期刊文献+

基于耐药特性联合单细胞培养识别肝癌干细胞克隆亚群 被引量:1

Identification of liver cancer stem cells-containing cell subgroups based on resistance characteristics and unicellular culture
在线阅读 免费下载
分享 导出
摘要 目的:基于癌干细胞的耐药特性联合单细胞培养方法识别肝癌干细胞所在的克隆亚群.方法:以BEL-7404细胞系为研究对象,裸鼠体内移植瘤形成实验,并予阿霉素(adriamycin,ADM)8 mg/kg干预,待肿瘤直径为1.5 cm时取移植瘤细胞做原代培养,并在裸鼠体内连续传代4代,并将第4代移植瘤细胞命名为"BEL-7404-ADM-P4",根据成瘤时间检测肝癌干细胞富集情况.联合单细胞培养,取BEL-7404及BEL-7404-ADM-P4所形成的克隆根据克隆形态分为全克隆、部分克隆、旁克隆,检测BEL-7404及BEL-7404-ADM-P4单克隆中全克隆、部分克隆、旁克隆的癌干特性,即增殖能力、克隆形成率及悬浮球形成率;取其全克隆、部分克隆、旁克隆做hoechst33342染色,并在共聚焦显微镜下观察其染色情况;根据BEL-7404及BEL-7404-ADM-P4单克隆中全克隆、部分克隆、旁克隆的增殖能力、克隆形成率及悬浮球形成率以及hoechst33342染色情况识别癌干细胞所在的克隆亚群.结果:裸鼠体内低剂量阿霉素干预,在体内连续成瘤传代过程中,裸鼠皮下移植瘤成瘤率均为100%,且成瘤时间从第一代到第四代均有所缩短,以此达到了肝癌干细胞的初步富集;增殖能力:全克隆增殖速度最快,细胞量最大,部分克隆次之,旁克隆增殖速度最慢,并于第10天开始出现细胞皱缩死亡;克隆形成率:BEL-7404-ADM-P4-H高于BEL-7404及BEL-7404-ADM-P4-M,差异有统计学意义(P〈0.05);悬浮球形成率:仅BEL-7404-H及BEL-7404-ADM-P4-H可形成悬浮球,其余细胞系不形成悬浮球,而BEL-7404-H及BEL-7404-ADM-P4-H悬浮球形成率比较,无统计学意义(P〉0.05);BEL-7404及BEL-7404-ADM-P4单克隆中全克隆、部分克隆、旁克隆hoechst33342染色情况:BEL-7404及BEL-7404-A D M-P4单克隆中全克隆都有极少数不染和低染的细胞存在,但部分克隆、旁克隆全染,且BEL-7404单克隆中全克隆、部分克隆、旁克隆hoechst33342荧光强度均强� AIM: To identify liver cancer stem cells-contain- ing cell subgroups based on resistance character- istics and unicellular culture.ed subcutaneously in nude mice to induce tumor formation, and adriamycin (8 mg/kg) interven- tion was given. When the tumor diameter was 1.5 cm, tumor tissues were collected for primary culture. The cells were inoculated subcutane- ously in nude mice again. After four consecutive generations in nude mice, the tumor cells were named "BEL-7404-ADM-P4". Based on the tu- mor forming time, the enrichment of liver cancer stem cells was tested. Using unicellular culture, clones from BEL-7404 and BEL-7404-ADM-P4 cells were divided into holoclones, meroclones and pareclones to test the cancer stem cell char- acteristics (renewal ability, clone formation and sphere formation). Holoclones, meroclones and pareclones were stained with hoechst33342 and observed under a confocal microscope. Based on the renewal ability, clone formation rate and sphere formation rate and hoechst33342 stain- ing properties, cancer stem cells subpopulations were identified. RESULTS: The tumor formation rate was 100%, and tumor formation time was shortened from the first generation to the fourth generation, which suggested the preliminary enrichment of liver cancer stem cells. Holoclones showed the fastest growth and the largest cell volume, fol- lowed by meroclones and pareclones. On the 10th day, pareclones started to shrivel and die. The clone formation rate of BEL-7404-ADM- P4-H cells was significantly higher than those of BEL-7404 and BEL-7404-ADM-P4-M cells (P 〈 0.05). Only BEL-7404-H and BEL-7404-ADM- P4-H cells could form spheres, and there was no significant difference in sphere formation rate between BEL-7404-H and BEL-7404-ADM-P4-H cells. In BEL-7404 and BEL-7404-ADM-P4 mono- clones and holoclones, there were a small num- ber of lowly hoechst33342 stained or non-stained cells, but cells in meroclones and pareclone cells were all stained. The hoechst33342 fluorescence intensity of BEL-740
作者 陈娟 区泳芳 蔡捷 陶璐 陈相宜 邝晓聪 Juan Chen, Yong-Fang Ou, Jie Cai, Lu Tao, Xiao-Cong Kuang, Department of Pathology and Pathophysiology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China Xiang-Yi Chen, Department of Physiology, Guangxi Medi- cal University, Nanning 530021, Guangxi Zhuang Autono- mous Region, China
出处 《世界华人消化杂志》 CAS 北大核心 2014年第3期319-326,共8页 World Chinese Journal of Digestology
基金 广西自然科学基金资助项目,Nos.桂科自0991139,2013GXNSFAAO19244 广西科学研究与技术开发计划基金资助项目,No.桂科攻0993003C-1
关键词 阿霉素干预 肝癌干细胞 单细胞培养 全克隆 部分克隆 旁克隆 Adriamycin intervention Liver cancerstem cells Unicellular culture Holoclone Nero-clone Pareclone
作者简介 陈娟,硕士,主要从事癌干细胞的研究. 通讯作者:邝晓聪,副教授,硕士生导师,530021,广西南宁市双拥路22号.厂西医科大学病理与病理生理教研室.1609800049@qq.com电话:0771—5358502
  • 相关文献

参考文献5

二级参考文献98

  • 1Clement V Sanchez P de Tribolet N Radovanovic I Ruiz i Altaba A.HEDGEHOG-GLI1 signaling regulates human glioma growth, cancer stem cell self-renewal, and tumorigenicity[J].中国神经肿瘤杂志,2007,5(2):122-122. 被引量:30
  • 2Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med, 1997, 3(7): 730. 被引量:1
  • 3Al-Hajj M, Wicha MS, Benito-Hemandez A, et al. Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA, 2003, 100(7): 3983. 被引量:1
  • 4Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors. Cancer Res, 2003, 63(18) : 5821. 被引量:1
  • 5Kim CF, Jackson EL, Woolfenden AE, et al. Identification of bron chioalveolar stem cells in normal lung and lung cancer. Cell, 2005, 121(6) : 823. 被引量:1
  • 6Xin L, Lawsonn DA, Witte ON. The Scal cell surface marker enriches for a prostate-regenerating cell subpopulation that can initiate p rostate tumorigenesis. Proc Natl Acad Sci USA, 2005, 102(19): 6942. 被引量:1
  • 7Zhang HB, Ren CP, Yang XY, et al. Identification of label-retaining cells in nasopharyngeal epithelia and nasopharyngeal carcinoma tissues. Histochem Cell Biol, 2007, 127(3) : 347. 被引量:1
  • 8Wang J, Guo LP, Chen LZ, et al. Identification of cancer stem cell like side population cells in human nasopharyngeal carcinoma cell line Cancer Res, 2007, 67(8): 3716. 被引量:1
  • 9Richardson GD, Robson CN, tang SH, et al. CD133, a novel marker for human prostatic ep ithelial stem cells. J Cell Sci, 2003, 117: 3539. 被引量:1
  • 10Zhou L, Wei X, Cheng L, et al. CD133, one of the markers of cancer stem cells in Hep-2 cell line. Laryngoscope, 2007, 117(3): 455. 被引量:1

共引文献72

同被引文献14

  • 1Pappas D, Wang K. Cellular separations: a review of new challenges in analytical chemistry[J]. Anal Chim Acta, 2007, 601 : 26-35. 被引量:1
  • 2Shalek A K, Satija R, Adiconis X, et al. Single-cell transcriptomics reveals bimodality in expression and splicing in immune cells [ J ]. Nature, 2013, 498(7453) :236-240. 被引量:1
  • 3Carlo D D, Lee L P. Dynamic Single-Cell Analysis for Quantitative Biology[J]. Anal Chem, 2006, 78: 7918-7925. 被引量:1
  • 4Tang F, Barbacioru C, Wang Y,et al. mRNASeq whole-transcriptome analysis of a single cell[ J]. Nat Methods,2009,6:377-382. 被引量:1
  • 5Schneider A, Spitkovsky D, Riess P, et al. "The Good into the Pot,the Bad into the Crop!"-A New Technology to Free Stem Cells from Feeder Cells[J]. PLoS ONE, 2008, 3(11): e3788. 被引量:1
  • 6Parkhomchuk D, Amstislavskiy V, Soldatov A, et al. Use of high throughput sequencing to observe genome dynamics at a single cell level [ J ]. Proc Natl Acad Sci, 2009, 106 : 20830-20835. 被引量:1
  • 7Guo G, Luc S, Marco E, et al. Mapping cellular hierarchy by single-cell analysis of the cell surface repertoire[J]. Cell Stem Cell. 2013, 13(4) :492-505. 被引量:1
  • 8Varadarajan N, Julg B, Yamanaka Y J, et aL A high-throughput single-cell analysis of human CD8 T cell functions reveals discordance for eytokine secretion and cytolysis [ J ] - J Clin Invest, 2011,121(11): 4322-4331. 被引量:1
  • 9Peterbauer T, Heitz J, Olbrich M, et al. Simple and versatile methods for the fabrication of arrays of live mammalian cells[ J]. Lab Chip, 2006, 6: 857-863. 被引量:1
  • 10Haupt S, Grtitzner J, Thier M C, et al. Autmnated selection and harvesting of pluripotent stem cell colonies [ J ]. Biotechnol App] Biochem, 2012, 59(2) : 77-87. 被引量:1

引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部 意见反馈