Patho-physiological Analysis of Radio-resistant Cancer Stem Cells

放射抗性癌症干细胞的病理生理学分析

• 批准号: 18590335
• 负责人: SEKINE Ichiro
• 金额: $ 2.45万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590335/
• 关键词: Cancer stem cell; SP cell; Non-SP cell; THRB; SP細胞分画; c-Ab1; NF-kB; 放射線抵抗性; セネッセンス

There is increasing evidence that cancers contain their own stem-like cells called cancer stem cells(CSCs). A small subset of cells, termed side population(SP), has been identified using flow cytometric analysis. The SP cells have the ability to exclude the DNA binding dye, Hcechst33342, and are highly enriched for stem cells in many kinds of normal tissues. Because CSCs are thought to be drug resistant, SP cells in cancers might contain CSCs. We initially examined the presence of SP cells in several human thyroid cancer cell lines. A small percentage of SP cells were found in ARO(0.25%), FRO(0.1%), NPA(0.06%), and WRO(0.02%) cells but not TPC1 cells. After sorting, the SP cells generated both SP and non-SP cells in culture. The clonogenic ability of SP cells was significantly higher than that of non-SP cells. Moreover, the SP prevalence was dependent on cell density in culture, suggesting that SP cells preferentially survived at lower cell density. Microarray experiment revealed diffe … More rential gene expression profile between SP and non-SP cells, and several genes related to sternness were up-regulated. However, non-SP population also contained cells that were tumorigenic in nude mice, and non-SP cells generated a small number of SP cells. These results suggest that cancer stem-like cells are partly, but not exclusively, enriched in SP population.Thyroid hormone receptors(THRs) widely govern cell growth, differentiation and metabolism acting in a ligand-and cofactordependent manner to modulate tissue-specific gene expression. Given a large variety of genes regulated by THRs and multiplicity of cellular processes potentially influenced by THRs, we addressed the role of THRB(thyroid hormone receptor beta) in cellular radiosensitivity. On the other hand, SP-rich cell lines showed low level expression of THRB. Wild-type and mutant THRB were overexpressed in several cell lines using an adenovirus-mediated gene delivery and their effects were examined after cell exposure to gamma-rays. Wild-type THRB decreased clonogenic survival of the cell lines with low levels of endogenous THRB, retarded their growth and synergized with radiation in decreasing proliferative potential and promoting cellular senescence. These changes were accompanied by the accumulation of p21(CDKNIA, CIP1, WAF1) and p16(CDKN2A, TNK4a)inhibitors of cyclin-dependent kinases and by the decrease of Rb(retinoblastoma protein)phosphorylation. Mutant THRB produced a radioprotective effect, attenuated radiation-induced growth inhibition and cellular senescence. The results suggest that THRB may modulate cellular radiosensitivity and stress-induced senescence.These data is suggesting that radio-resistant property of SP cell is involved in THRB expression and its function. Less

越来越多的证据表明，癌症含有自己的干细胞样细胞，称为癌症干细胞（CSC）。一个小的细胞亚群，称为侧群（SP），已被确定使用流式细胞仪分析。SP细胞具有排除DNA结合染料Hcechst 33342的能力，并且在多种正常组织中高度富集干细胞。由于CSC被认为是耐药的，因此癌症中的SP细胞可能含有CSC。我们最初研究了SP细胞在几种人甲状腺癌细胞系中的存在。ARO（0.25%）、FRO（0.1%）、NPA（0.06%）和WRO（0.02%）细胞中发现了一小部分SP细胞，但TPC 1细胞中未发现。分选后，SP细胞在培养物中产生SP和非SP细胞。SP细胞的克隆形成能力明显高于非SP细胞。此外，SP的患病率是依赖于在文化中的细胞密度，表明SP细胞优先存活在较低的细胞密度。微阵列实验显示， ...更多信息 SP细胞与非SP细胞的干细胞基因表达谱差异显著，与干细胞相关的多个基因表达上调。然而，非SP群体也包含在裸鼠中致瘤的细胞，并且非SP细胞产生少量SP细胞。这些结果表明，肿瘤干细胞样细胞中存在部分富集的SP，甲状腺激素受体（Thyroid hormone receptor，THR）广泛调控细胞的生长、分化和代谢，并以配体和辅因子依赖的方式调节组织特异性基因的表达。鉴于THR调节的多种基因以及THR可能影响的多种细胞过程，我们探讨了THRB（甲状腺激素受体β）在细胞放射敏感性中的作用。另一方面，富含SP的细胞系显示THRB的低水平表达。使用腺病毒介导的基因递送在几种细胞系中过表达野生型和突变型THRB，并在细胞暴露于γ射线后检查其效果。野生型THRB降低了具有低水平内源性THRB的细胞系的克隆形成存活，延缓了它们的生长，并与辐射协同降低增殖潜力和促进细胞衰老。这些变化伴随着细胞周期蛋白依赖性激酶的p21（CDKN1A、CIP1、WAF1）和p16（CDKN2A、TNK4a）抑制剂的积累以及Rb（视网膜母细胞瘤蛋白）磷酸化的减少。突变体THRB产生辐射防护作用，减弱辐射诱导的生长抑制和细胞衰老。结果提示，THRB可调节SP细胞的辐射敏感性和应激性衰老，提示SP细胞的辐射抗性与THRB的表达和功能有关。少

项目成果

Therapeutic Implications of MIR-17-92 Cluster in Anaplastic Thyroid Cancer Cells

MIR-17-92 簇对甲状腺未分化癌细胞的治疗意义

• 发表时间: 2007
• 作者: Takakura S;et. al.
• 通讯作者: et. al.

Expressions of sonic hedgehog, patched, smoothened and Gli-1 in human intestinal stromal tumors and their correlation with prognosis.

• DOI: 10.3748/wjg.v12.i35.5687
• 发表时间: 2006-09
• 期刊: World journal of gastroenterology
• 影响因子: 4.3
• 作者: A. Yoshizaki;T. Nakayama;S. Naito;C. Wen;I. Sekine

Characterization of side population in thyroid cancer cell lines: Cancer stem-like cells are enriched partly but not exclusively

• DOI: 10.1210/en.2006-1553
• 发表时间: 2007-04-01
• 期刊: ENDOCRINOLOGY
• 影响因子: 4.8
• 作者: Mitsutake, Norisato;Iwao, Atsuhiko;Yamashita, Shunichi
• 通讯作者: Yamashita, Shunichi

X radiation up-regulates the occurrence and the multiplicity of invasive carcinom as in the intestinal tract of Apc(min/+)mice

X辐射上调Apc(min/ )小鼠肠道中侵袭性癌的发生和多重性

• 发表时间: 2007
• 期刊: Radiat Res 168(4)
• 作者: Nakayama T;et. al.
• 通讯作者: et. al.

X radiation up-regulates the occurrence and the multiplicity of invasive carcinomas in the intestinal tract of Apc(min/+) mice.

X 辐射上调 Apc(min/ ) 小鼠肠道中浸润性癌的发生和多重性。

• 发表时间: 2007
• 期刊: Radiat Res 168(4)
• 作者: Nakayama T;et. al.
• 通讯作者: et. al.