基础医学与临床
基礎醫學與臨床
기출의학여림상
Basic & Clinical Medicine
2015年
2期
213-217
,共5页
口腔鳞状细胞癌%miR-141%EphA2%增殖
口腔鱗狀細胞癌%miR-141%EphA2%增殖
구강린상세포암%miR-141%EphA2%증식
oral squamous cell carcinoma%miR-141%EphA2%proliferation
目的：探讨miR-141对口腔鳞状细胞癌（ OSCC） CAL27细胞增殖活性的影响及其与EphA2的靶向关系。方法构建pre-miR-141、EphA2野生型（EphA2-WT）和突变型（EphA2-MT）真核表达载体，采用实时定量PCR（qRT-PCR）检测pre-miR-141的转染效率。用MTT检测细胞增殖活性，用双荧光素酶报告基因检测、qRT-PCR和Western blot验证miR-141与EphA2的靶向关系。结果 pcDNATM6.2-GW-pre-miR-141、pmirGLO-EphA2-WT和pmirGLO-E-phA2-MT表达载体构建成功，转染pre-miR-141的CAL27细胞miR-141的表达显著升高（ P＜0.001）。 miR-141过表达能够明显抑制CAL27细胞的增殖（ P＜0.05），共转染pre-miR-141和EphA2-WT的CAL 27细胞其荧光活性显著下降（P＜0.001），转染pre-miR-141的CAL27细胞其EphA2的表达要显著低于转染空质粒组（P＜0.001），miR-141的过表达能够显著抑制EphA2的蛋白表达水平。结论 miR-141的过表达可能通过靶向下调EphA2的表达从而抑制OSCC的增殖。
目的：探討miR-141對口腔鱗狀細胞癌（ OSCC） CAL27細胞增殖活性的影響及其與EphA2的靶嚮關繫。方法構建pre-miR-141、EphA2野生型（EphA2-WT）和突變型（EphA2-MT）真覈錶達載體，採用實時定量PCR（qRT-PCR）檢測pre-miR-141的轉染效率。用MTT檢測細胞增殖活性，用雙熒光素酶報告基因檢測、qRT-PCR和Western blot驗證miR-141與EphA2的靶嚮關繫。結果 pcDNATM6.2-GW-pre-miR-141、pmirGLO-EphA2-WT和pmirGLO-E-phA2-MT錶達載體構建成功，轉染pre-miR-141的CAL27細胞miR-141的錶達顯著升高（ P＜0.001）。 miR-141過錶達能夠明顯抑製CAL27細胞的增殖（ P＜0.05），共轉染pre-miR-141和EphA2-WT的CAL 27細胞其熒光活性顯著下降（P＜0.001），轉染pre-miR-141的CAL27細胞其EphA2的錶達要顯著低于轉染空質粒組（P＜0.001），miR-141的過錶達能夠顯著抑製EphA2的蛋白錶達水平。結論 miR-141的過錶達可能通過靶嚮下調EphA2的錶達從而抑製OSCC的增殖。
목적：탐토miR-141대구강린상세포암（ OSCC） CAL27세포증식활성적영향급기여EphA2적파향관계。방법구건pre-miR-141、EphA2야생형（EphA2-WT）화돌변형（EphA2-MT）진핵표체재체，채용실시정량PCR（qRT-PCR）검측pre-miR-141적전염효솔。용MTT검측세포증식활성，용쌍형광소매보고기인검측、qRT-PCR화Western blot험증miR-141여EphA2적파향관계。결과 pcDNATM6.2-GW-pre-miR-141、pmirGLO-EphA2-WT화pmirGLO-E-phA2-MT표체재체구건성공，전염pre-miR-141적CAL27세포miR-141적표체현저승고（ P＜0.001）。 miR-141과표체능구명현억제CAL27세포적증식（ P＜0.05），공전염pre-miR-141화EphA2-WT적CAL 27세포기형광활성현저하강（P＜0.001），전염pre-miR-141적CAL27세포기EphA2적표체요현저저우전염공질립조（P＜0.001），miR-141적과표체능구현저억제EphA2적단백표체수평。결론 miR-141적과표체가능통과파향하조EphA2적표체종이억제OSCC적증식。
Objective To assess the effect of miR-141 on proliferation of human oral squamous cell carcinoma and target relationship between miR-141 and EphA2 .Methods pcDNATM6.2-GW-pre-miR-141 was constructed and identified by qRT-PCR.EphA2-WT and EphA2-MT sequences were respectively cloned into pmirGLO plasmid . The potential proliferation function of miR-141 on CAL27 cells was analyzed by MTT .The target relationship be-tween miR-141 and EphA2 was identified by Dual-Luciferase Assay System , qRT-PCR and Western blot .Results We constructed successfully the recombinant plasmids , including pcDNATM6.2-GW-pre-miR-141, pmirGLO-E-phA2-WT and pmirGLO-EphA2-MT, and the transfection efficiency of pre-miR-141 was increased in CAL27 cells compared to control group(P<0.001).miR-141 could suppress the proliferation of CAL27 cells(P<0.05). Furthermore, a significant reduction of luciferase activities of CAL27 cells co-transfected with pre-miR-141 and EphA2-WT(P<0.001).The mRNA(P<0.001) and protein expression levels of EphA2 were decreased in CAL27 cells transfected with pre-miR-141 .Conclusions Overexpression of miR-141 may suppress cell prolifera-tion by targeting at EphA2 in CAL27 cells.
