噬菌体特异性结合肽对乳腺癌干细胞生物学行为的影响  

Effects of phage specific peptide on biological behavior of breast cancer stem cells

作　　者：亓春玲[1] 刘飞[2] 赵书平[1] 孔冕[3] 刘婷婷[2] 李宝江[2] 孟潘庆[2] 

机构地区：[1]泰山医学院附属泰安医院(泰安市中心医院)临床检验中心,山东泰安271000 [2]泰山医学院(泰安市中心医院)附属泰安医院肿瘤诊疗中心,山东泰安271000 [3]济宁市第一人民医院乳甲外科,山东省273100

出　　处：《中华临床医师杂志（电子版）》2016年第9期92-97,共6页Chinese Journal of Clinicians(Electronic Edition)

基　　金：山东省自然科学基金(ZR2014HL076)

摘　　要：目的检测GY-1(GYSASRSTIPGK)与乳腺癌干细胞的亲和性及特异性,探讨其对乳腺癌干细胞体外生物学行为的影响,评估GY-1作为乳腺癌干细胞靶向治疗载体的临床价值。方法根据前期噬菌体肽库筛选实验获得的十二肽GY-1(GYSASRSTIPGK)序列,人工合成GY-1,同时合成随机氨基酸序列十二肽GY-2(GSAYITRSGPSK)作阴性对照,采用酶联免疫吸附试验(ELISA)及细胞免疫荧光法分别鉴定两者与乳腺癌干细胞231-SC(分离自MDA-MB-231细胞株)的亲和性及特异性;通过生长曲线及侵袭迁移实验检测十二肽GY-1对乳腺癌干细胞231-SC恶性生物学行为的影响。结果 ELISA结果显示,GY-1组231-SC吸光度为0.501±0.009,高于MDA-MB-231细胞(0.147±0.038)和hs578bst细胞(0.152±0.036),F=262.25,P<0.001;GY-2组中三组细胞的吸光度分别为0.148±0.003、0.146±0.005和0.149±0.001,F=0.63,P>0.05。细胞免疫荧光法显示GY-1对231-SC有高度亲和特异性,生长曲线结果显示,GY-1组231-SC细胞增殖能力显著低于GY-2组和PBS空白组,差异有统计学意义(P<0.05)。侵袭实验中GY-1组231-SC细胞透过Transwell上室的细胞数为(22±2)个,低于GY-2对照组[(41±4)个]和空白对照组[(43±6)个],F=9.03,P=0.003。迁移实验中GY-1组231-SC细胞透过Transwell上室的细胞数为(66±7)个,低于GY-2对照组[(134±11)个]和空白对照组[(141±14)个],F=14.59,P<0.001。GY-2对照组和空白组差异比较均无统计学意义(P=0.78和P=0.66)。结论十二肽GY-1对靶细胞231-SC具有高度亲和特异性,并且能够抑制231-SC的生长、侵袭迁移,有可能成为乳腺癌干细胞靶向治疗的理想载体。Objective To detect the affinity and specificity of GY-1（GYSASRSTIPGK） to breast cancer stem cells, to study its influence on the biological behavior of breast cancer stem cells in vitro and evaluate the clinical value of GY-1used for carrier targeted therapy for breast cancer stem cells. Methods GY-1 was synthesized according to GYSASRSTIPGK obtained from phage 12 peptide library using breast cancer stem cell as target cell. At the same time, random amino acid sequence peptide GY-2（GSAYITRSGPSK） was synthesized and used for negative control. The affinity and specificity of GY-1 bonding to 231-SC originated from MDA-MB-231 breast cancer cell lines were identified by ELISA and immunofluorescence. The cell proliferation was tested by growth curve, the invasion and migration abilities of 231-SC were tested. Results ELISA showed that absorbance value of 231-SC in GY-1 group（0.501±0.009） was higher than those of MDA-MB-231 cells（0.147±0.038） and hs578 bst cells（0.152±0.036）, F=262.25, P〈0.001; absorbance value of three kinds of cells in GY-2 group were 0.148±0.003, 0.146±0.005, and 0.149±0.001 respectively, F=0.63, P〈0.05. Cell immunofluorescence showed that GY-1 had high affinity and specificity to 231-SC, according to growth curve results, proliferation ability of 231-SC cells in GY-1 group was significantly lower than those in GY-2 group and PBS blank group（P〈0.05）. The invasion and migration assay in transwell systems showed that the number of cells that passed the transwell membrane in GY-1 group was 22±2 and 66±7 respectively, compared with（41±4, 134±11） in GY-2 group and（43±6, 141±14） in blank control group, F=9.03, P=0.003 and F=14.59, P〈0.001. There was no significant difference between GY-2 group and blank control group respectively（P=0.78, P=0.66）. Conclusion 12 peptide GY-1 had the ability of restraining malignant biological behavior of 231-SC, it was likely to be used for the ideal carrier targeted therapy for breast cancer stem cells.

关 键 词：噬菌体特异结合多肽 GY-1(GYSASRSTIPGK) 乳腺癌干细胞 生物学行为 靶向治疗 

分 类 号：R737.9[医药卫生—肿瘤]