17 Establishment and characterization of cell line TMCC-2 from clear cell carcinoma of the endometrium.
Hiroshi IwabuchiH. SakunagaJ. InowakiK OkabeY. NutaharaHidenori SatoY NegishiK AkiyaMasato SakamotoMichio SugitaY Tenjin
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You have accessJournal of UrologyKidney Cancer: Basic Research (II)1 Apr 2013298 IDENTIFICATION AND CHARACTERIZATION OF CANCER STEM CELL OF RENAL CELL CARCINOMA CELL LINES Kosuke Ueda, Sachiko Ogasawara, Jun Akiba, Masamichi Nakayama, Sakiko Sanada, Shigetaka Suekane, Masanori Noguchi, Kei Matsuoka, and Hirohisa Yano Kosuke UedaKosuke Ueda Kurume, Japan More articles by this author , Sachiko OgasawaraSachiko Ogasawara Kurume, Japan More articles by this author , Jun AkibaJun Akiba Kurume, Japan More articles by this author , Masamichi NakayamaMasamichi Nakayama Kurume, Japan More articles by this author , Sakiko SanadaSakiko Sanada Kurume, Japan More articles by this author , Shigetaka SuekaneShigetaka Suekane Kurume, Japan More articles by this author , Masanori NoguchiMasanori Noguchi Kurume, Japan More articles by this author , Kei MatsuokaKei Matsuoka Kurume, Japan More articles by this author , and Hirohisa YanoHirohisa Yano Kurume, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1682AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Evidence for existence of cancer stem cells (CSCs) has been reported in several malignant tumors. The population of CD105 is reported to be one of the CSC markers in renal cell carcinoma (RCC). However, there are a few reports examining the CSCs in human RCC. In our study, we isolated side population (SP) cells from the human RCC cell lines and systematically identified the CSC characteristics of the SP cells. Moreover, we examined the malignant potential with CSC like biological features of ALDH1-positive cells. METHODS We used two RCC cell lines derived from metastatic RCC (ACHN) and primary RCC (KRC/Y). The SP and Non-SP(NSP) cells were isolated from the two RCC cell lines by FACS Aria 2. We examined proliferative ability, drug resistance, clonoginity, sphere forming ability in serum-free medium in vitro and tumorigenicity in vivo between SP and NSP cells. We examined ALDH1 activity and sphere forming ability in ALDH1 expression. Moreover, we analyzed the expression of ALDH1 after treatment with Sorafenib or IFNa and exposing hypoxia. RESULTS The SP cell rates in ACHN and KRC/Y were 1.4% and 1.7%, respectively. SP cells of KRC/Y had higher cell proliferative ability and clonogenity in vitro. Although SP cells in KRC/Y expressed higher CD105-positive cell rate than NSP cells (SP vs NSP: 24.6% vs 4.6%), there was no difference in tumorigenicity in vivo. On the other hand, there was no significant difference in the cell growth rate and colony formation between SP and NSP cells in ACHN. However, the SP cells in ACHN had higher sphere forming ability and IFNa resistance in vitro and had higher tumor growth ability in vivo. The mRNA expression levels of stemness genes, HIF1a and VEGFA in isolated SP and Non-SP cells showed no significant difference. Whereas, the SP cells in ACHN expressed higher ALDH1-positive cells, which was widely known as one of the representative CSC markers, compared with NSP cells (SP vs NSP: 32.7% vs 14.6%). Furthermore, ALDH1-positive cell had higher sphere forming ability than ALDH1-negative cells. ALDH1-positive cells rate, especially treated with sorafenib or exposed in hypoxic condition, was much higher than in the control. CONCLUSIONS Cells with the features of CSC may be involved in SP cell fraction, especially in ALDH1-positive SP cell fraction. SP cells in ACHN not only overexpress ALDH1 activity but also have more resistance to the various conventional treatment for RCC. These findings may provide new insights for future CSC research and clinical anti-cancer therapy for RCC. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e121 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kosuke Ueda Kurume, Japan More articles by this author Sachiko Ogasawara Kurume, Japan More articles by this author Jun Akiba Kurume, Japan More articles by this author Masamichi Nakayama Kurume, Japan More articles by this author Sakiko Sanada Kurume, Japan More articles by this author Shigetaka Suekane Kurume, Japan More articles by this author Masanori Noguchi Kurume, Japan More articles by this author Kei Matsuoka Kurume, Japan More articles by this author Hirohisa Yano Kurume, Japan More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...
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You have accessJournal of UrologyKidney Cancer: Basic Research1 Apr 2011112 CHARACTERIZATION OF CANCER STEM CELL-LIKE CELLS AND STROMAL CELLS IN RENAL CELL CARCINOMA Daniel Rottke, Verena Börger, Peter Albers, Margit Fisch, Rolf Ackermann, and Rüdiger Sorg Daniel RottkeDaniel Rottke Hamburg, Germany More articles by this author , Verena BörgerVerena Börger Düsseldorf, Germany More articles by this author , Peter AlbersPeter Albers Düsseldorf, Germany More articles by this author , Margit FischMargit Fisch Hamburg, Germany More articles by this author , Rolf AckermannRolf Ackermann Düsseldorf, Germany More articles by this author , and Rüdiger SorgRüdiger Sorg Düsseldorf, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.178AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Cancer stem cells are crucial to the development and progression of tumors. Non-tumorigenic tissue stem cells such as mesenchymal stem cells (MSC)-like cells constitute components of tumor stroma and may also contribute to these processes. We have characterized the stem cell features of two cell lines with mesenchymal morphology, DH-1 and GF-1, derived from metastases of clear cell renal cell carcinomas (RCC), and one cell line, MG-1, with epithelial appearance derived from a primary papillary type 1 RCC. METHODS Cell lines were characterized by morphology, flow cytometry, cytogenetics, RT-PCR expression, their differentiation potential and by xenotransplantation into mice. RESULTS The mesenchymal cell lines lacked cytogenetic aberrations typical for RCC and did not form tumors in mice. GF-1 and DH-1 cells had an immunophenotype in common with bone marrow-derived MSC. Like MSC, both cell lines showed osteogenic differentiation. When co-cultured with MCF7 carcinoma cells, GF-1 cells but only to a lesser extent DH-1 cells or MSC induced epithelial-to-mesenchymal transition with change in morphology and down-regulation of E-cadherin in MCF7 cells. Upon co-transplantation, only GF-1 cells promoted tumor formation in the various mouse models. The epithelial cell line MG-1 had a similar immunophenotype, but in contrast to the mesenchymal cells, they were CD90-, CD133+, CD326+ and cytokeratin 8/18+ and revealed chromosomal aberrations typical for papillary RCC. They formed spheres and showed osteogenic but not adipogenic differentiation. When transplanted into mice, they formed tumors from which tumorigenic cells could be regrown. Subcloning of CD133+ and CD133- subpopulations of MG-1 cells resulted in immunophenotypically similar cells with osteogenic differentiation potential. However, tumor-formation was observed for CD133- clones only, with lower numbers of cells initiating tumor formation than in the parental line, suggesting that in papillary RCC cancer stem cells reside in the CD133- population. CONCLUSIONS Thus, the newly established epithelial RCC cell line MG-1- subclones share typical features with cancer stem cells, including immunophenotype, tumorigenicity, self-renewal and differentiation potential. In contrast, the mesenchymal cell lines DH-1 and GF-1 show characteristics of MSC and apparently are derived from RCC stroma. However, only GF-1 cells but not DH-1 cells or MSC have potent tumor stroma activity and induce epithelial-to-mesenchymal transition in vitro and promote tumor-formation by a normally non-tumorigenic RCC cell line in vivo. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e47 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Daniel Rottke Hamburg, Germany More articles by this author Verena Börger Düsseldorf, Germany More articles by this author Peter Albers Düsseldorf, Germany More articles by this author Margit Fisch Hamburg, Germany More articles by this author Rolf Ackermann Düsseldorf, Germany More articles by this author Rüdiger Sorg Düsseldorf, Germany More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...
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ヒト大腸癌細胞株LMを用い, ヌードマウスの脾注, 肝転移形成を3回および5回繰り返すことにより, 細胞株LM-H3およびLM-H5を樹立した.LM-H3およびLM-H5はLMに比べて有意な肝転移能の増強を認め, 電子顕微鏡による観察では, microvilliの増加を認めた.核DNA量および染色体数はLMに比べ, 肝高転移株において減少した.培養上澄中のSLA産生量は, LMに比べ, LM-H3, LM-H5で, 3倍および4.5倍の産生量を認め, Flow cytometryによる細胞表面のSLAの発現の検討でも, M.F.I.が, 102.3±43.5,126.1±28.4,144.8±23.4と有意な増加を認めた.また, 血管内皮細胞への接着性もLMに比べ, 肝高転移株において有意に増加した.以上の結果から, このように肝転移形成を繰り返すことにより, LM細胞の肝転移能は増強し, また肝高転移株は親株とは異なったさまざまな特性を有すると考えられた.
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Clear cell carcinoma (CCC) is a rare subtype of ovarian cancer resistant to standard platinum chemotherapy, which leads to a poor prognosis for patients with CCC. Kinases are targets for anticancer drugs; few studies have profiled kinase activity to identify kinase inhibitors as novel anticancer drugs. In this study, we aimed to identify novel anticancer drugs for the treatment of CCC with comprehensive kinase activity assay and drug screening. Using ascites from a 51-year old patient, we established and characterized the NCC-cOV1-C1 cell line. We screened the antiproliferative effects of 152 small anticancer compounds and conducted comprehensive kinase activity assays with the PamStation12 platform. The NCC-cOV1-C1 cells harbor copy number variation of HFN1β amplification, and exhibit constant growth, spheroid formation, and invasion capability. NCC-cOV1-C1 cells responded remarkably to idarubicin HCl and vorinostat. The kinase activity assay revealed that SRC and EGFR were highly activated in NCC-cOV1-C1 cells; the SRC inhibitor dasatinib and the EGFR inhibitor lapatinib exhibited antiproliferative effects and down-regulation of downstream signaling. The NCC-cOV1-C1 cell line will be a useful tool for basic and preclinical study of CCC, and the clinical utility of idarubicin HCl, vorinostat, dasatinib, lapatinib is worthy of further investigation.
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Carcinoma Cell
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