脂肪干细胞来源的外泌体对三阴乳腺癌细胞生物学行为的影响

Effects of ADSCs-derived exosomes on biological behavior of triple-negative breast cancer cells

目的 探讨脂肪干细胞(ADSCs)来源的外泌体对MDA-MB-231三阴乳腺癌细胞增殖、迁移及化疗耐药性的影响.方法 从吸脂患者的脂肪组织中分离培养ADSCs,流式细胞术检测表面标志物并鉴定其成脂和成骨诱导分化能力.收集ADSCs细胞培养上清,采用超滤ExoQuick-TCTM试剂盒提取外泌体并于透射电镜下鉴定其形态.采用浓度0、400μg/ml的外泌体处理MDA-MB-231细胞,CCK-8法测定细胞增殖情况,细胞划痕实验和Transwell迁移实验检测细胞迁移能力.采用CCK-8细胞毒性实验研究其对MDA-MB-231细胞阿霉素耐药性的影响.结果 CCK-8细胞增殖实验中经外泌体处理的MDA-MB-231细胞24、48 h增殖率分别为(98.24±19.24)％、(98.15±8.05)％,差异无统计学意义(P>0.05).细胞划痕实验中,6、16、24 h显微镜下实验组细胞相对迁移距离均明显增加,差异有统计学意义(P<0.01).Transwell迁移实验显示实验组和对照组穿透膜的细胞数分别为78.0±8.8和65.3±6.2,差异有统计学意义(P<0.05).0.2μg/ml阿霉素处理MDA-MB-231细胞48 h后,实验组与对照组细胞存活率为分别为(68.99±6.81)％和(61.64±3.17)％,差异无统计学意义(P>0.05).结论 脂肪干细胞来源的外泌体对MDA-MB-231细胞的增殖无明显影响,但可显著促进MDA-MB-231细胞的迁移,而对降低MDA-MB-231细胞对阿霉素的敏感性并不明显.

Objective To investigate the influence of ADSCs-derived exosomes on the proliferation, migration and chemosensitivity of MDA-MB-231 breast cancer cells.Methods ADSCs were isolated by enzyme digestion from adipose tissue of patients undergoing liposuction and theirmorphologic characters were observed using inverted microscope . Surface markers of ADSCs were determined by flowcytometry .ADSCs were detected for their ability to undergo osteogenic and adipogenic differentiation in vitro .Collect culture supernatant of ADSCs and isolate exosomes with ExoQuick-TC kit. Morphology of purified exosomes was observed with transmission electron microscopy .MDA-MB-231 cells were treated with exosomes (0, 400 μg/ml),and the cell proliferation was measured using CCK-8 assay. Wound healing assay and transwell assay were used to investigate the effect of ADSCs -derived exosomes on the migration of MDA-MB-231 cells.The chemosensitivity of MDA-MB-231 cells to Adriamycin under the influence of ADSCs-derived exosomes was measured using CCK-8 assay.Results After treatment with 400 μg/ml exosomes for 24 h and 48 h,the cell proliferation rate was (98.24 ±19.24)％ and(98.15 ± 8.05)％that of the control group, respectively,with no statistically significant difference (P>0.05).In the wound healing assay , after incubation with 400 μg/ml exosomes for 6 h, 16 h, 24 h, the relative migration distances in the experimental group increased significantly (P<0.01).Transwell migration assay showed that the numbers of penetrated cells across the membrane in the experimental group and control group were (78.0 ±8.8) and (65.3 ±6.2), respectively(P<0.05).After treatment with 0.02 μg/ml Adriamycin for 48 h,the cell viablity of MDA-MB-231 cells in the experimental group and control group was (68.99 ±6.81)％ and (61.64 ±3.17)％ respectively, with no statistically significant difference (P>0.05).Conclusions ADSC-derived exosomes had no significant influence on the proliferation of MDA-MB-231 cells,however the migration of MDA-MB-231 cells was significantly promoted .Chemosensitivity of MDA-MB-231 cells to Adriamycin was slightly reduced , but not significantly .