文献类型：期刊文献

中文题名：在胃癌BGC-823细胞中侧群细胞的分选及其干细胞特性研究

英文题名：Isolation side population cells from gastric cancer cell BGC-823 and research of characterization

作者：耿嘉[1];刘越敏[1];罗慧英[1,2]

第一作者：耿嘉

机构：[1]甘肃中医药大学药学院,甘肃兰州730101;[2]甘肃省中药药理与毒理重点实验室,甘肃兰州730101

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2019

卷号：35

期号：9

起止页码：873

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：甘肃省高校科研基金资助项目(2018A-048)

语种：中文

中文关键词：胃癌BGC-823细胞株;侧群细胞;增殖;迁移;侵袭

外文关键词：gastric cancer cell line BGC-823;side population cell;proliferation;migration;invasion

摘要：目的对胃癌BGC-823细胞株中的侧群(SP)细胞进行分选,并对其干细胞特性进行研究。方法将胃癌BGC-823细胞株传代培养至呈对数生长期后,取适量细胞悬液分为2组:对照组(加入SP抑制剂——终浓度为50μmol·L^(-1)维拉帕米,分离NSP细胞)和实验组(不加维拉帕米,分离SP细胞),用流式细胞仪分选出SP细胞和非侧群(NSP)细胞,研究SP细胞的干细胞特征:用噻唑蓝(MTT)法比较2种细胞的增殖能力;用细胞划痕实验比较2种细胞的迁移能力;用Transwell小室实验比较2种细胞的侵袭能力。结果从胃癌BGC-823细胞中分选得到约2%SP细胞。孵育后第4天,NSP细胞和SP细胞的增殖水平(光密度值)分别为0. 62±0. 42,0. 93±0. 17;第7天,这2种细胞的增殖水平(光密度值)分别为0. 81±0. 21,1. 96±0. 42;实验组与对照组比较,差异均有统计学意义(均P <0. 05)。经7,14,21 d孵育后,实验组中SP细胞比例分别降为(52. 03±2. 08)%,(3. 07±0. 88)%和(2. 16±0. 15)%;而对照组中SP细胞比例在21 d内基本维持在0. 03%左右,表明SP细胞不断通过分裂进行自我更新和分化,而NSP细胞基本不具有自我更新及分化的能力。经2 d孵育后,实验组与对照组在聚碳酸酯膜上每个视野中细胞数分别为(75. 22±4. 31),(22. 15±1. 77)个,组间比较差异有统计学意义(P <0. 05),提示SP细胞具有更强的侵袭能力。结论在胃癌BGC-823细胞株中,存在一定比例的SP细胞。与NSP细胞比较,SP细胞具有更强的增殖、分化、迁移和侵袭能力,提示胃癌BGC-823细胞株中分选所得的SP细胞具有肿瘤干细胞的特性。
Objective To isolate the side population(SP)cells from the gastric cancer cell BGC-823,and study the characteristics of stem cells.Methods The gastric cancer cell line BGC-823 was subcultured to logarithmic growth.Appropriate cell suspensions were divided into two groups:control group[adding SP cell inhibitor,verapamil with final concentration of 50μmol·L-1 for isolation none SP(NSP)cells]and experimental group(without verapamil for isolation SP cells).SP cells and NSP cells were separated by flow cytometry.MTT method was used to compare the proliferation ability of the two kinds of cells.Cell scratch test was used to compare the migration ability of the two kinds of cells.Transwell chamber test was used to compare the invasion ability of the two kinds of cells.Results About 2%of SP cells existed in gastric cancer BGC-823 cells.After 4 d of incubation,the proliferation(optical density values,OD values)of NSP cells and SP cells were 0.62±0.42,0.93±0.17;after 7 d of incubation,the proliferation(OD values)of NSP cells and SP cells were 0.81±0.21,1.96±0.42;compared with NSP cells,the difference was significant(all P<0.05).After 7,14,21 d of incubation,the proportion of SP cells in experimental group respectively decreased to(52.03±2.08)%,(3.07±0.88)%and(2.16±0.15)%;while the proportion of SP cells in control group remained at about 0.03%.It indicating that SP cells can self-renew and differentiate through asymmetric division in vitro,while NSP cells do not have the ability of self-renewal and differentiation.After 2 d of incubation,number of cells in each field of vision on polycarbonate membrane in experimental group and control group were(75.22±4.31),(22.15±1.77)point,the difference between two groups was significant(P<0.05).It is suggested that SP cells have stronger invasive ability.Conclusion There are a certain proportion of SP cells in gastric cancer cell line BGC-823.Compared with NSP cells,SP cells have stronger ability of proliferation,differentiation,migration and invasion,suggesting that SP cells isolated from gastric cancer BGC-823 cell line have the characteristics of cancer stem cells.