中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
9期
1927-1929
,共3页
蔡燚%李小娟%朱宝益%陶奕然%李骏%蔡松旺%陈锐涵%温星桥
蔡燚%李小娟%硃寶益%陶奕然%李駿%蔡鬆旺%陳銳涵%溫星橋
채일%리소연%주보익%도혁연%리준%채송왕%진예함%온성교
高糖%微小RNA-301a%前列腺癌%细胞周期
高糖%微小RNA-301a%前列腺癌%細胞週期
고당%미소RNA-301a%전렬선암%세포주기
High glucose%microRNA-301a%Prostate cancer%Cell cycle
目的 探讨微小RNA(miR)-301a介导高糖促进前列腺癌细胞生长的分子机制.方法 通过实时定量聚合酶链反应(Real-time PCR)检测miR-301a的表达.细胞计数试剂盒(CCK-8)法检测转染miRNAs和/或高糖(4.5 g/L)培养的前列腺癌DU145细胞的增殖.应用微管聚合抑制剂诺考达唑(Nocodazole,40 μg/L)处理16h或者血清饥饿处理48 h使细胞周期同步化,流式细胞仪分析miR-301a对细胞周期时相转换的影响.CCK-8法测定细胞生长活性,检测高糖及miR-301a对前列腺癌细胞增殖的影响.结果 miR-301a在前列腺癌细胞PC3、DU145的表达分别为正常前列腺上皮细胞RWPE-1的(2.86±0.75)倍和(4.05±0.96)倍.高糖培养DU145细胞24、48 h后miR-301a的表达分别为对照组的(1.88±0.34)倍和(13.15 ±3.45)倍.抑制miR-301a可部分逆转高糖的促增殖作用.过表达miR-301a后经细胞周期同步化处理,与对照组比较miR-301a可促进前列腺癌DU145细胞G1/S时相转换.结论 miR-301a在前列腺癌细胞中表达上调,miR-301a介导高糖促进前列腺细胞周期G1/S时相转换,促进细胞增殖.
目的 探討微小RNA(miR)-301a介導高糖促進前列腺癌細胞生長的分子機製.方法 通過實時定量聚閤酶鏈反應(Real-time PCR)檢測miR-301a的錶達.細胞計數試劑盒(CCK-8)法檢測轉染miRNAs和/或高糖(4.5 g/L)培養的前列腺癌DU145細胞的增殖.應用微管聚閤抑製劑諾攷達唑(Nocodazole,40 μg/L)處理16h或者血清饑餓處理48 h使細胞週期同步化,流式細胞儀分析miR-301a對細胞週期時相轉換的影響.CCK-8法測定細胞生長活性,檢測高糖及miR-301a對前列腺癌細胞增殖的影響.結果 miR-301a在前列腺癌細胞PC3、DU145的錶達分彆為正常前列腺上皮細胞RWPE-1的(2.86±0.75)倍和(4.05±0.96)倍.高糖培養DU145細胞24、48 h後miR-301a的錶達分彆為對照組的(1.88±0.34)倍和(13.15 ±3.45)倍.抑製miR-301a可部分逆轉高糖的促增殖作用.過錶達miR-301a後經細胞週期同步化處理,與對照組比較miR-301a可促進前列腺癌DU145細胞G1/S時相轉換.結論 miR-301a在前列腺癌細胞中錶達上調,miR-301a介導高糖促進前列腺細胞週期G1/S時相轉換,促進細胞增殖.
목적 탐토미소RNA(miR)-301a개도고당촉진전렬선암세포생장적분자궤제.방법 통과실시정량취합매련반응(Real-time PCR)검측miR-301a적표체.세포계수시제합(CCK-8)법검측전염miRNAs화/혹고당(4.5 g/L)배양적전렬선암DU145세포적증식.응용미관취합억제제낙고체서(Nocodazole,40 μg/L)처리16h혹자혈청기아처리48 h사세포주기동보화,류식세포의분석miR-301a대세포주기시상전환적영향.CCK-8법측정세포생장활성,검측고당급miR-301a대전렬선암세포증식적영향.결과 miR-301a재전렬선암세포PC3、DU145적표체분별위정상전렬선상피세포RWPE-1적(2.86±0.75)배화(4.05±0.96)배.고당배양DU145세포24、48 h후miR-301a적표체분별위대조조적(1.88±0.34)배화(13.15 ±3.45)배.억제miR-301a가부분역전고당적촉증식작용.과표체miR-301a후경세포주기동보화처리,여대조조비교miR-301a가촉진전렬선암DU145세포G1/S시상전환.결론 miR-301a재전렬선암세포중표체상조,miR-301a개도고당촉진전렬선세포주기G1/S시상전환,촉진세포증식.
Objective To investigate the molecular mechanism of microRNA (miR)-301a in the regulation prostate cancer cell proliferation by high glucose.Methods The expression of miR-301a was detected by real-time quantitative polymerase chain reaction (Real-time PCR) (Real-time PCR).cell counting kit-8 (CCK-8) assay was performed to evaluate the influence of high glucose (4.5 g/L) and/or miRNAs transfection on the proliferation of DU145.Nocodazole-treated for 16 hours or serum-starved for 48 hours were used to synchronized cell cycle,the transition of cell cycle was detected by flow cytometry.Results The expression level of miR-301a in PC3 and DU145 cells were 2.86 ±0.75 and 4.05 ±0.96 times higher than the RWPE-1 cells respectively.The expression of miR-301a were 1.88 ± 0.34 and 13.15 ±3.45 times higher than control group in DU145 cells cultured in high glucose for 24 and 48 hour.Inhibition of miR-301 a could reverse the effect on proliferation of high glucose.Overexpression of miR-301 a promoted G1/S phase transition in prostate cancer DU145 cells.Conclusion miR-301a was up-reglulated in prostate cancer cell lines,which mediates G1/S cell cycle phase transition promoted by high glucose.
