胃癌SGC-7901细胞株侧群细胞分选及生物学特性的初步研究

中国生物工程杂志

2011, Vol. 31

Issue (11): 38-43

研究报告

胃癌SGC-7901细胞株侧群细胞分选及生物学特性的初步研究

荣黎, 吴小翎

重庆医科大学附属第二医院重庆 400016

Isolation and Identification of Side Population Cells from SGC-7901 Cell Strain

RONG Li, WU Xiao-ling

Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China

全文: PDF(846 KB) HTML

摘要：

目的:分选胃癌细胞株中的侧群(side population,SP)细胞并初步研究其相关生物学特性。方法:选择人胃癌细胞株SGC-7901,以荧光染料Hoechst 33342染色,维拉帕米拮抗对照,应用流式细胞仪检测并分选出SP细胞和nonSP细胞。CCK-8法观察两组细胞体外增殖活性;体外耐药实验检测两组细胞对化疗药物5-FU的耐药存活率;无血清培养基培养观察肿瘤球形成能力;荧光定量PCR检测干细胞相关基因Musashi-1和CD44在两组细胞中的表达差异;裸鼠体内成瘤实验观察两组细胞体内成瘤能力。结果:胃癌细胞株SGC-7901中SP细胞的比例为2.8%,与nonSP细胞相比,SP细胞具有较强的体外增殖活性(P<0.05),对5-FU的耐药存活率明显高于nonSP细胞(P<0.05),在无血清培养基中能形成明显的肿瘤球,SP细胞中Musashi-1和CD44mRNA的相对表达量明显高于nonSP细胞(P<0.05),裸鼠体内成瘤实验表明,皮下注射2×10³ 个SP细胞就能形成肿瘤,而2×10⁴ 个nonSP细胞也不能形成肿瘤。结论:胃癌细胞株SGC-7901中存在数量极少的SP细胞,SP细胞具有肿瘤干细胞的相关生物学特性。

关键词： 胃癌SGC-7901细胞; 侧群细胞; Musashi-1; CD44; 胃癌干细胞

Abstract:

Objective: To separate SP cells from SGC-7901 and identify their biocharacteristics. Methods: SGC-7901 cells were sorted as SP cells and nonSP cells using flow cytometry after SGC-7901 cells were stained with Hoechst 33342. The proliferation of two group cells in vitro was detected using CCK-8; Drug resistance experiment was used to observe two group cells resistance to 5-FU; The formation of tumor spheres was observed in serum-free culture media; The mRNA expression of Musashi-1 and CD44 was detected by real-time PCR; The tumorigeness in nude mouses was observed after nude mouses had been subcutaneously injected SP cells and nonSP cells for 8 weeks. Results: The proportion of SP cells in SGC-7901 is 2.8%;compared with nonSP cells,SP cells have stronger proliferation ability and higher resistance to 5-FU,SP cells could form obvious tumor spheres and have a higher expresssion of Musashi-1 and CD44, experiment in nude mouse demonstrates that 2×10³ SP cells could form tumor after injecting into nude mouse subcutaneously while 2×10⁴ nonSP cells could not form tunor. Conclusion:There is a small proportion of SP cells in SGC-7901,SP cells have relative biocharacteristics of tumor stem cells.

Key words: SGC-7901 cells Side population cells Musashi-1 CD44 Gastric tumor stem cell

收稿日期: 2011-07-13 出版日期: 2011-11-25

ZTFLH:

Q813

通讯作者: 吴小翎 E-mail: wuxiaoling@cqmu.edu.cn

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

引用本文:

荣黎, 吴小翎. 胃癌SGC-7901细胞株侧群细胞分选及生物学特性的初步研究[J]. 中国生物工程杂志, 2011, 31(11): 38-43.

RONG Li, WU Xiao-ling. Isolation and Identification of Side Population Cells from SGC-7901 Cell Strain. China Biotechnology, 2011, 31(11): 38-43.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I11/38

[1] 何欣.胃癌干细胞研究进展.中国肿瘤临床与康复,2009,16(3):281-282. He X. Chinese Journal of Clinical Oncology and Rehabilitation, 2009,16(3):281-282.

[2] 巩超捷,夏璐.侧群细胞与消化系恶性肿瘤干细胞研究.胃肠病学与肝病学杂志,2010,19(3):196-198. Gong C J, Xia L.Journal of Gastroenterology and Hepatology, 2010,19(3):196-198.

[3] 张祎,魏素菊,韩文峰.肿瘤干细胞与消化道肿瘤的研究进展.中国老年学杂志,2010,30(12):1757-1759. Zhang W, Wei S J, Han W F. Journal of Chinese Gerontics, 2010,30(12):1757-1759.

[4] 张媛媛.肿瘤干细胞与消化系统肿瘤.胃肠病学与肝病学杂志,2010,18(4):312-315. Zhang Y Y. Journal of Gastroenterology and Hepatology, 2010,18(4):312-315.

[5] Bonnet D, Dick J E. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell.Nat Med,1997,3(7): 730-737.

[6] Al-Hajj M, Wicha M S, Benito-Hernandez A,et al. Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA, 2003,100(7): 3983-3988.

[7] Singh S K, Hawkins C, Clarke I D,et al. Identification of human brain tumour initiating cells. Nature, 2004,432(7015):396-401.

[8] OBrien C A, Pollett A, Gallinger S,et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice.Nature,2007,445(7123):106-110.

[9] Yang Z F, Ho D W, Ng M N,et al. Significance of CD90+cancer stem cells in human liver cancer.Cancer Cell,2008,13(2):153-166.

[10] Goodell M A, Brose K, Paradis G, et al.Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J Exp Med, 1996, 183(4): 1797-1806.

[11] Patrawala L,Calhoun T,Sclmeider-Broussard R,et al.Side population is enriched in tumorigenic,stem-like cancer cells,whereas ABCG2+ and ABCG2-cancer cells are similarly tumorigenic. Cancer Res,2005,65(14):6207-6219.

[12] Piccirillo S G, Binda E, Fiocoo R,et al.Brain cancer stem cells. Mol Med, 2009,87(11):1087-1095.

[13] Haraguchi N, Inoue H, Tanaka F, et al.Cancer stem cells in human gastroiniestinal cancers.Hum Cell,2006,19(1):24-29.

[14] Haraguchi N,Utsunomiya T, Inoue H,et al.Characterization of a Side Population of Cancer Cells from Human Gastrointestinal System.Stem Cells,2006,24(3):506-513.

[15] Ho M M,Ng A V,Lam S,et al.Side population in human lung cancer cell lines and tumor is enriched with stem-like cancer cells.Cancer Res,2007,67(10):4827-4833.

[16] Chiba T, Kita K,Zheng Y W,et al.Side population purified from hepatocellular carcinoma cells harbors cancer stem cell-like properties.Hepatology,2006,44(1):240-251.

[17] Forbes S J,Alison M R,Side population(SP)cells:taking center stage in regeneration and liver cancer? Hepatology,2006,44(1):23-26.

[18] Fukuda K, Saikawa Y,Ohashi M, et al.Tumor initiating potential of side population cells in human gastric cancer.International journal of oncology,2009,34(5):1201-1207.

[19] AI-Haij M, Wicha M S,Benito-Hernandes A,et al.Prospective ideniification of tumorigenic breast caner cells.Proc Natl Acad Sci USA,2003,100(7):3983-3988.

[20] Collins A T,Berry P A,Hyde C,et al.Prospective identification of tumorigenic prostate cancer stem cells.Cancer Res,2005,65(23):10946-10951.

[21] Li C, Heidt D G,Dalerba P, et al.Identification of pancreatic cancer stem cells.Cancer Res,2007,67(3):1030-1037.

[22] Prince M E,Sivanadan R, Kaczorowski A, et al.Identification of subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma.Proc Natl Acad Sci USA, 2007,104(3):973-978.

[23] Takaishi S, Okumura T, Tu S,et al.Identification of Gastric Cancer Stem Cells Using the Cell Surface Marker CD44. Stem Cells,2009,27(5): 1006-1020.

[24] Nagata H, Akiba Y, Suzuki H, et al.Expression of Musashi-1 in the rat stomach and changes during mucosal injury and restitution.FEBS lett,2006,580(1):27-33.

[25] Akasaka Y, Saikawa Y, Fujita K, et al.Expression of a candidate marker for progenitor cells,Musashi-1,in the proliferative regions of human antrum and its decrease expression in intestinal metaplasia. Histopathology,2005,47(4):348-356.

[1]	申斓, 周爱东, 吴小芹. 植物细胞培养生物反应器的种类特点及展望[J]. 中国生物工程杂志, 2015, 35(8): 109-115.
[2]	李智力, 易小萍, 储炬, 庄英萍, 张嗣良. PK-15细胞微载体悬浮培养生产猪细小病毒的工艺研究[J]. 中国生物工程杂志, 2015, 35(7): 62-67.
[3]	张银川, 刘萌萌, 张雅婷, 桂芳, 张爱华, 闭兰, 潘勇兵. 表达全人源抗人IgE单抗的细胞株构建及筛选[J]. 中国生物工程杂志, 2015, 35(3): 66-74.
[4]	肖虎松, 沈晓涛, 郑馨, 李艳梅, 蔡冬青. 基于c-Kit⁺显微形态标记及流式分选分离心脏Telocytes[J]. 中国生物工程杂志, 2015, 35(1): 46-53.
[5]	吕志伟, 吴文平, 毕丽伟, 江星星, 王雅英. 建立金线莲原球茎悬浮体系生产次生代谢产物[J]. 中国生物工程杂志, 2012, 32(05): 43-50.
[6]	傅俊英. 干细胞领域研究、开发及市场的全球态势分析[J]. 中国生物工程杂志, 2011, 31(9): 132-139.
[7]	卜晓倩, 张瑞, 申慧琴, 罗静, 刘燕, 张路英, 刘春亮, 王琦. 吲哚胺2,3-双加氧酶基因转染对肝癌细胞凋亡的影响及相关机制研究[J]. 中国生物工程杂志, 2011, 31(5): 22-27.
[8]	杨晖, 郭琪, 赵长崎, 周剑平. 珍稀药用植物桃儿七细胞工程及内生菌研究进展[J]. 中国生物工程杂志, 2010, 30(11): 94-99.

Viewed

Full text

Abstract

Cited

Shared

Discussed