miR-224靶向PIK3CD影响DLBCL细胞增殖和侵袭的机制研究

Mechanism of MiR-224 Affecting DLBCL Cell Proliferation and Invasion by Targeted Inhibition of PIK3CD

目的:探讨miR-224对弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)细胞增殖和侵袭影响的潜在机制。方法:收集2011年6月至2017年12月在昆明医科大学第一附属医院血液科确诊的76例DLBCL初诊患者血液标本(DLBCL组)、41例健康体检者血液标本(正常对照组),以及人淋巴内皮细胞HLEC和DLBCL细胞系HBL1、OCI-LY10、OCI-LY8、OCI-LY19。使用RT-qPCR法检测miR-224和PIK3CD mRNA的表达水平。CCK-8法和克隆形成实验检测HBL1细胞增殖能力,Transwell实验检测HBL1细胞侵袭能力。双荧光素酶报告基因验证miR-224与PIK3CD的靶向关系。Western blot检测PIK3CD蛋白的表达水平。结果:miR-224在DLBCL患者血液中表达显著低于正常对照组(P<0.01);过表达miR-224,HBL1细胞增殖和侵袭能力均显著降低(P<0.01)。PIK3CD是miR-224的靶基因,敲减PIK3CD能显著抑制HBL1细胞的增殖和侵袭能力(P<0.01)。结论:miR-224在DLBCL进展中起着关键的作用,miR-224靶向抑制PIK3CD可降低DLBCL细胞系HBL1细胞的增殖和侵袭能力。

Objective:To investigate the potential mechanisms of miR-224 affecting the proliferation and invasion of diffuse large B-cell lymphoma(DLBCL)cells.Methods:The blood samples of 76 DLBCL patients(DLBCL group)diagnosed at the First Affiliated Hospital of Kunming Medical University from June 2011 to December 2017,and 41 healthy persons of physical examination(normal control group)as well as human lymphatic endothelial cells(HELC)and DLBCL cell lines HBL1,OCI-LY10,OCI-LY8,OCI-LY19 were collected.The expression of miR-224 and PIK3 CD mRNA was measured by RT-qPCR.The proliferation of HBL1 cells was detected by CCK-8 method and colony formation test.The invasion ability of HBL1 cells was detected by Transwell test.Dual-luciferase reporter gene assay was used to verify the relationship between miR-224 and PIK3 CD.The expression of PIK3 CD protein was measured by Western blot.Results:The expression of miR-224 in blood of DLBCL patients was very significantly lower than that in normal control group(P<0.01).The overexpression of miR-224 significantly decreased proliferation and invasion of HBL1 cells(all P<0.01).PIK3 CD was a target gene of miR-224.Knockdown of PIK3 CD significantly suppressed the proliferation and invasion of HBL1 cells(all P<0.01).Conclusion:MiR-224 plays a key role in the progression of DLBCL,and the proliferation and invasion of HBL1 cells can be suppressed by targeted inhibition of PIK3 CD.