肺癌肿瘤干细胞的分选及生物学特性  

Isolation and Biological Characterization of Lung Cancer Stem Cells

作　　者：张泽峰[1] 高峰[1] 王涛[1] 王瑞[1] 郭杨[1] 张国亮[1] 曾辉[1] 刘长江[1] ZHANG Zefeng;GAO Feng;WANG Tao;WANG Rui;GUO Yang;ZHANG Guoliang;ZENG Hui;LIU Changjiang(Department of Thoracic Surgery,Fourth Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第四医院胸外科

出　　处：《中国医学科学院学报》2020年第1期24-29,共6页Acta Academiae Medicinae Sinicae

基　　金：河北省2017年度医学科学院重点课题计划~~

摘　　要：目的探讨肺癌肿瘤干细胞分选及生物学特性的研究方法。方法采用免疫磁珠分选经盐酸阿霉素(ADM)诱导的人肺腺癌SPC-A-1(SPC-A-1/ADM)细胞系中的ABCG2+和ABCG2-细胞,流式细胞术和裸鼠体内移植针对ABCG2+和ABCG2-细胞体内的成瘤率进行测定分析。结果SPC-A-1细胞中的荧光强度平均为(1.001±0.014)×102,明显低于SPC-A-1/ADM细胞的(10.257±0.023)×102(t=17.320,P=0.001)。SPC-A-1细胞ABCG2/BCRP-FITC处理组与SPC-A-1细胞对照组(t=5.269,P=0.021)和SPC-A-1细胞同型对照组(t=6.869,P=0.012)间差异有统计学意义;SPC-A-1/ADM细胞对照组与SPC-A-1/ADM细胞同型对照组间差异有统计学意义(t=8.112,P=0.015)。SPC-A-1/ADM细胞ABCG2/BCRP-FITC处理组阳性率为9.8%,是SPC-A-1细胞组的39.84倍。SPC-A-1/ADM细胞ABCG2/BCRP-FITC处理组与SPC-A-1/ADM细胞组(t=9.120,P=0.005)和SPC-A-1/ADM细胞同型组(t=8.257,P=0.006)间差异有统计学意义。B群细胞阳性率是A群细胞的684倍,差异有统计学意义(t=11.235,P=0.001),且B群细胞的荧光强度较强。接种SPC-A-1细胞、A群细胞和B群细胞的裸鼠平均成瘤体积分别为(6.96±1.82)、(6.70±2.55)和(9.17±2.41)mm3,以接种B群细胞组最高,但3组间差异无统计学意义(F=2.362,P=0.086)。接种B群细胞组的成瘤率为75.00%,明显高于SPC-A-1细胞组和A群细胞组(F=19.780,P=0.002)。结论ABCG2抗体与免疫磁珠分选法结合可以从人肺腺癌SPC-A-1/ADM细胞系内分离ABCG2+细胞,其在裸鼠体内成瘤情况良好,可用于肺癌肿瘤干细胞的分选及生物学特性研究。Objective To explore the methods of screening and biological characteristics of lung cancer stem cells.Methods We selected the ABCG2+and ABCG2-cells from SPC-A-1/adriamycin(ADM)cell line induced by ADM and analyzed the tumorigenicity of ABCG2+and ABCG2-cells in vivo by flow cytometry and transplantation in nude mice.Results The average fluorescence intensity of SPC-A-1 cells was(1.001±0.014)×102,which was significantly lower than that of SPC-A-1/ADM cells[(10.257±0.023)×102](t=17.320,P=0.001);the difference was also statistically significant between the ABCG2/BCRP-FITC treatment group and the SPC-A-1 control group(t=5.269,P=0.021)and the SPC-A-1 control group(t=6.869,P=0.012)and between the SPC-A-1/ADM cell control group and the SPC-A-1/ADM cell homotype control group(t=8.112,P=0.015).The positive rate of SPC-A-1/ADM cells treated with ABCG2/BCRP-FITC was 9.8%,39.84 times higher than that of SPC-A-1 cells;it showed significant difference between the ABCG2/BCRP-FITC group and the SPC-A-1/ADM group(t=9.120,P=0.005)and the SPC-A-1/ADM group(t=8.257,P=0.006).The positive rate of group B cells was 684 times that of group A cells,and the difference was statistically significant(t=11.235,P=0.001),and the fluorescence intensity of group B cells was strong.The average tumorigenic volume of the mice inoculated with SPC-A-1 cells,group A cells,and group B cells was(6.96±1.82),(6.70±2.55),and(9.17±2.41)mm3,respectively.Among them,group B was the highest,but there was no significant difference among these three groups(F=2.362,P=0.086).The tumorigenic rate of group B cells was 75.00%,which was significantly higher than that of SPC-A-1 cells and group A cells(F=19.780,P=0.002).Conclusion ABCG2 cells from human lung adenocarcinoma SPC-A-1/ADM cell line can be isolated by ABCG2 antibody combined with immunomagnetic beads sorting method,and the tumor formation rate in nude mice can be observed to explore the identification and biological characterization of lung cancer stem cells.

关 键 词：肺癌肿瘤干细胞 ABCG2细胞 免疫磁珠分选 生物学特性 

分 类 号：R657.44[医药卫生—外科学]