中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
3期
187-192
,共6页
郭跃辉%高丰厚%时婧%袁海花%姜斌
郭躍輝%高豐厚%時婧%袁海花%薑斌
곽약휘%고봉후%시청%원해화%강빈
肺肿瘤%微小RNA145%表皮生长因子受体%细胞外信号调节激酶1/2%信号传导
肺腫瘤%微小RNA145%錶皮生長因子受體%細胞外信號調節激酶1/2%信號傳導
폐종류%미소RNA145%표피생장인자수체%세포외신호조절격매1/2%신호전도
Lung neoplasms%MiRNA-145%Epidermal growth factor receptor%ERK1/2%Signal transduction
目的 探讨肺癌细胞中表皮生长因子受体(EGFR)的活化与微小RNA145(miR-145)下调之间的关系及其分子机制.方法 选择正常肺上皮细胞BEAS-2B、EGFR野生型和突变型肺癌细胞A549、H292、H1650和H1975,采用实时荧光定量PCR和Western blot法检测各细胞中miR-145的表达水平和EGFR的活化水平,并分析二者的相关性.分别以EGF、特异性EGFR酪氨酸激酶抑制剂AG1478或ERK1/2抑制剂U0126处理H1975、A549和BEAS-2B细胞,观察各细胞中miR-145的表达.结果 肺癌细胞中EGFR的活化与miR-145的表达呈显著负相关(r=-0.926,P=0.024);EGF可下调miR-145的表达,以BEAS-2B和A549细胞内miR-145的表达下调最为明显,分别下调53.0％(t=30.993,P=0.001)和42.6％(t=14.326,P=0.005);AG1478可恢复EGFR活化诱导miR-145的下调,经AG1478处理后,H1975细胞内miR-145的表达上调67.5％(t=8.269,P=0.014).EGFR活化后可激活下游信号蛋白分子ERK1/2,U0126可逆转EGFR活化所诱导的miR-145的下调.结论 在肺癌细胞中,EGFR通过ERK1/2信号分子下调miR-145的表达.
目的 探討肺癌細胞中錶皮生長因子受體(EGFR)的活化與微小RNA145(miR-145)下調之間的關繫及其分子機製.方法 選擇正常肺上皮細胞BEAS-2B、EGFR野生型和突變型肺癌細胞A549、H292、H1650和H1975,採用實時熒光定量PCR和Western blot法檢測各細胞中miR-145的錶達水平和EGFR的活化水平,併分析二者的相關性.分彆以EGF、特異性EGFR酪氨痠激酶抑製劑AG1478或ERK1/2抑製劑U0126處理H1975、A549和BEAS-2B細胞,觀察各細胞中miR-145的錶達.結果 肺癌細胞中EGFR的活化與miR-145的錶達呈顯著負相關(r=-0.926,P=0.024);EGF可下調miR-145的錶達,以BEAS-2B和A549細胞內miR-145的錶達下調最為明顯,分彆下調53.0％(t=30.993,P=0.001)和42.6％(t=14.326,P=0.005);AG1478可恢複EGFR活化誘導miR-145的下調,經AG1478處理後,H1975細胞內miR-145的錶達上調67.5％(t=8.269,P=0.014).EGFR活化後可激活下遊信號蛋白分子ERK1/2,U0126可逆轉EGFR活化所誘導的miR-145的下調.結論 在肺癌細胞中,EGFR通過ERK1/2信號分子下調miR-145的錶達.
목적 탐토폐암세포중표피생장인자수체(EGFR)적활화여미소RNA145(miR-145)하조지간적관계급기분자궤제.방법 선택정상폐상피세포BEAS-2B、EGFR야생형화돌변형폐암세포A549、H292、H1650화H1975,채용실시형광정량PCR화Western blot법검측각세포중miR-145적표체수평화EGFR적활화수평,병분석이자적상관성.분별이EGF、특이성EGFR락안산격매억제제AG1478혹ERK1/2억제제U0126처리H1975、A549화BEAS-2B세포,관찰각세포중miR-145적표체.결과 폐암세포중EGFR적활화여miR-145적표체정현저부상관(r=-0.926,P=0.024);EGF가하조miR-145적표체,이BEAS-2B화A549세포내miR-145적표체하조최위명현,분별하조53.0％(t=30.993,P=0.001)화42.6％(t=14.326,P=0.005);AG1478가회복EGFR활화유도miR-145적하조,경AG1478처리후,H1975세포내miR-145적표체상조67.5％(t=8.269,P=0.014).EGFR활화후가격활하유신호단백분자ERK1/2,U0126가역전EGFR활화소유도적miR-145적하조.결론 재폐암세포중,EGFR통과ERK1/2신호분자하조miR-145적표체.
Objective To investigate the relationship between EGFR activation and down-regulation of miRNA-145 in lung cancer.Methods Normal human lung epithelia cell line (BEAS-2B),human lung adenocarcinoma cell lines with wild-type EGFR (A549 and H292) and human lung adenocarcinoma cell lines with EGFR mutation (H1975 and H1650) were chosen in this study.The levels of miRNA-145 and pEGFR were determined by quantitative real-time PCR (qRT-PCR) and Western blotting,respectively,and the relationship between p-EGFR and miRNA-145 levels was analyzed.The miRNA-145 levels were determined by qRT-PCR after activating EGFR with EGF or blocking EGFR signal pathway with AG1478.In addition,ERK1/2 inhibitor U0126 was used to inhibit ERK1/2 activation and then the expression of miRNA-145 was detected.Results The miRNA-145 levels were closely negatively related with p-EGFR in lung cancer cells (r =-0.926,P =0.024).EGF down-regulated miRNA-145 expression,particularly in BEAS-2B cells (53.0％ ; t =30.993,P =O.001) and A549 ceils (42.6％ ; t =14.326,P =O.005).The miRNA-145 was up-regulated after inhibiting p-EGFR with AG1478,and significantly enhanced by 67.5％ in H1975 cells when treated with AG1478 (t =8.269,P =0.014).The ERK1/2 signal pathway was activated by p-EGFR.U0126 restored the miRNA-145 down-regulation induced by EGFR-activation in lung cancer cells.Conclusion The activation of EGFR down-regulates miRNA-145 expression through ERK1/2 in lung cancer cells.
