目的 探讨血浆外泌体中微RNA(micro RNA,miR)-21对弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)细胞生长和转移的影响及机制.方法 提取并鉴定DLBCL患者血浆外泌体,将U2932细胞分为miR-21模拟阴性对照(mimic negative control,NC)组、miR-21组、miR-21+人第10号染色体缺失的磷酸酶及张力蛋白同源(phosphatase and tensin homolog deleted on chromosome ten,PTEN)组、磷酸盐缓冲液(phosphate buffer solution,PBS)组、DLBCL 患者外泌体组、DLBCL患者外泌体+miR-21抑制剂组(A组)以及DLBCL患者外泌体+PTEN组(B组).采用实时荧光定量聚合酶链反应检测U2932细胞中miR-21表达;采用双荧光素酶验证miR-21与PTEN的靶向关系;采用细胞计数试剂盒-8和Transwell检测各组U2932细胞增殖、迁移和侵袭能力.结果 DLBCL患者外泌体为双层膜,且形态呈"茶托"样,粒径30～100 nm,表面表达标志蛋白肿瘤易感基因101、分化抗原9以及热激蛋白70.双荧光素酶结果显示PTEN为miR-21的靶基因.与miR-NC组相比,miR-21组U2932细胞的增殖、迁移和侵袭能力显著增强(P＜0.05);与miR-21组比较,miR-21+PTEN组U2932细胞增殖、迁移和侵袭能力均降低(P＜0.05).与PBS组比较,DLBCL患者血浆外泌体组U2932细胞中miR-21表达增加,细胞增殖、迁移和侵袭能力增强(P＜0.05);A、B组U2932细胞增殖、迁移和侵袭能力低于DLBCL患者血浆外泌体组(P＜0.05).结论 血浆外泌体miR-21可抑制PTEN,促进U2932细胞的增殖、迁移和侵袭.
Effects of plasma exosomal micro RNA-21 on the growth and metastasis of diffuse large B-cell lymphoma cells
Objective To investigate the effects of plasma exosomal micro RNA(miR)-21 on the growth and metastasis of diffuse large B-cell lymphoma(DLBCL)cells and its mechanism.Methods Extraction and identification of plasma exosomes from DLBCL patients.U2932 cells were divided into miR-21 mimic negative control(NC)group,miR-21 group,miR-21+phosphatase and tensin homolog deleted on chromosome ten(PTEN)group,phosphate buffer solution(PBS)group,DLBCL exosome group,DLBCL exosome+miR-21 inhibitor group(A group),DLBCL exosome+PTEN group(B group).Real time fluorescent quantitative polymerase chain reaction was used to detect miR-21 expression in U2932 cells.Dual luciferase was used to verify the targeting relationship between miR-21 and PTEN.The proliferation,migration and invasion ability of U2932 cells were detected by cell counting kit-8 and Transwell assay.Results The exosomes of DLBCL patients had a bilayer membranes with a"saucer-like"shape,with a particle size of 30～100 nm.The marker proteins tumor susceptibility gene 101,cluster of differentiation 9 and heat shock protein 70 were expressed on the surface.Dual luciferase reporter gene assay showed that PTEN was the target gene of miR-21.Compared with the miR-NC group,the proliferation,migration and invasion abilities of the U2932 cells in the miR-21 group were significantly increased(P＜0.05).Compared with miR-21 group,the proliferation,migration and invasion ability of U2932 cells in miR-21+PTEN group were significantly decreased(P＜0.05).Compared with PBS group,the expression of miR-21 in plasma exosome U2932 cells of DLBCL patients was significantly increased,and cell proliferation,migration and invasion were significantly enhanced(P＜0.05).The proliferation,migration and invasion ability of U2932 cells in A and B groups were significantly lower than those in the plasma exosome group of DLBCL patients(P＜0.05).Conclusion The plasma exosome miR-21 promotes the proliferation,migration and invasion of U2932 cells by inhibiting PTEN.
plasma exosomemicro RNA-21diffuse large B-cell lymphomagrowth and metastasis
NETL
NSTL
万方数据
