腺癌动物模型的建立及免疫组化检测

目的:建立ABCE1-GFP（+）肺腺癌裸鼠动物模型,并进行免疫组化检测,为进一步探索ABCE1作为诊断治疗非小细胞肺癌靶点提供有用的工具。方法:应用前期实验中构建的稳定转染重组质粒pEGFP-C1-ABCE1及空载体pEGFP-C1的LTEP-a-2肺腺癌细胞株,接种于裸鼠背部侧翼至腋窝皮下,建立ABCE1-GFP肺腺癌动物模型。进行成瘤情况观察,并予以倒置荧光显微镜系统呈像。取材后进行免疫组织化学检测。结果:稳转细胞系皮下接种裸鼠后,全部成瘤,过表达组肿瘤体积大于空载体及对照组（P＜0.05）。过表达组肿瘤在荧光显微镜系统下可稳定表达绿色荧光,成功构建了ABCE1-GFP（+）肺腺癌皮下成瘤的裸鼠动物模型。免疫组织化学检测亦验证这一模型的确立。结论:稳定转染LTEP-a-2肺腺癌细胞可以用来成功建立皮下ABCE1-GFP（+）肺腺癌成瘤模型,是较为理想研究ABCE1在非小细胞肺癌中的基因功能的动物模型。     

Effects of progesterone on human endometrial carcinoma cells in vivo and in vitro

The effects of progesterone on the growth and differentiation of human endometrial adenocarcinoma cells (cell lines SNG-P and SNG-M derived from primary and metastatic tumors, respectively) were assessed in vitro and in vivo. Progesterone suppressed their growth and induced cell differentiation in vitro. The suppressive effect of progesterone was stronger in the primary tumor cells than in the metastatic ones. Progesterone produced morphologic changes such as multinucleation, multinucleolation, vacuolation, extensive Golgi apparatus, and papillary arrangement of cells. The cells were transplanted sc into nude BALB/c mice where they produced undifferentiated adenocarcinomas in untreated mice and well-differentiated adenocarcinomas in progesterone-treated ones. Progesterone reduced tumor growth and decreased transplantability in nude mice. This hormone produced no change in the distribution of the chromosome numbers or in the karyology.     

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.     

ANTITUMOR EFFECTS OF HUMAN IL-15 GENE MODIFIED LUNG CANCER CELL LINE

ＡＮＴＩＴＵＭＯＲＥＦＦＥＣＴＳＯＦＨＵＭＡＮＩＬ－１５ＧＥＮＥＭＯＤＩＦＩＥＤＬＵＮＧＣＡＮＣＥＲＣＥＬＬＬＩＮＥＳｈｅｎＹｏｎｇｑｕａｎ１沈永泉ＣｕｉＬｉａｎｘｉａｎ２崔莲仙ＨｅＷｅｉ１何维ＸｕｅＬｉ１薛莉ＢａＤｅｎｉａｎ１巴德年１Ｉｎｓｔ...     

Establishment of a human chorionic gonadotropin-producing human gastric carcinoma in nude mice

A human chorionic gonadotropin (HCG)-producing gastric carcinoma was transplanted into BALB/c nu/nu nude mice. The original tumor tissue had been obtained by gastrectomy from a 55-year-old patient with gastric carcinoma. The tumor was transplanted serially in nude mice, and its doubling time was stable to approximately 12 days. A positive correlation was observed between serum HCG level and tumor weight. The serum HCG level of the mice and the histology of the tumor weight. The serum HCG level of the mice and the histology of the tumor, papillary adenocarcinoma with partial poorly differentiated adenocarcinoma, did not change on serial transfers. HCG was positively stained by an immunochemical technique only in the site of the poorly differentiated adenocarcinoma, while staining was negative in the site of the papillary adenocarcinoma, which constituted most of the tumor. These results indicate that this tumor in nude mice will be useful not only as a therapeutic experimental system but also for studying the potential malignancy of HCG-producing cancer cells.     

二烯丙基二硫对人胃癌细胞裸鼠移植瘤的抗肿瘤作用

背景与目的:既往研究发现二烯丙基二硫(diallyldisulfide,DADS)在体外可抑制多种肿瘤细胞生长,但在体内抗肿瘤作用的研究报道较少。本实验旨在探讨DADS对人胃癌细胞移植瘤在BALB/C裸鼠体内生长的影响。方法:未经药物处理和经30mg/LDADS处理1天的胃癌细胞MGC803接种于裸鼠皮下;观察体外DADS处理MGC803细胞裸鼠移植瘤的成瘤情况和未处理MGC803细胞移植瘤成瘤后腹腔注射DADS对胃癌移植瘤在BALB/c裸鼠体内生长情况的影响。Westernblot检测瘤组织中增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达情况。结果:30mg/LDADS处理的MGC803细胞移植裸鼠体内无一成瘤。荷瘤裸鼠腹腔注射DADS剂量为50、100和200mg/kg时的抑瘤率分别为27.8%、66.1%和73.0%,同时可抑制移植瘤癌细胞PCNA的表达。结论:DADS可明显降低胃癌细胞裸鼠移植瘤的成瘤性,并对移植瘤生长有明显抑制作用。     

Levels of plasmatic fibronectin in mice bearing adenocarcinomas of different metastasizing ability

The levels of fibronectin (FN) were assayed in plasma of BALB/c mice subcutaneously inoculated with a mammary adenocarcinoma of moderate metastatic ability (M3) and a related variant tumor with higher metastasizing potential (MM3). The mean plasmatic FN concentration increased in parallel with increased M3 and MM3 size and weight. Highest and earliest FN increases were observed in mice inoculated with the rapidly growing M3 tumor. A strong correlation between the level of plasma fibronectin and the number of lung metastases was only found in MM3 inoculated mice. Plasma fibronectin level is a good biological marker of tumoral growth rate in these adenocarcinoma tumors, but its role in the metastatic process warrants investigation.     

A cholangiocellular carcinoma nude mouse strain showing histologic alteration from adenocarcinoma to squamous cell carcinoma.

BACKGROUND. Adenosquamous carcinoma and squamous cell carcinoma (SCC) occur rarely in the liver compared with adenocarcinoma, and the histogenesis and biologic behaviors of these tumors remain unknown. The authors addressed these issues in the current article. METHODS. A specimen aseptically obtained from the surgically resected cholangiocellular carcinoma (CCC) was cut into pieces and inoculated into the back of a nude mouse, bilaterally. The developed tumors were resected and serially transplanted into nude mice. The morphologic features and growth kinetics of the nude mouse tumors at different passages were compared. RESULTS. The authors established a new human CCC nude mouse strain, designated nuKMC-2, from a 64-year-old woman. The original tumor of the patient showed the features of moderately differentiated tubular adenocarcinoma with small sheet-like arrangement of polygonal cells. The initial tumor developed in a nude mouse showed morphologic features similar to the original tumor. With the serial transplantation to nude mice, the components of tubular adenocarcinoma diminished, and all of the nuKMC-2 was replaced by SCC. Doubling times of nuKMC-2 at the 5th and 11th passages were 9.9 and 7.4 days, respectively, which suggested that the tumor with squamous components were more aggressive biologically than the adenocarcinoma. CONCLUSIONS. The results suggested that adenosquamous carcinoma might be a transitional form from adenocarcinoma to SCC and that some of the primary hepatic SCC might originate from adenocarcinomas.     

ELAM-1在鼻咽癌裸鼠移植瘤中的表达及其与转移的关系

目的 建立裸鼠鼻咽癌转移模型并探讨 E-选择素（ELAM-1）与鼻咽癌转移的相关性。方法 将鼻咽癌5-8F细胞悬液注射于裸鼠左后肢爪垫，观察裸鼠状态、成瘤情况并测量裸鼠体重及移植瘤长短径；采用连续病理切片苏木精-伊红染色观察移植瘤及转移情况，将16只人鼻咽癌荷瘤裸鼠分为转移组和非转移组；采用免疫组织化学法检测两组移植瘤组织中ELAM-1的表达。 结果 16只裸鼠均成瘤，成瘤率为100.0%，其中10只裸鼠出现转移瘤，转移率为62.5%。建模前，两组裸鼠体重差异无统计学意义［（13.83±0.56）g vs （14.62±0.30） g，t=1.026，P=0.071］。建模后4~7周，裸鼠瘤体体积呈指数增长，且转移组移植瘤增长速度较非转移组快，非转移组裸鼠瘤体体积小于转移组［（198.91 ± 163.29） mm³ vs （268.76 ±174.31） mm³，t=4.376，P=0.005］。ELAM-1在鼻咽癌裸鼠移植瘤、淋巴结转移灶及远处转移灶中的表达均为阳性，主要表达于细胞膜。转移组移植瘤光密度值高于非转移组（0.4497±0.0705 vs 0.0435±0.0082，t=4.388，P＝0.001）。结论 本研究成功构建稳定性好、转移率高的鼻咽癌裸鼠移植瘤转移模型，且ELAM-1在裸鼠移植瘤中高表达，可促进鼻咽癌裸鼠移植瘤生长和转移。     

GOLM1调控PI3K/AKT/mTOR信号转导通路促进肺腺癌细胞增殖、侵袭和迁移的机制研究

背景与目的：高尔基体膜蛋白1(Golgi membrane protein 1,GOLM1)在肺腺癌中发挥促癌作用,但对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制尚不明确。探究GOLM1对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制。方法：选取2019年4月—2021年4月在克拉玛依市中心医院行手术切除的肺腺癌患者的癌组织及相应癌旁组织标本各90例,采用免疫组织化学法检测肺腺癌组织和癌旁组织中GOLM1的表达,并分析肺腺癌组织中GOLM1表达与临床病理学特征的关系。采用蛋白质印迹法（Western blot）检测人肺上皮细胞系BEAS-2B及人肺腺癌H460、A549、PG49、H1299细胞系中GOLM1的表达。取对数生长期的肺腺癌A549细胞,随机分为空白组（细胞未转染）、GOLM1小干扰RNA阴性对照（small interfering RNA negative control,si-NC）组（细胞转染si-NC）、GOLM1小干扰RNA(GOLM1 small interferingRNA,si-GOLM1)组（细胞转染si-GOLM1）、胰岛素样生长因子-1(insulin...     

Erythropoietin-induced polycythemia in athymic mice following transplantation of a human renal carcinoma cell line

An established cloned human renal carcinoma line RC-1, which has been continuously maintained in culture for several years and which produces erythropoietin, was injected s.c. into BALB/c athymic mice and produced tumors. Tumorigenicity was directly correlated with the number of RC-1 cells inoculated. Tumor cell histology resembled the original patient-derived tumor. Tumor-bearing mice developed hepatosplenomegaly and significant reticulocytosis with elevated hemoglobin and hematocrit values that were proportional to tumor mass. In addition, red cell mass and blood volume of nude mice increased over 100% as compared to control mice or to animals bearing nonrelevant neoplasms. Large amounts of immunoreactive erythropoietin could be extracted from the nude mouse RC-1 tumors. These results indicate that the RC-1 cell line is tumorigenic and produces biologically active erythropoietin in vivo in athymic mouse hosts, thus providing a reproducible model to study ectopic erythropoietin production and its regulation in vivo.     

Characterization of F3II,a sarcomatoid mammary carcinoma cell line originated from a clonal subpopulation of a mouse adenocarcinoma

We characterized a new mammary tumor cell line, F3II, previously established in vitro from a clonal subpopulation of the BALB/c transplantable mammary adenocarcinoma M3, moderately metastatic to lung. The F3II cell line has been passaged >50 times. It has grown as elongated cells adherent to the bottom of the flask. Cytogenetic studies showed that F3II cultures were nearly triploid. Tumor cells expressed fibronectin and showed high levels of cell-surface urokinase, a key protease in invasion and metastasis. F3II cells grew as poorly differentiated, spindle-cell carcinoma tumors (sarcomatoid carcinomas) with a prominent local invasiveness, a high angiogenic response, and a 90–100% incidence of lung metastases when inoculated s.c. into syngeneic mice. Ultrastructural and immunocytochemical analysis revealed characteristic features of carcinomas. Our data suggest that F3II is less differentiated and more aggressive than the original tumor line, supporting the notion that mammary carcinomas are heterogeneous neoplasms and contain subpopulations with diverse biologic behavior. The F3II mouse mammary sarcomatoid carcinoma line is a suitable model to examine antiinvasive, antiangiogenic, and antimetastatic agents. © 1996 Wiley-Liss, Inc.     

Transplantation of human tumors in nude mice.

Ninety-one human tumors, including various common carcinomas, low-grade malignant tumors, and benign tumors, were transplanted into athymic nude mice. Tumor take was confirmed histologically for 22 neoplasms at the initial transplantation, and 14 serially transplantable tumors were established, including some hitherto unestablished or unreported, such as lung and hepatic cell carcinomas. Among the 91 tumors were 21, 14, and 13 carcinomas of the lung, stomach, and breast, respectively. Transplantability was highest in lung carcinomas (10/21), followed by gastric carcinomas (2/14) and breast carcinomas (1/13). Morphology of original tumors was retained well in most transplanted tumors, but desmoplastic or scirrhous tumors, such as gastric and breast carcinomas, tended to become medullary with a decrease in amount of tumor stroma. The ability to produce mucin in gastric carcinomas or melanin in malignant melanoma was maintained in serially transplantable tumors. In addition, ectopic production of adrenocorticotropin and beta melanocyte-stimulating hormone continued in a transplanted small cell carcinoma of the lung. Preliminary results were obtained on hormone dependency of the transplantable breast carcinoma and on alpha1-fetoprotein in the transplantable hepatic cell carcinoma.     

Induction of other differentiation stages in neoplastic epithelial duct and myoepithelial cells from the human salivary gland grown in athymic nude mice

The interaction between two cell lines derived from the human salivary gland (HSG), neoplastic epithelial duct HSG cells and myoepithelial human pleomorphic adenoma (HPA) cells, was studied morphologically and immunohistochemically in nude mice tumors produced by inoculation of HSG and HPA cells. Transplantation of HSG cells into nude mice resulted in the production of adenocarcinoma which contained carcinoembryonic antigen (CEA). The nude mice tumors induced by HPA cells were interpreted as myoepithelioma in which the presence of S-100 protein and myosin were identified. On the other hand, the occurrence of squamous cell nests was frequently noted in the nude mice tumors produced by inoculation of a mixture of HSG and HPA cells. The tumor cells present in the squamous cell nests had abundant tonofilaments in the cytoplasm and were attached with tight junction and distinct desmosomes. In addition, the presence of keratin in the tumor cells composing squamous cell nests was demonstrated. When the mixture of HSG and HPA cells treated with polyethylene glycol (PEG) was transplanted into the nude mice, the tumors produced consisted almost entirely of areas showing the histologic features of anaplastic carcinoma, and did not contain all of the specific cell markers observed in the HSG or HPA tumors. The nude mice tumors induced by PEG-treated HSG or HPA cells were interpreted as adenocarcinoma and myoepithelioma, respectively, and giant cells were occasionally observed in the tumor sections. These findings indicate that neoplastic cells showing differentiation stages other than those of the original two cells can be induced in nude mice by utilizing HSG and HPA cells.     

Rapid growth and spontaneous metastasis of human germinal tumors ectopically transplanted into scid (severe combined immunodeficiency) and scid-nudestreaker mice

Xenograft acceptance, growth and spontaneous metastasis of ectopically transplanted human germinal tumors were compared among scid mice, athymic nude mice and F2 hybrids constructed from scid and nude mice, in relation to the impairments of T and B cell functions in these mice. In scid mice which are deficient in T and B cell functions, human yolk sac tumor (YST-2) that originated from the ovary grew to enormous sizes in 100% of the animals after both subcutaneous and intraperitoneal transplantation, while only half (59.1% and 51.9%) of the subcutaneous and none of the intraperitoneal transplants were accepted in usual athymic nude mice (BALB/c-nu/nu and CD1-nu/nu). The YST-2 grew rapidly in scid mice, developing 3 to 10 times larger tumors compared to nude-streaker (AKR/J-nustr/nustr) and usual nude mice, respectively. Furthermore, ectopically transplanted tumors spontaneously metastasized to distant organs (mostly to the lung) in scid mice (but less frequently in leaky scid mice), while metastases have never been found in nude mice. Although a xenograft of human classic (typical or pure) seminoma of the testis has never been established in nude mice, it grows slowly in one-third (36.4%) of scid mice and very rapidly in all of scid-nustr (scid/scid; nustr/nustr) double mutant mice. Spontaneous metastases of xenografted seminomas were also observed in distant organs (lymph node, lung, liver, spleen, and kidney). The metastastic distribution of the two human germinal tumors in scid and scid-nustr mice mimics that found in human. These results (xenograft acceptance, growth of transplanted tumors and degree of metastatic spread) were compatible with the level of T and B cell impairments indicated by FACS analysis, as well as mitogen responses, serum IgG and morphological features of the thymus.