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Circulating Microvesicles from Pancreatic Cancer Accelerate the Migration and Proliferation of PANC-1 Cells

机译:来自胰腺癌的循环微囊泡促进PANC-1细胞的迁移和增殖

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摘要

Circulating microvesicles are able to mediate long-distance cell–cell communications. It is essential to understand how microvesicles from pancreatic cancer act on other cells in the body. In this work, serum-derived microvesicles were isolated from 10 patients with locally advanced pancreatic cancer and healthy controls. Using Cell Transwell and WST-1 reagents, we found that microvesicles from pancreatic cancer accelerated migration and proliferation of PANC-1 cells. Meanwhile, the proliferation of these cancer-microvesicle-treated cells (CMTCs) was affected less by 10 μM of gemcitabine relative to healthy microvesicle-treated cells (HMTCs). Next, we optimized the filter-aided sample preparation method to increase the recovery of protein samples and then applied it to the quantification of the proteome of CMTCs and HMTCs. The peptides were labeled and analyzed by liquid chromatography–tandem mass spectrometry. In total, 4102 proteins were identified, where 35 proteins were up-regulated with 27 down-regulated in CMTCs. We verified the quantitative results of three key proteins CD44, PPP2R1A, and TP53 by Western blot. The Ingenuity Pathway Analysis revealed pathways that cancer microvesicles might participate in to promote cell migration and proliferation. These findings may provide novel clues of treatment for tumorigenesis and metastasis.
机译:循环微泡能够介导长距离细胞间的通信。必须了解胰腺癌的微泡如何作用于体内其他细胞。在这项工作中,从10例局部晚期胰腺癌和健康对照患者中分离出血清来源的微囊泡。使用Cell Transwell和WST-1试剂,我们发现来自胰腺癌的微泡可促进PANC-1细胞的迁移和增殖。同时,相对于健康的微囊处理细胞(HMTC),吉西他滨对这些癌症微囊处理细胞(CMTC)的增殖影响较小。接下来,我们优化了过滤辅助样品制备方法,以提高蛋白质样品的回收率,然后将其应用于CMTC和HMTC蛋白质组的定量。标记肽并通过液相色谱-串联质谱法进行分析。总共鉴定出4102种蛋白质,其中CMTC中有35种蛋白质被上调而有27种蛋白质被下调。我们通过蛋白质印迹验证了三种关键蛋白CD44,PPP2R1A和TP53的定量结果。机能途径分析揭示了癌症微泡可能参与促进细胞迁移和增殖的途径。这些发现可能为肿瘤发生和转移提供新的治疗线索。

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