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1.
Cell lines are very useful for clinical and basic research. The establishment of uterine malignant tumor cell lines with unusual histology is especially important. We describe the establishment and characterization of a new human endometrial stromal sarcoma cell line of the uterus. The cell line OMC-9 was established from a tumor mass in the uterine body of a 55-year-old woman. Characteristics of the cell line studied include morphology, chromosome analysis, heterotransplantation, tumor markers and chemosensitivity. This cell line has grown well for 196 months and has been subcultured more than 50 times. Monolayer cultured cells are polygonal in shape, appear to be spindle-shaped or multipolar and have a tendency to pile up without contact inhibition. The cells exhibit a human karyotype with a modal chromosomal number in the diploid range. The cells were able to be transplanted into the subcutis of nude mice and produced tumors resembling the original tumor. OMC-9 cells produced tissue polypeptide antigen. Both CD10, a sensitive and diagnostically useful marker of endometrial stromal neoplasms, and vimentin were identified immunohistochemically in the original tumor and the heterotransplanted tumor. The cells were sensitive to actinomycin D, doxorubicin, carboplatin, cisplatin and etoposide, drugs used commonly in the treatment of gynecologic cancer. Only three reports of uterine endometrial stromal sarcoma cell lines have thus far been reported in the literature. OMC-9 is the first endometrial stromal sarcoma cell line in which CD10 expression and chemosensitivity have been identified. 相似文献
2.
Cancer invasion and metastasis, involving a variety of pathological processes andcytophysiological changes,contribute to the high mortality of lung cancer.The type 1 insulin-like growthfactor receptor (IGF-1R),associated with cancer progression and invasion,is a potential anti-invasion andanti-metastasis target in lung cancer.To inhibit the invasive properties of lung cancer cells,we successfullydown-regulated IGF-1R gene expression in A549 human lung cancer cells by small interfering RNA (siRNA)technology,and evaluated its effects on invasion-related gene expression,tumor cell in vitro invasion,andmetastasis in xenograft nude mice.A549 cells transfected with a plasmid expressing hairpin siRNA forIGF-1R showed a significantly decreased IGF-1R expression at the mRNA level as well as the proteinlevel.In biological assays,transfected A549 cells showed a significant reduction of cell-matrix adhesion,migration and invasion.Consistent with these results,we found that down-regulation of IGR-1Rconcomitantly accompanied by a large reduction in invasion-related gene expressions,including MMP-2,MMP-9,u-PA,and IGF-1R specific downstream p-Akt.Direct tail vein injections of plasmid expressinghairpin siRNA for IGF- 1R significantly inhibited the formation of lung metastases in nude mice.Our resultsshowed the therapeutic potential of siRNA as a method for gene therapy in inhibiting lung cancer invasionand metastasis. 相似文献
3.
Objectives: The key to fertility in adult males is production of mature spermatogenic cells. Spermatogonial stem cells (SSC) have the dual capacity of self-renewal and of differentiation into mature sperm. SSC transplantation may provide potential treatment for specific male infertilities. However, until now, there has been no evidence of offspring produced by transplantation of adult SSC line cells in humans or other mammals.
Materials and methods: A new line of SSCs from adult C57BL/6 mouse was established by using magnetic-activated cell sorting. The cell line was characterized by immunocytochemistry, karyotype analysis and telomeric repeat amplification protocol (TRAP) telomerase activity assay. Spermatogenic function was examined by allograft into germ cell-ablated recipient mice.
Results: For more than 14 months with more than 65 maintenance passages, the cell line showed a normal karyotype (40, XY) and high telomerase activity. It represented a Thy-1+, Oct4+, SSEA-1-, c-kit- (99 ± 1%) cell subpopulation. We cryopreserved these SSCs and successfully produced normal offspring after transplanting them into testes of busulphan-sterilized mice.
Conclusions: We established and long-term maintained an adult SSC line with normal spermatogenic function, without the need of genetic modification; thus, this study provides a model system for basic research and clinical application. 相似文献
Materials and methods: A new line of SSCs from adult C57BL/6 mouse was established by using magnetic-activated cell sorting. The cell line was characterized by immunocytochemistry, karyotype analysis and telomeric repeat amplification protocol (TRAP) telomerase activity assay. Spermatogenic function was examined by allograft into germ cell-ablated recipient mice.
Results: For more than 14 months with more than 65 maintenance passages, the cell line showed a normal karyotype (40, XY) and high telomerase activity. It represented a Thy-1+, Oct4+, SSEA-1-, c-kit- (99 ± 1%) cell subpopulation. We cryopreserved these SSCs and successfully produced normal offspring after transplanting them into testes of busulphan-sterilized mice.
Conclusions: We established and long-term maintained an adult SSC line with normal spermatogenic function, without the need of genetic modification; thus, this study provides a model system for basic research and clinical application. 相似文献
4.
The cell line designated HHUABM was established from the metastatic region (left Bartholin gland) of human endometrial adenocarcinoma. The cell line grew well, multilayering rapidly without contact inhibition, and 72 serial passages were successively done within 25 months. The cultured cells of HHUABM line were round and spindle in shape, and showed a pavement-like arrangement. The distribution of chromosome number varied narrowly at the diploid range, and the modal chromosome number was 46. The 90% of metaphase cells showed normal karyotype. The HHUABM cells were transplanted easily into the subcutis of BALB/c nude mice and produced poorly differentiated adenocarcinoma resembling the original tumor. The conditioned medium promoted the proliferation of CPAE (endothelial cells). The estradiol-17 beta and progesterone receptors were not detected. 相似文献
5.
Graeser R Vrignaud P Esser N Umber S Zirrgiebel U Chiron M Schaechtele C Kubbutat MH 《Journal of biomolecular screening》2012,17(3):339-349
The insulin-like growth factor I receptor (IGF1-R) system has long been implicated in cancer and is a promising target for tumor therapy. Besides in vitro screening assays, the discovery of specific inhibitors against IGF-1R requires relevant cellular models, ideally applicable to both in vitro and in vivo studies. With this aim in mind, the authors generated an inducible cell line using the tetracycline-responsive gene expression system to mimic the effects of therapeutic inhibition of the IGF-1R both in vitro and on established tumors in vivo. Inducible overexpression of IGF-1R in murine embryonic fibroblasts was achieved and resulted in the transformation of the cells as verified by their ability to grow in soft agar and in nude mice. Continuous repression of exogenous IGF-1R expression completely prevented outgrowth of the tumors. Furthermore, induced repression of IGF-1R expression in established tumors resulted in regression of the tumors. Interestingly, however, IGF-1R-independent relapse of tumor growth was observed upon prolonged IGF-1R repression. The IGF-1R cell line generated using this approach was successfully employed to test reference small-molecule inhibitors in vitro and an IGF-1R-specific inhibitory antibody, EM164, in vivo. Besides efficacy as a read-out, phospho-AKT could be identified as a pharmacodynamic biomarker, establishing this cell line as a valuable tool for the preclinical development of IGF-1R inhibitors. 相似文献
6.
Insulin-like growth factor-1 receptor (IGF-1R) is a cell membrane receptor with tyrosine kinase activity and plays important roles in cell transformation, tumor growth, tumor invasion, and metastasis. Picropodophyllin (PPP) is a selective IGF-1R inhibitor and shows promising antitumor effects for several human cancers. However, its antitumor effects in nasopharyngeal carcinoma (NPC) remain unclear. The purpose of this study is to investigate the antitumor activity of PPP in NPC using in vitro cell culture and in vivo animal model. We found that PPP dose-dependently decreased the IGF-induced phosphorylation and activity of IGF-1R and consequently reduced the phosphorylation of Akt, one downstream target of IGF-1R. In addition, PPP inhibited NPC cell proliferation in vitro. The half maximal inhibitory concentration (IC50) of PPP for NPC cell line CNE-2 was ?1 μM at 24 h after treatment and ?0.5 μM at 48 h after treatment, respectively. Moreover, administration of PPP by intraperitoneal injection significantly suppressed the tumor growth of xenografted NPC in nude mice. Taken together, these results suggest targeting IGF-1R by PPP may represent a new strategy for treatment of NPCs with positive IGF-1R expression. 相似文献
7.
Chuanliang Xu Weidong Xu Shancheng Ren Chengyao Wu Fubo Wang Ji Lu Yi Sun Jian Shen Min Wei Zhengsheng Zhang Xu Gao Yinghao Sun 《In vitro cellular & developmental biology. Animal》2013,49(9):668-678
Muscle-invasive bladder cancer is prone to metastasis without a standard organ preference. The current cell lines used to study bladder cancer have primarily been derived from individuals in Western populations, and no human bladder cancer cell line has been established from the Chinese population. A bladder cancer cell line was derived from a female Chinese patient with muscle-invasive bladder cancer, and these cells were then xenografted into the bladders of three nude mice. Five weeks later, these mice were killed to observe local invasion and distant metastasis. The metastatic tumors were also removed and analyzed to assess the metastatic mechanism. This bladder cancer cell line, named T921, was successfully established, as evidenced by karyotype and immunohistochemistry analyses. Multi-organ metastases were observed in all three of the nude mice 5 wk after the orthotopic transfer of the cell line. In addition, epithelial–mesenchymal transition (EMT)-related genes were involved in the tumor metastases. The T921 bladder cancer cell line was successfully established, and EMT was observed to play a role in bladder cancer metastasis. 相似文献
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9.
用反转录PCR从正常人胚胎肺细胞中获得了p21基因cDNA,将其插入真核表达载体pMSCVneo,构建成重组质粒,pMS21,并将其转染至肺癌细胞株A549.通过集落形成观察到p21对肺癌细胞具有明显的抑制作用,经RNA狭缝杂交、West-ernblot分析和免疫细胞化学实验证实这是p21表达的结果.荷瘤裸鼠实验也进一步证实了p21对肺癌细胞具有明显的抑制作用.为p21的深入研究提供了基础. 相似文献
10.
A new cell line, designated UHKBR-01, was successfully established from a 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumour. DMBA was administered orally at a dose of 4 mg/ml per rat on the first day of the experiment and thereafter at weekly intervals of same dosage, until the rats have reached a weight of around 150-200 g. The tumours grew rapidly after the injection, and were transplanted into nude mice one the harvest size (2.5 x 2 x 1 mm(3)) was reached, it was transplanted onto nude mice. We have developed a cell line from a portion of the DMBA-induced carcinoma of the nude mice. The UHKBR-01 cell exhibited a slow increase in growth rate during the time of culture and was highly tumourigenic in nude mice. The cells have been grown in culture for over 40 passages. Characterization of the cell line was performed. This included morphology by light and transmission electron microscopy, karyotype, growth rate, tumour antigen expression and xenograft implantation into nude mice. These cells exhibit ultrastructural and immunohistochemical features of epithelial cells of mammary origin. The above analyses also demonstrated that UHKBR-01 cells were oestrogen- and progesterone-receptor positive, in likeness to other established breast cancer cell lines such as MDA-MB-231 and MCF-7. The cell line grows as monolayers of oval-shaped cells with large folded nuclei accompanied by a rich supply of mitochondria. This report describes the first in vitro cell line from transplantable DMBA-induced mammary carcinoma of nude mice, which presents unique characteristics that may prove to be a good experimental model for investigating breast cancer biology. 相似文献
11.
12.
Establishment and some mutational characteristics of 3T3-like near-diploid mouse cell line 总被引:4,自引:0,他引:4
A stable and nonmalignant near-diploid cell line, designated m5S, has been established by serial in vitro transfers of embryonic skin cells of ICR mouse. The cells are stable in karyotype and growth, display high sensitivity to contact inhibition of cell division, are nontumorigenic in nude mice, and the transformation and mutation may be reproducibly tested. Unlike most of the mouse cell lines, the m5S underwent a unique karyotypic change during establishment and showed high stability in a diploid range, making amenable to the study of mutagenesis. The m5S is nearly unique; it represents the second near-diploid cell line sensitive to postconfluence inhibition of cell division derived from mouse embryo cells to be reported and characterized. These cells are being successfully used for qualitative and quantitative studies of oncogenic transformation as well as mutation to drug resistance by radiation and chemicals. 相似文献
13.
V Diehl M Schaadt H Kirchner K P Hellriegel F Gudat C Fonatsch E Laskewitz R Guggenheim 《Blut》1978,36(6):331-338
Two long-term cell lines were established in vitro from the peripheral blood of a patient with plasma cell leukemia: one line with plasma cell proliferation, the other with lymphoblastoid cell proliferation (LCL). The 9-month-old plasma cell line showed the typical morphology of plasmoblasts. The cells neither had B- nor T-lymphocyte characterisitics, were EBV negative, and showed aneuploidy with various marker chromosomes, including the 14 q+ marker. The cytogenetic findings indicate a monoclonal proliferation of the plasmacells. No tumor growth in thymusless nude mice could be induced upon intracranial inoculation with these cells. In contrast, the autologous LCL, cultured after addition of exogenous EBV, showed the characteristic markers of lymphoblastoid cells, with the typical morphology of pear- and handmirror-shaped lymphoblasts, growing in clumps. They had C3- and Fc-receptors, surface-Ig, E-rosette-negativity, a diploid karyotype, and EBV dependent macromolecule synthesis. They lymphoblastoid cells produced intracranial tumors in nude mice in 8 out of 8 attempts. 相似文献
14.
Zhang DL Liu SG Yan LF Li LJ Huang GS Fang FD Xia GT He XY Gao BX Bai XH Wang W Ding PG 《Cell biology international》2001,25(10):997-1002
The carcinogenic or tumourigenic testing of seven animal kidney cell lines (F-81, CRFK, MDCK, Vero, Vero-2 cell line, MA-104 and BHK-21) established in China, were carried out in more than 700 nude mice for colony formation in soft agar and for agglutination under different density of plant lectins. Tests showed that there were correlation between cell line chromosome number variations and anchorage independence in soft agar, agglutinability under lectins and tumour-forming ability in nude mice. Since testing in vitro was more economical, simpler and faster and thus thought to be more reliable, we recommend measuring agglutinability, followed by anchorage independence or analysis of karyotype as the initial means for monitoring tumourigenicity of animal cell lines in nude mice. 相似文献
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16.
Cong Wang Liping Jiang Saiqi Wang Hongge Shi Junwei Wang Ran Wang Yongmei Li Yinhui Dou Ying Liu Guiqin Hou Yu Ke Hongmin Liu 《PloS one》2015,10(6)
Jesridonin, a small molecule obtained through the structural modification of Oridonin, has extensive antitumor activity. In this study, we evaluated both its in vitro activity in the cancer cell line EC109 and its in vivo effect on tumor xenografts in nude mice. Apoptosis induced by Jesridonin was determined using an MTT assay, Annexin-V FITC assay and Hoechest 33258 staining. Apoptosis via mitochondrial and death receptor pathways were confirmed by detecting the regulation of MDM2, p53, and Bcl-2 family members and by activation of caspase-3/-8/-9. In addition, vena caudalis injection of Jesridonin showed significant inhibition of tumor growth in the xenograft model, and Jesridonin-induced cell apoptosis in tumor tissues was determined using TUNEL. Biochemical serum analysis of alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT), total protein (TP) and albumin (ALB) indicated no obvious effects on liver function. Histopathological examination of the liver, kidney, lung, heart and spleen revealed no signs of JD-induced toxicity. Taken together, these results demonstrated that Jesridonin exhibits antitumor activity in human esophageal carcinomas EC109 cells both in vitro and in vivo and demonstrated no adverse effects on major organs in nude mice. These studies provide support for new drug development. 相似文献
17.
R. Michael Sramkoski Thomas G. Pretlow II Joseph M. Giaconia Theresa P. Pretlow Stuart Schwartz Man-Sun Sy Susan R. Marengo Johng S. Rhim Desheng Zhang James W. Jacobberger 《In vitro cellular & developmental biology. Animal》1999,35(7):403-409
Summary A cell line has been derived from a human prostatic carcinoma xenograft, CWR22R. This represents one of very few available
cell lines representative of this disease. The cell line is derived from a xenograft that was serially propagated in mice
after castration-induced regression and relapse of the parental, androgen-dependent CWR22 xenograft. Flow cytometric and cytogenetic
analysis showed that this cell line represents one hyper DNA-diploid stem line with two clonal, evolved cytogenetic sublines.
The basic karyotype is close to that of the grandparent xenograft, CWR22, and is relatively simple with 50 chromosomes. In
nude mice, the line forms tumors with morphology similar to that of the xenografts, and like the parental CWR22 and CWR22R
xenografts, this cell line expresses prostate specific antigen. Growth is weakly stimulated by dihydroxytestosterone and lysates
are immunoreactive with androgen receptor antibody by Western blot analysis. Growth is stimulated by epidermal growth factor
but is not inhibited by transforming growth factor-β1. 相似文献
18.
Chun-Fang Wang Guo Zhang Li-Jun Zhao Wen-Juan Qi Xiao-Ping Li Jian-Liu Wang Li-Hui Wei 《PloS one》2013,8(8)
Epidemiological studies have demonstrated that type 2 diabetes mellitus (T2DM) and hyperinsulinemia are associated closely with endometrial carcinoma risk, although the molecular mechanism remains unclear. Insulin receptor isoformA expression is upregulated in many cancer cells and tissues, which suggests that IR-A-mediated signaling pathways may have important implications for cancer pathogenesis. We measured the expression of insulin receptor isoforms (IR-A and IR–B in the normal endometrium tissues, the endometrial carcinoma tissues and the endometrial carcinoma cell lines. We found that the total insulin receptor (IR) and IR-A expression mRNA levels and the ratio of IR-A to total IR in endometrial carcinoma specimens were significantly higher than them in control endometrial tissue specimens(P<0.05). Further analysis indicated that the tendency was more prominently in patients with T2DM. IR-A mRNA was differentially expressed in four endometrial carcinoma cell lines (Ishikawa, KLE, RL95-2 and HEC-1-A. RL95-2 cells have a low endogenous IR-A expression, and these were used to construct a stable cell line overexpressing IR-A. We found that IR-A overexpression significantly increased cell proliferation, the proportion of cells in S phase, activation of the Akt pathway and tumorigenicity of xenografts in nude mice. In contrast, there was no significant difference in the the percentage of apoptotic cells between cells overexpressing IR-A and control cells. Moreover, levels of phosphorylated ERK1/2 protein were significantly decreased in cells overexpressing IR-A relative to controls. These findings reveal the pivotal role of IR-A in endometrial cancer carcinogenesis, and suggest that the association of elevated IR-A levels with cell proliferation and tumorigenicity may be causally linked to its effect on the proportion of cells in S phase and the activation of the Akt pathway. 相似文献
19.
Joan H. Schiller Gerard Bittner Shi-Qi Wu Lorraine Meisner 《In vitro cellular & developmental biology. Animal》1998,34(4):283-289
Summary In this study, we describe the karyotypic changes associated with the spontaneous acquisition of tumorigenicity in an immortalized
tumor bronchial cell line. Neoplastic transformation of the NL20 human bronchial epithelial cell line occurred after 3 yr
in culture, and was associated with loss of chromosome 18 together with acquisition of multiple copies of 9q21.2→34. The nontumorigenic
NL20 cell line had been established by transfection of human bronchial epithelial cells with the SV40 T antigen, and had retained
a relatively stable karyotype after the first 32 passages in vitro. However, when cells from p184 were inoculated into nude mice, a transplantable tumor was obtained that was derived from
a minor clone present in this otherwise stable line. Subsequent passaging of the NL20 cells in vitro did not yield further tumors, and the minor clone from which the tumorigenic NL20T cell line derived was no longer evident
in NL20 cells by Passage 205. Furthermore, the original tumorigenic NL20T cells lost the neoplastic phenotype after 25 passages
in vitro and reverted to the nontumorigenic karyotype observed at p189. In contrast to the loss of the tumorigenic phenotype and karyotype,
which occurred with in vitro passaging of the original tumor, when the NL20T cells were passaged in other nude mice, they continued to give rise to tumors
with sevenfold amplifications of 9q sequences and loss of chromosome 18, and cells from the secondary tumors (NL20T-A cells)
have maintained a stable karyotype and remain tumorigenic even after 64 passages in vitro. A mixture of 10% tumorigenic NL20T-A and 90% nontumorigenic NL20 cells formed tumors in athymic nude mice when cultured
in vitro on fibronectin, but not on plastic; cytogenetic analysis demonstrated that the tumors and cell cultures were composed of
tumorigenic NL20T-A cells, whereas cytogenetic analysis of cells cultured on plastic were identical to the nontumorigenic
NL20 cells. These data support the hypothesis that neoplastic transformation in our original cell line arose from in vivo selection of a small mutant clone, which had arisen in culture and was subsequently selected in vivo but was lost with in vitro culture. 相似文献
20.
Chen T Xu Y Guo H Liu Y Hu P Yang X Li X Ge S Velu SE Nadkarni DH Wang W Zhang R Wang H 《PloS one》2011,6(6):e20729
The present study was designed to determine the biological effects of novel marine alkaloid analog 7-(4-fluorobenzylamino)-1,3,4,8-tetrahydropyrrolo[4,3,2-de]quinolin-8(1H)-one (FBA-TPQ) on human ovarian cancer cells for its anti-tumor potential and the underlying mechanisms as a novel chemotherapeutic agent. Human ovarian cancer cells (A2780 and OVCAR-3), and Immortalized non-tumorigenic human Ovarian Surface Epithelial cells (IOSE-144), were exposed to FBA-TPQ for initial cytotoxicity evaluation (via MTS assay kit, Promega). The detailed in-vitro (cell level) and in-vivo (animal model) studies on the antitumor effects and possible underlying mechanisms of action of the compounds were then performed. FBA-TPQ exerted potent cytotoxicity against human ovarian cancer A2780 and OVCAR-3 cells as an effective inhibitor of cell growth and proliferation, while exerting lesser effects on non-tumorigenic IOSE-144 cells. Further study in the more sensitive OVCAR-3 cell line showed that it could potently induce cell apoptosis (Annexin V-FITC assay), G2/M cell cycle arrest (PI staining analysis) and also dose-dependently inhibit OVCAR-3 xenograft tumors' growth on female athymic nude mice (BALB/c, nu/nu). Mechanistic studies (both in vitro and in vivo) revealed that FBA-TPQ might exert its activity through Reactive Oxygen Species (ROS)-associated activation of the death receptor, p53-MDM2, and PI3K-Akt pathways in OVCAR-3 cells, which is in accordance with in vitro microarray (Human genome microarrays, Agilent) data analysis (GEO accession number: GSE25317). In conclusion, FBA-TPQ exhibits significant anticancer activity against ovarian cancer cells, with minimal toxicity to non-tumorigenic human IOSE-144 cells, indicating that it may be a potential therapeutic agent for ovarian cancer. 相似文献