[1]刘小豪,王方圆,侯晋,等.SACC-83来源的外泌体促进腺样囊性癌细胞的增殖[J].南方医科大学学报,2018,(08):1008.
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SACC-83来源的外泌体促进腺样囊性癌细胞的增殖()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年08期
页码:
1008
栏目:
出版日期:
2018-07-31

文章信息/Info

Title:
Adenoid cystic carcinoma cells produce exosomes that promote tumor cell proliferation
作者:
刘小豪王方圆侯晋张磊涛陈之锋殷学民
关键词:
腺样囊性癌外泌体细胞增殖ERK/P-ERK
Keywords:
adenoid cystic carcinoma exosomes cell proliferation ERK/P-ERK
摘要:
目的研究腺样囊性癌(ACC)肿瘤细胞来源的外泌体对自身肿瘤细胞增殖能力的影响。方法选取ACC细胞株SACC- 83,采用超滤管浓缩与试剂盒提取相结合的方法从SACC-83的培养上清中提取外泌体,通过透射电镜及蛋白免疫印迹法对所 提取的外泌体进行鉴定;将SACC-83来源的外泌体用荧光染料PKH67标记后与其分泌细胞共培养,采用激光扫描共聚焦显微 镜(LSCM)观察SACC-83对于自身来源的外泌体的摄取情况;采用MTT细胞增殖实验、细胞划痕实验及Western blot 法检测 SACC-83经自身外泌体作用后,其细胞增殖能力的变化情况以及ERK/P-ERK蛋白的表达变化。结果透射电镜及Western blot 的检测结果显示,SACC-83培养上清中所提取到的微囊泡直径为30~100 nm,且外泌体蛋白标记物CD9、CD63和TSG101的表 达为阳性;携带PKH67荧光标记的外泌体可被SACC-83摄取。MTT及细胞划痕实验结果示SACC-83来源的外泌体可促进自 身肿瘤细胞的增殖能力,Western blot结果显示P-ERK的表达上调,ImageJ软件行蛋白条带灰度值分析并统计显示实验组与对 照组具有显著统计学差异(P<0.05)。结论SACC-83来源的外泌体可被自身肿瘤细胞摄取,并可显著促进肿瘤细胞的增殖能 力,上调P-ERK的表达。该结果提示ACC可能通过外泌体途径促进肿瘤细胞的增殖能力,且ERK的活化以及MAPK/ERK信 号通路的激活可能是其促细胞增殖的潜在机制之一。
Abstract:
Objective To observe the effect of exosomes released by adenoid cystic carcinoma (ACC) cell line SACC-83 on the proliferation of ACC cells. Methods Exosomes were isolated from SACC-83 cell culture supernatants using total exosome isolation reagents. The whole-mount exosomes were characterized using transmission electron microscope and Western blotting. The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with SACC-83 cells for 48 h, followed by staining with Alexa Fluor 594 phalloidin and DAPI to observe exosome uptake by the cells using laser scanning confocal microscopy (LSCM). The cell proliferation was assessed using MTT assay and wound healing assay, and the expressions of ERK and P-ERK in the co-cultured SACC-83 cells were detected using Western blotting. Results The exosomes isolated from SACC-83 cells showed a size range of 30-100 nm and expressed the exosomal markers CD9, CD63 and TSG101. LSCM showed exosome uptake by SACC-83 cells, which exhibited accelerated proliferation and significantly enhanced P-ERK expression (P< 0.05) without significant changes in ERK expression. Conclusion SACC-83 cells produce exosomes that promote the tumor cell proliferation and enhances the cellular expression of P-ERK, suggesting a potential role of MAPK/ERK pathway activation in exosome-mediated acceleration of ACC cell proliferation.

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更新日期/Last Update: 1900-01-01