中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
7期
1357-1359
,共3页
陈锐涵%朱宝益%叶志强%蔡松旺%蔡燚%叶春伟%陶奕然%温星桥
陳銳涵%硃寶益%葉誌彊%蔡鬆旺%蔡燚%葉春偉%陶奕然%溫星橋
진예함%주보익%협지강%채송왕%채일%협춘위%도혁연%온성교
高糖%前列腺%微小RNA%MiR-301a%增殖
高糖%前列腺%微小RNA%MiR-301a%增殖
고당%전렬선%미소RNA%MiR-301a%증식
High glucose%Prostate%MicroRNA%MiR-301 a%Proliferation
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(＞300 mg/dl),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(50 mmol/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(25～50 mmol/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-186 2.405倍、miR-30la 2.202倍、miR-3652.093倍、miR-193 2.317倍),2个miRNA下调(miR-434 0.298倍、miR-361 0.386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25 mmol/L组、50 mmoL/L组、miR-30la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)；50 mmol/L高糖联合miR-301a抑制物转染组的A值为起始的143％,与50 mmol/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.
目的 檢測高糖刺激下微小RNA(miRNA)在前列腺組織及細胞中的差異錶達,探討高血糖對前列腺影響的分子機製.方法 將成年雄性SD大鼠隨機分為對照組和高血糖組(＞300 mg/dl),應用miRNA芯片技術檢測兩組間miRNA錶達譜的差異,併通過實時定量聚閤酶鏈反應(qRT-PCR)方法驗證；以qRT-PCR檢測差異錶達的miRNAs在高糖(50 mmol/L)培養的RWPE-1細胞中的錶達；噻唑藍(MTT)比色法檢測轉染miRNAs和/或高糖(25～50 mmol/L)培養的RWPE-1的增殖.結果 高血糖組大鼠前列腺組織中4箇miRNA 上調(miR-186 2.405倍、miR-30la 2.202倍、miR-3652.093倍、miR-193 2.317倍),2箇miRNA下調(miR-434 0.298倍、miR-361 0.386倍),差異均有統計學意義(P＜0.05)；高糖培養下,RWPE-1細胞中各miRNAs的錶達趨勢與之一緻.MTT實驗中25 mmol/L組、50 mmoL/L組、miR-30la轉染組的細胞吸光度(A)值分彆為起始的175％、197％、188％,與對照組(122％)比較差異有統計學意義(p＜0.05)；50 mmol/L高糖聯閤miR-301a抑製物轉染組的A值為起始的143％,與50 mmol/L組比較差異有統計學意義(P＜0.05).結論 體內外高糖條件均可引起前列腺miRNA錶達譜變化,高糖通過miR-301a對前列腺上皮細胞的增殖有明顯促進作用.
목적 검측고당자격하미소RNA(miRNA)재전렬선조직급세포중적차이표체,탐토고혈당대전렬선영향적분자궤제.방법 장성년웅성SD대서수궤분위대조조화고혈당조(＞300 mg/dl),응용miRNA심편기술검측량조간miRNA표체보적차이,병통과실시정량취합매련반응(qRT-PCR)방법험증；이qRT-PCR검측차이표체적miRNAs재고당(50 mmol/L)배양적RWPE-1세포중적표체；새서람(MTT)비색법검측전염miRNAs화/혹고당(25～50 mmol/L)배양적RWPE-1적증식.결과 고혈당조대서전렬선조직중4개miRNA 상조(miR-186 2.405배、miR-30la 2.202배、miR-3652.093배、miR-193 2.317배),2개miRNA하조(miR-434 0.298배、miR-361 0.386배),차이균유통계학의의(P＜0.05)；고당배양하,RWPE-1세포중각miRNAs적표체추세여지일치.MTT실험중25 mmol/L조、50 mmoL/L조、miR-30la전염조적세포흡광도(A)치분별위기시적175％、197％、188％,여대조조(122％)비교차이유통계학의의(p＜0.05)；50 mmol/L고당연합miR-301a억제물전염조적A치위기시적143％,여50 mmol/L조비교차이유통계학의의(P＜0.05).결론 체내외고당조건균가인기전렬선miRNA표체보변화,고당통과miR-301a대전렬선상피세포적증식유명현촉진작용.
Objective To study the aberrant expression of miRNAs implicated in the prostate under 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 alteration. Furthermore,these changes were also investigated in the human prostate epithelial cell line RWPE-1 in vitro.Methyl thiazol tetrazolium (MTT) assay was performed to evaluate the influence of high glucose (25-50 mmol/L) and/or miRNAs transfection on the proliferation of RWPE-1. Results Compared 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 miRNAs ( 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 ceils cultured in high-glucose condition.MTT assay showed the cellular absorbance values (A values) of high-glucose groups and miR-30la-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-301 a have positive effect on prostatic epithelium proliferation.
