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高糖诱导前列腺上皮细胞微小RNA的差异表达及miR-301a的促增殖作用
  • ISSN号:1001-9030
  • 期刊名称:《中华实验外科杂志》
  • 分类:R446.62[医药卫生—诊断学;医药卫生—临床医学]
  • 作者机构:[1]中山大学附属第三医院急诊外科,广州510630, [2]中山大学附属第三医院泌尿外科,广州510630, [3]中山大学附属第三医院心胸外科,广州510630
  • 相关基金:国家自然科学基金资助项目(30872585,30901768);985创新团队项目(82000-3281901);2011年教育部新世纪人才支持计划项目
中文摘要:

目的检测高糖刺激下微小RNA(miRNA)在前列腺组织及细胞中的差异表达,探讨高血糖对前列腺影响的分子机制。方法将成年雄性SD大鼠随机分为对照组和高血糖组(〉300mg/d1),应用miRNA芯片技术检测两组间miRNA表达谱的差异,并通过实时定量聚合酶链反应(qRT—PCR)方法验证;以qRT.PCR检测差异表达的miRNAs在高糖(50mmoL/L)培养的RWPE-1细胞中的表达;噻唑蓝(MTT)比色法检测转染miRNAs和/或高糖(25~50mmol/L)培养的RWPE—1的增殖。结果高血糖组大鼠前列腺组织中4个miRNA上调(miR-1862.405倍、miR-301a2.202倍、miR-3652.093倍、miR-1932.317倍),2个miRNA下调(miR-4340.298倍、miR-3610.386倍),差异均有统计学意义(P〈0.05);高糖培养下,RWPE-1细胞中各miRNAs的表达趋势与之一致。MTT实验中25mmol/L组、50mmol/L组、miR-301a转染组的细胞吸光度(A)值分别为起始的175%、197%、188%,与对照组(122%)比较差异有统计学意义(P〈0.05);50mmol/L高糖联合miR-301a抑制物转染组的A值为起始的143%,与50mmol/L组比较差异有统计学意义(P〈0.05)。结论体内外高糖条件均可引起前列腺miRNA表达谱变化,高糖通过miR-301a对前列腺上皮细胞的增殖有明显促进作用。

英文摘要:

Objective To study the aberrant expression of miRNAs implicated in the prostate un- der high-glucose treatment and to uncover the molecular impact of hyperglycemia exerted on the prostate. Methods A hyperglycemia rat model was induced by intraperitoneal injection of streptozotocin on male Sprague-Dawley rats. MicroRNA array was taken to detect the different expression of miRNAs in prostate tissues of rats, with quantitative real time polymerase chain reaction (qRT-PCR) adapted to confirm the al- teration. Furthermore, these changes were also investigated in the human prostate epithelial cell line RWPE-1 in vitro. Methyl thiazol tetrazolium (MTF) assay was performed to evaluate the influence of high glucose (25-50 mmo]/L) and/or miRNAs transfeetion on the proliferation of RWPE-1. Results Com- pared to the control, four miRNAs were significantly up-regulated (miR-186 240.5%, miR-301a 220. 2% , miR-365 209. 3% , miR-193 231.7% )in the hyperglycemia group (P 〈 0.05 ), while two miR- NAs ( miR-434 29. 8% , miR-361 38.6% ) were obviously down-regulated (P 〈 0. 05). These changes were consistent with the qRT-PCR results in RWPE-1 cells cultured in high-glucose condition. MTT assay showed the cellular absorbance values (A values ) of high-glucose groups and miR-301a-transfeeted group increased by 75 % , 97 % and 88% respectively at the 48th hour, while 22% for the control ( P 〈 0. 05). Meanwhile, the A value of the 50 mmol/L group with miR-301 a-inhibitor transfected was significantly less than that of the 50 mmol/L group (P 〈 0. 05 ). Conclusion Differential expression of miRNAs could be induced in prostate by high-glucose condition both in vitro and in vivo. Hyperglycemia and miR-301a have positive effect on prostatic epithelium proliferation.

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期刊信息
  • 《中华实验外科杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中华医学会
  • 主编:
  • 地址:武汉市武昌区东湖路165号
  • 邮编:430064
  • 邮箱:cjes@cma.org.cn
  • 电话:027-87893475
  • 国际标准刊号:ISSN:1001-9030
  • 国内统一刊号:ISSN:42-1213/R
  • 邮发代号:38-85
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:34996