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用小狮子鱼(Trachinus Vipera)纯化毒液处理后对结肠癌细胞迁移的抑制作用

Inhibition of Colon Carcinoma Cell Migration Following Treatment with Purified Venom from Lesser Weever Fish (Trachinus Vipera).

作者信息

Fezai Myriam, Slaymi Chaker, Ben-Attia Mossadok, Kroemer Guido, Lang Florian, Jemaà Mohamed

机构信息

Laboratory of Biomonitoring of the Environment (LR01/ES14), Faculty of Sciences of Bizerte, Bizerte, Tunisia.

University of Carthage, Carthage, Tunisia.

出版信息

Cell Physiol Biochem. 2017;41(6):2279-2288. doi: 10.1159/000475646. Epub 2017 Apr 26.

Abstract

BACKGROUND

Injury by the sting of Lesser weever fish (Trachinus vipera) may lead to severe pain, edema or tissue necrosis. Cellular effects of the venom are still incompletely understood. Previous observations revealed that purified Lesser weever fish venom (LWFV) induces suicidal death of erythrocytes and HCT116 human colon carcinoma cells. The present study addressed the effect of the venom on colon carcinoma cell toxicity, shape and migration both in p53+/+ and/or p53-/- conditions.

METHODS

Cells were exposed to medium without or with 500 µg/ ml LWFV. Cell shape, cell area and circularity were visualized and quantified by fluorescence microscopy. Cell volume, granularity and cells toxicity were assessed via the apoptotic parameters dissipation of mitochondrial inner transmembrane potential, phosphatidylserine surface exposure and cell membrane permeabilization were measured utilizing flow cytometry. Cell migration was evaluated using wound healing assay and two-dimensional migration assay.

RESULTS

LWFV treatment was followed by a marked change of cell shape and size, significant decrease of cell area and circularity, significant impairment of cell migration, as well as induction of apoptosis after long exposition.

CONCLUSIONS

LWFV exposure leads to cell shrinkage, increased granularity, apoptosis and impairment of cell migration, effects presumably contributing to LWFV-induced tissue injury.

摘要

背景

小毒鲉(Trachinus vipera)蜇伤可导致剧痛、水肿或组织坏死。毒液的细胞效应仍未完全了解。先前的观察表明,纯化的小毒鲉毒液(LWFV)可诱导红细胞和HCT116人结肠癌细胞自杀性死亡。本研究探讨了该毒液在p53+/+和/或p53 -/-条件下对结肠癌细胞毒性、形态和迁移的影响。

方法

将细胞暴露于不含或含有500μg/ml LWFV的培养基中。通过荧光显微镜观察并量化细胞形态、细胞面积和圆形度。通过凋亡参数评估细胞体积、颗粒度和细胞毒性,利用流式细胞术测量线粒体内膜电位消散、磷脂酰丝氨酸表面暴露和细胞膜通透性。使用伤口愈合试验和二维迁移试验评估细胞迁移。

结果

LWFV处理后,细胞形态和大小发生显著变化,细胞面积和圆形度显著降低,细胞迁移显著受损,长期暴露后诱导凋亡。

结论

暴露于LWFV会导致细胞收缩、颗粒度增加、凋亡和细胞迁移受损,这些效应可能导致LWFV诱导的组织损伤。

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