中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
10期
2108-2111
,共4页
肝细胞肝癌%肝癌血管内皮细胞%微小RNA-1%RNA干扰
肝細胞肝癌%肝癌血管內皮細胞%微小RNA-1%RNA榦擾
간세포간암%간암혈관내피세포%미소RNA-1%RNA간우
Hepatocellular carcinoma%Vascular endothelial cells of human hepatocellular carcinoma%MicroRNA-1%RNA interference
目的 通过构建慢病毒介导的微小RNA(miRNA)-1-小干扰RAN (siRNA),观察下调miR-1表达对人肝癌血管内皮细胞(TEC)生物学行为的影响.方法 针对目标序列合成特异性siRNA干扰片段并克隆入慢病毒载体,将重组miRNA-1-siRNA-GV159转染TEC,实验分为3组:(1)下调组(miRNA-1-siRNA,MD组);(2)阴性对照组(NC组);(3)空白对照组(CON组).分别用噻唑蓝(MTT)法检测细胞增殖,流式细胞仪检测细胞的凋亡及细胞周期,划痕实验检测细胞的迁移能力.结果 MTT结果显示MD组细胞在转染miRNA-1-siRNA-GV159后的第1、2、3、4、5天5个时间点均表现出吸光度值降低,与NC组及CON组比较差异有统计学意义(P＜0.05);流式细胞仪检测凋亡结果显示CON、NC、MD组凋亡率分别为(2.03±0.30)％、(2.18±0.15)％、(6.32±0.33)％,MD组细胞凋亡率比其他两组明显明显增加,差异有统计学意义(P＜0.05);而细胞周期检测结果显示3组之间差异无统计学意义(P＞0.05);划痕实验结果显示划痕12h后CON、NC、MD3组细胞的迁移距离分别为(270.6 ±44.5)、(299.5 ±28.7)、(198.4 ±49.6)μm,MD组和其他两组比较迁移能力明显下降,差异有统计学意义(P＜0.05).结论 下调miRNA-1表达可抑制TEC的增殖能力,增加TEC的凋亡并降低其迁移能力.
目的 通過構建慢病毒介導的微小RNA(miRNA)-1-小榦擾RAN (siRNA),觀察下調miR-1錶達對人肝癌血管內皮細胞(TEC)生物學行為的影響.方法 針對目標序列閤成特異性siRNA榦擾片段併剋隆入慢病毒載體,將重組miRNA-1-siRNA-GV159轉染TEC,實驗分為3組:(1)下調組(miRNA-1-siRNA,MD組);(2)陰性對照組(NC組);(3)空白對照組(CON組).分彆用噻唑藍(MTT)法檢測細胞增殖,流式細胞儀檢測細胞的凋亡及細胞週期,劃痕實驗檢測細胞的遷移能力.結果 MTT結果顯示MD組細胞在轉染miRNA-1-siRNA-GV159後的第1、2、3、4、5天5箇時間點均錶現齣吸光度值降低,與NC組及CON組比較差異有統計學意義(P＜0.05);流式細胞儀檢測凋亡結果顯示CON、NC、MD組凋亡率分彆為(2.03±0.30)％、(2.18±0.15)％、(6.32±0.33)％,MD組細胞凋亡率比其他兩組明顯明顯增加,差異有統計學意義(P＜0.05);而細胞週期檢測結果顯示3組之間差異無統計學意義(P＞0.05);劃痕實驗結果顯示劃痕12h後CON、NC、MD3組細胞的遷移距離分彆為(270.6 ±44.5)、(299.5 ±28.7)、(198.4 ±49.6)μm,MD組和其他兩組比較遷移能力明顯下降,差異有統計學意義(P＜0.05).結論 下調miRNA-1錶達可抑製TEC的增殖能力,增加TEC的凋亡併降低其遷移能力.
목적 통과구건만병독개도적미소RNA(miRNA)-1-소간우RAN (siRNA),관찰하조miR-1표체대인간암혈관내피세포(TEC)생물학행위적영향.방법 침대목표서렬합성특이성siRNA간우편단병극륭입만병독재체,장중조miRNA-1-siRNA-GV159전염TEC,실험분위3조:(1)하조조(miRNA-1-siRNA,MD조);(2)음성대조조(NC조);(3)공백대조조(CON조).분별용새서람(MTT)법검측세포증식,류식세포의검측세포적조망급세포주기,화흔실험검측세포적천이능력.결과 MTT결과현시MD조세포재전염miRNA-1-siRNA-GV159후적제1、2、3、4、5천5개시간점균표현출흡광도치강저,여NC조급CON조비교차이유통계학의의(P＜0.05);류식세포의검측조망결과현시CON、NC、MD조조망솔분별위(2.03±0.30)％、(2.18±0.15)％、(6.32±0.33)％,MD조세포조망솔비기타량조명현명현증가,차이유통계학의의(P＜0.05);이세포주기검측결과현시3조지간차이무통계학의의(P＞0.05);화흔실험결과현시화흔12h후CON、NC、MD3조세포적천이거리분별위(270.6 ±44.5)、(299.5 ±28.7)、(198.4 ±49.6)μm,MD조화기타량조비교천이능력명현하강,차이유통계학의의(P＜0.05).결론 하조miRNA-1표체가억제TEC적증식능력,증가TEC적조망병강저기천이능력.
Objective To study the effects of suppressing the expression of microRNA (miRNA)-1 by lentivirus-mediated miRNA-1-siRNA on biological behaviors in the vascular endothelial cells of human hepatocellular carcinoma (TEC).Methods miRNA-1 specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector.TEC were infected by miRNA-1-siRNA recombinant lentivirus (miRNA-1-siRNA-GV159).TEC cells were divided into miRNA-1 down-regulation (MD) group,negative control (NC) group,control (CON) group.The proliferation of TEC were detected by thiazolyl blue (MTT) assay,the cell cycle and the apoptosis of TEC were detected by flow cytometry,and the cell migration was assessed by a wound healing assay.Results The results of MTT showed that at 1,2,3,4,5 d after infected by miRNA-1-siRNA-GV159 the optical density of TEC in the MD group were all significantly lower than the other two groups (P ＜ 0.05).The results of flow eytometry showed that the apoptosis rate of TEC in CON group,NC group,MD group was (2.03 ±0.30)％,(2.18±0.15)％,(6.32 ±0.33)％ respectively.The apoptosis rate of TEC in MD group was significantly increased compared with the other two group (P ＜ 0.05).However,there was no obvious differences among the there groups on the cell cycle of TEC after suppressing the expression of miRNA-1 (P ＞ 0.05).The results of wound healing assay showed that 12 h after scratching the distance of migration of TEC in CON group,NC group,MD group was (270.6 ± 44.5),(299.5 ± 28.7),(198.4 ± 49.6) μm respectively.The distance of migration in MD group was significantly decreased conpared with the other groups (P ＜ 0.05).Conclusion Suppressing the expression of miRNA-1 can suppress the proliferation,increase the apoptosis and decrease the migration of TEC.
