CAJ | 학술논문

目的通过Smoothened (Smo)基因位点阻断Hh信号通路抑制食管癌细胞增殖,并诱导其凋亡.方法运用Smo小干扰RNA(siRNA)转染食管癌EC9706细胞,采用逆转录-聚合酶链反应(RT-PCR)和Western blot技术检测各组细胞Smo和Glil mRNA及蛋白表达,并以噻唑蓝(MTT)法和流式细胞术检测SmosiRNA对细胞增殖及凋亡的影响.结果 (1)Smo siRNA转染细胞24、48和72 h后,Smo mRNA相对表达量分别为4.20±1.10、2.80±1.10和1.00±0.71；GlilmRNA相对表达量分别为4.60±1.34、3.00±0.71和1.20 ±1.10,与各对照组比较均明显降低(P＜0.05)；(2) Smo siRNA转染细胞72 h后,Smo和Glil蛋白相对表达水平分别为0.330±0.016和0.324±0.021,与各对照组比较均明显下降(P＜0.05)；(3)SmosiRNA转染细胞72 h后,细胞生长抑制率为(88.06±7.56)％,明显高于各对照组,并随转染时间延长抑制率升高(P＜0.05)；(4)SmosiRNA转染细胞72h后,早期凋亡细胞所占的比例均明显增高.结论 Smo基因具有调控食管癌细胞增殖和凋亡的作用.
목적 통과Smoothened (Smo)기인위점조단Hh신호통로억제식관암세포증식,병유도기조망.방법 운용Smo소간우RNA(siRNA)전염식관암EC9706세포,채용역전록-취합매련반응(RT-PCR)화Western blot기술검측각조세포Smo화Glil mRNA급단백표체,병이새서람(MTT)법화류식세포술검측SmosiRNA대세포증식급조망적영향.결과 (1)Smo siRNA전염세포24、48화72 h후,Smo mRNA상대표체량분별위4.20±1.10、2.80±1.10화1.00±0.71；GlilmRNA상대표체량분별위4.60±1.34、3.00±0.71화1.20 ±1.10,여각대조조비교균명현강저(P＜0.05)；(2) Smo siRNA전염세포72 h후,Smo화Glil단백상대표체수평분별위0.330±0.016화0.324±0.021,여각대조조비교균명현하강(P＜0.05)；(3)SmosiRNA전염세포72 h후,세포생장억제솔위(88.06±7.56)％,명현고우각대조조,병수전염시간연장억제솔승고(P＜0.05)；(4)SmosiRNA전염세포72h후,조기조망세포소점적비례균명현증고.결론 Smo기인구유조공식관암세포증식화조망적작용.
Objective To investigate the proliferation and apoptosis of blocking Hh signaling on Smo gene locus in esophageal cancer cell.Methods The small interfering RNA (siRNA) was transfected into esophageal squamous carcinoma cell EC9706.reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the levels of Smo and Gli1 mRNA and protein.Methyl thiazol tetrazolium (MTT) assay and flow cytometry were used to detect the cell proliferation and apoptosis.Results (1) Compared with the control groups,after transfectitoa of Smo siRNA for 24,48 and 72 h,the levels of Smo mRNA were 4.20 ± 1.10,2.80 ± 1.10 and 1.00 ± 0.71,Gli1 mRNA 4.60 ± 1.34,3.00 ±0.71 and 1.20 ± 1.10,which were knocked down significantly (P ＜0.05).(2) After transfection of Smo siRNA for 72 h,the levels of Smo and Gli protein were 0.330 ±0.016 and 0.324 ±0.021 respectively,which were significantly lower than the control groups (P ＜ 0.05).(3) The inhibition rate of cell proliferation was (88.06 ± 7.56) ％ after transfection of 72 h,which was significantly inhibited compared with the control groups.(4) The number of apoptosis cells was also greatly increased after Smo siRNA transfection.Conclusion Smo gene may play an important role in the proliferation and apoptosis of esophageal cancer cells.