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目的:本文通过分离提取无小RNA(miRNA)的外来体(exosome)刺激树突细胞/细胞因子活化杀伤细胞(DC/CIKs),激活其对于胰腺癌细胞的免疫杀伤作用。创新点:无miRNA的exosome超速离心裂解产物可以通过激活DC/CIKs细胞增强其对肿瘤细胞的杀伤作用。方法:通过收集PANC-1细胞的上清并超速离心提取其中的exosome。提取的DC细胞分别通过脂多糖、肿瘤来源exosome及无miRNA的exosome刺激后,与CIK细胞共培养。通过计算增值与杀伤效率,肿瘤坏死因子-α(TNF-α)及穿孔素的分泌,比较各组间CIK细胞对胰腺癌细胞的杀伤作用。结论:经无miRNA的exosome刺激后的CIK细胞比其他两组表现出更高的杀伤效应。实验结果表明无mi RNA的exosome蛋白在DC/CIKs细胞的胰腺癌治疗中是有相当前景的激动剂。
OBJECTIVE: In this study, dendritic cells / cytokine activated killer cells (DCs / CIKs) were stimulated by exosomes extracted from small RNA (miRNA) -based cells to activate their immunosuppressive effect on pancreatic cancer cells. Innovative point: miRNA-free exosomes from ultracentrifugation lysates can enhance their killing of tumor cells by activating DC / CIKs cells. Methods: The exosomes were extracted by collecting the supernatant of PANC-1 cells and ultracentrifuging. The extracted DCs were co-cultured with CIK cells stimulated by lipopolysaccharide, tumor-derived exosomes and miRNA-free exosomes, respectively. The killing effect of CIK cells on pancreatic cancer cells was compared by calculating the value-added and killing efficiency, the secretion of tumor necrosis factor-α (TNF-α) and perforin. Conclusion: CIK cells stimulated by exosomes without miRNA show higher killing effect than the other two groups. The experimental results show that exosome protein without mi RNA is a promising agonist in the treatment of pancreatic cancer in DC / CIKs cells.