中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
10期
2070-2072
,共3页
李颖%卢忠心%孔晓宇%魏礼清
李穎%盧忠心%孔曉宇%魏禮清
리영%로충심%공효우%위례청
癌,肝细胞%微小RNA-181b%肝癌缺失基因-1
癌,肝細胞%微小RNA-181b%肝癌缺失基因-1
암,간세포%미소RNA-181b%간암결실기인-1
Carcinoma,hepatocellular%MicroRNA-181b%Deleted in liver cancer-1
目的 探讨微小RNA(miRNA,miR)-181b对肝癌缺失基因-1(DLC-1)的调控及其在肝癌发病中的作用.方法 通过实时荧光定量聚合酶链反应(FQ-PCR)和Western blot检测4份正常肝脏组织,17份癌旁组织、肝癌组织及肝癌SMMC7721细胞、胚肝LO2细胞中miR-181b和DLC-1的mRNA和蛋白质表达水平.应用miR-181b抑制剂下调肝癌细胞SMMC7721中miR-181b表达后,实时定量聚合酶链反应和Westem blot检测SMMC7721细胞中DLC-1表达水平的变化.结果 miR-181b在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.162 ±0.004、0.175±0.021和0.656±0.034,肝癌组织高于正常肝组织(P＜0.05).miR-181b在SMMC7721细胞中的表达较LO2细胞上调(0.723±0.042和0.262±0.037,P＜0.01).DLC-1 mRNA在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.392±0.094、0.187 ±0.043和0.081 ±0.012,肝癌组织低于正常肝组织(P＜0.05)；DLC-1蛋白在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.423±0.026、0.214±0.029和0.112±0.021,差异有统计学意义(P＜0.01).miR-181b抑制剂转染SMMC7721细胞后,DLC-1蛋白在空白组、转染组和阴性对照组的相对表达量分别为0.132±0.027、0.379±0.019和0.125±0.015,转染组较其他两组明显升高(P＜0.05).结论 miR-181b在肝癌组织中表达上调,其抑制DLC-1的表达可能是肝癌发生的重要机制.
目的 探討微小RNA(miRNA,miR)-181b對肝癌缺失基因-1(DLC-1)的調控及其在肝癌髮病中的作用.方法 通過實時熒光定量聚閤酶鏈反應(FQ-PCR)和Western blot檢測4份正常肝髒組織,17份癌徬組織、肝癌組織及肝癌SMMC7721細胞、胚肝LO2細胞中miR-181b和DLC-1的mRNA和蛋白質錶達水平.應用miR-181b抑製劑下調肝癌細胞SMMC7721中miR-181b錶達後,實時定量聚閤酶鏈反應和Westem blot檢測SMMC7721細胞中DLC-1錶達水平的變化.結果 miR-181b在正常肝組織、癌徬組織和肝癌組織中的相對錶達量分彆為0.162 ±0.004、0.175±0.021和0.656±0.034,肝癌組織高于正常肝組織(P＜0.05).miR-181b在SMMC7721細胞中的錶達較LO2細胞上調(0.723±0.042和0.262±0.037,P＜0.01).DLC-1 mRNA在正常肝組織、癌徬組織和肝癌組織中的相對錶達量分彆為0.392±0.094、0.187 ±0.043和0.081 ±0.012,肝癌組織低于正常肝組織(P＜0.05)；DLC-1蛋白在正常肝組織、癌徬組織和肝癌組織中的相對錶達量分彆為0.423±0.026、0.214±0.029和0.112±0.021,差異有統計學意義(P＜0.01).miR-181b抑製劑轉染SMMC7721細胞後,DLC-1蛋白在空白組、轉染組和陰性對照組的相對錶達量分彆為0.132±0.027、0.379±0.019和0.125±0.015,轉染組較其他兩組明顯升高(P＜0.05).結論 miR-181b在肝癌組織中錶達上調,其抑製DLC-1的錶達可能是肝癌髮生的重要機製.
목적 탐토미소RNA(miRNA,miR)-181b대간암결실기인-1(DLC-1)적조공급기재간암발병중적작용.방법 통과실시형광정량취합매련반응(FQ-PCR)화Western blot검측4빈정상간장조직,17빈암방조직、간암조직급간암SMMC7721세포、배간LO2세포중miR-181b화DLC-1적mRNA화단백질표체수평.응용miR-181b억제제하조간암세포SMMC7721중miR-181b표체후,실시정량취합매련반응화Westem blot검측SMMC7721세포중DLC-1표체수평적변화.결과 miR-181b재정상간조직、암방조직화간암조직중적상대표체량분별위0.162 ±0.004、0.175±0.021화0.656±0.034,간암조직고우정상간조직(P＜0.05).miR-181b재SMMC7721세포중적표체교LO2세포상조(0.723±0.042화0.262±0.037,P＜0.01).DLC-1 mRNA재정상간조직、암방조직화간암조직중적상대표체량분별위0.392±0.094、0.187 ±0.043화0.081 ±0.012,간암조직저우정상간조직(P＜0.05)；DLC-1단백재정상간조직、암방조직화간암조직중적상대표체량분별위0.423±0.026、0.214±0.029화0.112±0.021,차이유통계학의의(P＜0.01).miR-181b억제제전염SMMC7721세포후,DLC-1단백재공백조、전염조화음성대조조적상대표체량분별위0.132±0.027、0.379±0.019화0.125±0.015,전염조교기타량조명현승고(P＜0.05).결론 miR-181b재간암조직중표체상조,기억제DLC-1적표체가능시간암발생적중요궤제.
Objective To investigate the regulatory effect ofmicroRNA-181b (miR-181b) on deleted in liver cancer-1 (DLC-1) and its role in hepatocarcinogenesis.Methods miR-181b and DLC-1 mRNA expression levels in normal tissue,paraneoplastic tissue,liver cancer tissue and liver cancer cells were detected by using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).DLC-1 protein expression was detected by using Western blotting.miR-181b inhibitor was transfected into SMMC7721 cells and the expression changes of DLC-1 mRNA and protein were examined by using FQ-PCR and Western blotting respectively.Results miR-181b was up-regulated in hepatocellular carcinoma and adjacent tissues as compared to normal liver tissues and the expression of miR-181 b was also increased in SMMC7721 cells as compared to fetal liver cells LO2,whereas the expression levels of DLC-1 mRNA and protein were decreased in paraneoplastic and HCC tissues compared with normal liver tissues.The expression of miR-181b and DLC-1 was negatively correlated.No obvious difference was found in the DLC-1 mRNA expression after miR-181b inhibitor transfection.Conclusion The DLC-1 abnormal low-expression may play a dynamic role in hepatocarcinogenesis due to the miR-181 b up-regulation.
