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。
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.