miR-20a-5p表达对胃粘膜相关组织淋巴瘤增殖的影响及其机制研究  

The study on mechanism of miR-20a-5p on cell proliferation in gastric MALT lymphoma

作　　者：农卫霞[1] 王奎 NONG Weixia;WANG Kui(The First Affiliated Hospital,Shool of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China;School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学医学院第一附属医院,新疆石河子832000 [2]石河子大学医学院,新疆石河子832000

出　　处：《石河子大学学报（自然科学版）》2021年第1期102-108,共7页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(81860374)。

摘　　要：目的探讨miR-20a-5p的表达对胃粘膜相关组织(MALT)淋巴瘤的增殖影响及其相关分子机制。方法收集18例人胃MALT淋巴瘤组织及邻近正常胃粘膜组织。采用实时荧光定量聚合酶链反应(qRT-PCR)检测组织中miR-20a-5p的相对表达;应用miR-20a-5p mimic和miR-20a-5p inhibitor分别转染Raji淋巴瘤细胞,以细胞增殖活性(CCK-8)实验分析miR-20a-5p对细胞的增殖的影响;通过计算机靶基因预测软件Target Scan、双荧光素酶报告基因实验及Western blot实验确定miR-20a-5p作用的可能靶基因;实时荧光定量PCR(qRT-PCR)检测组织中runt相关转录因子3(RUNX3)的相对表达量。结果与邻近正常粘膜组织相比,胃MALT淋巴瘤组织中miR-20a-5p的表达异常增高(P<0.05)。CCK-8实验显示,与对照组相比,miR-20a-5p mimic转染Raji淋巴瘤细胞的吸光度显著升高(P<0.05),miR-20a-5p inhibitor组吸光度显著降低(P<0.05)。生物信息学软件预测RUNX3为miR-20a-5p作用的靶基因。双荧光素酶报告实验证明miR-20a-5p与RUNX3的3’UTR区特异性结合。Western Blot实验结果显示miR-20a-5p mimic组RUNX3蛋白表达水平明显低于对照组(P<0.05),miR-20a-5p inhibitor组RUNX3蛋白表达水平显著高于对照组(P<0.05)。MALT淋巴瘤患者标本中的RUNX3 mRNA水平均显著低于邻近胃粘膜组织(P<0.05)。人MALT淋巴瘤组织中miR-20a-5p表达与RUNX3 mRNA表达呈负相关关系(P<0.05)。结论 miR-20a-5p可促进Raji淋巴瘤细胞的增殖,其机制与下调RUNX3的表达有关。miR-20-5p在胃MALT呈低表达,与RUNX3表达呈负相关关系,miR-20-5p为MALT淋巴瘤的临床治疗提供新靶标。Objective To investigate the role of miR-20 a-5 p on proliferation of gastric mucosal associated lymphoid tissue( MALT)lymphoma and explored its related molecular mechanism. Method 18 cases patients MALT lymphoma and adjacent normal mucosal tissue were collected. The relative expression of miR-20 a-5 p in gastric MALT lymphoma tissues and adjacent tissues was detected by quantitative real-time polymerase chain reaction( qRT-PCR). Raji cells were transfected with miR-20 a-5 p mimic and inhibitor for investigating the proliferation by cell counting kit-8( CCK-8). Computer software target Scan,luciferase assays and western blot were performed to verify the direct target of miR-20 a-5 p,and the target genes were preliminarily verified. Runt related transcription factor 3 gene( RUNX3) mRNA expression level in gastric MALT lymphoma tissues and adjacent tissues was detected by quantitative Real-Time PCR( qRT-PCR). Results Compared with adjacent normal mucosa tissues,the expression level of miR-20 a-5 p in gastric MALT lymphoma tissues was significantly increased( P<0. 05). Compared with the adjacent normal mucosal tissues,the expression level of miR-20 a-5 p in gastric MALT lymphoma tissues was abnormally increased( P < 0. 05). CCK-8 results showed that the absorbance of Raji cells with miR-20 a-5 p mimic was significantly increased compared with the control group( P<0. 05),the absorbance of miR-20 a-5 p inhibitor group was significantly decreased( P<0. 05). RUNX3 was identified as the potential target of miR-20 a-5 p by bioinformatics software. Dual-luciferase assays showed that miR-20 a-5 p could bond with 3’UTR regions of RUNX3. Western Blotting showed that RUNX3 protein expression in the miR-20 a-5 p inhibitor group was significantly lower than that in the negative control( P<0. 05). The mRNA levels of RUNX3 in the samples of patients with gastric MALT lymphoma were significantly lower than those in the adjacent gastric mucosal tissues( P<0. 05). Correction analysis showed that miR-20 a-5 p expression was inversel

关 键 词：粘膜相关组织淋巴瘤 miR-20a-5p RAJI细胞 增殖 RUNT相关转录因子3 

分 类 号：R733.1[医药卫生—肿瘤]