宫颈癌细胞分泌外泌体介导上皮-间质转化提高癌前细胞侵袭能力的体外研究

发布时间：2018-05-02 00:40

  本文选题：外泌体 + 宫颈癌　； 参考：《现代妇产科进展》2017年03期

【摘要】：目的:研究宫颈癌细胞Siha分泌的外泌体(Exosomes)能否介导上皮-间质转化(EMT),从而改变癌前细胞Ect1的体外侵袭能力。方法:差速超速离心法分离SihaExo,透射电镜观察形态,Western blot检测标志蛋白表达,共聚焦显微镜观察PKH67染色外泌体的生物学活性。Siha-Exo和PBS分别与Ect1共培养48h后Western blot检测上皮和间质转化标志蛋白的表达,Transwell侵袭实验检测共培养后Ect1体外侵袭能力。结果:Siha-Exo为直径30~100nm的盘状囊泡,特异性表达标志性蛋白CD63和CD81,能够被受体细胞Ect1内吞,具备生物学活性。Western blot结果显示,与PBS组相比,Siha-Exo组下调Ect1细胞中上皮标志蛋白E-Cadherin和β-Catenin的表达,上调间质标志蛋白NCadherin和Vimentin的表达。Transwell侵袭实验显示,与PBS组相比,Siha-Exo组穿透聚碳酯膜的Ect1细胞数明显增多(P=0.01)。结论:Siha-Exo能够介导Ect1发生EMT变化,提高其体外侵袭能力。
[Abstract]:Aim: to investigate whether exocrine exosomes secreted by cervical cancer cells (Siha) can mediate epithelial-interstitial transformation (EMT) and thus change the invasiveness of precancerous Ect1 in vitro. Methods: Siha Exo was isolated by differential ultracentrifugation, and the expression of marker protein was detected by Western blot. Confocal microscopy was used to observe the biological activity of exosomes by PKH67 staining. Siha-Exo and PBS were co-cultured with Ect1 for 48 h. Western blot was used to detect the expression of epithelial and interstitial transforming marker protein. Transwell invasion assay was used to detect the invasiveness of Ect1 after co-culture. Results: Siha-Exo was a disk-shaped vesicle with diameters of 30~100nm, which specifically expressed CD63 and CD81, and could be ingested by receptor cell Ect1. Western blot showed that the expression of E-Cadherin and 尾 -Catenin in Ect1 cells was down-regulated in Siha-Exo group compared with PBS group. Transwell invasion assay showed that compared with PBS group, the number of Ect1 cells in Siha-Exo group penetrating through polycarboxylate membrane was significantly higher than that in PBS group, and the number of Ect1 cells in Siha-Exo group was significantly higher than that in PBS group. Conclusion: Siha-Exo can mediate the change of EMT in Ect1 and improve its invasiveness in vitro.
【作者单位】： 南方医科大学南方医院妇产科;南方医科大学南方医院病理科;
【基金】：国家自然科学基金(No:81072132;81372871;81672589;JCYJ20160429161218745)
【分类号】：R737.33

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