南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2015年
1期
88-92
,共5页
Stat1%c-Myc%宫颈癌%Hela细胞%顺铂%化学治疗
Stat1%c-Myc%宮頸癌%Hela細胞%順鉑%化學治療
Stat1%c-Myc%궁경암%Hela세포%순박%화학치료
Stat1%c-Myc%cervical carcinoma%HeLa cells%cisplatin%chemotherapy
目的：研究Stat1蛋白对宫颈癌Hela细胞生长、增殖及顺铂敏感性的影响及其作用机制。方法采用蛋白印迹法（Western blot）检测不同浓度DDP处理Hela细胞后Stat1蛋白表达差异；转染Stat1-siRNA后，通过Western blot验证干扰效果；应用四甲基偶氮唑蓝（MTT）法和5-溴脱氧尿嘧啶核苷（BrdU）观察转染Stat1-siRNA后Hela细胞对DDP敏感性变化，并检测Stat1相关细胞因子c-Myc的表达变化。结果随着DDP浓度的增加，Hela细胞中Stat1蛋白的表达量逐渐增加，DDP浓度为5 mg/L时，Stat1蛋白表达上调1.5倍，DDP浓度为10 mg/L时，Stat1蛋白表达上调近2倍；特异性Stat1-siRNA使Hela细胞中Stat1的表达下调70%，促进细胞生长和增殖；Stat1-siRNA可以降低DDP对Hela细胞生长和增殖的抑制效应，削弱DDP对c-Myc的抑制作用。结论 Hela细胞中c-Myc是STAT1发挥抑癌作用的靶点基因之一；STAT1/c-Myc通路介导了DDP抑制Hela细胞生长、增殖的作用；STAT1可增强Hela细胞对DDP的敏感性。
目的：研究Stat1蛋白對宮頸癌Hela細胞生長、增殖及順鉑敏感性的影響及其作用機製。方法採用蛋白印跡法（Western blot）檢測不同濃度DDP處理Hela細胞後Stat1蛋白錶達差異；轉染Stat1-siRNA後，通過Western blot驗證榦擾效果；應用四甲基偶氮唑藍（MTT）法和5-溴脫氧尿嘧啶覈苷（BrdU）觀察轉染Stat1-siRNA後Hela細胞對DDP敏感性變化，併檢測Stat1相關細胞因子c-Myc的錶達變化。結果隨著DDP濃度的增加，Hela細胞中Stat1蛋白的錶達量逐漸增加，DDP濃度為5 mg/L時，Stat1蛋白錶達上調1.5倍，DDP濃度為10 mg/L時，Stat1蛋白錶達上調近2倍；特異性Stat1-siRNA使Hela細胞中Stat1的錶達下調70%，促進細胞生長和增殖；Stat1-siRNA可以降低DDP對Hela細胞生長和增殖的抑製效應，削弱DDP對c-Myc的抑製作用。結論 Hela細胞中c-Myc是STAT1髮揮抑癌作用的靶點基因之一；STAT1/c-Myc通路介導瞭DDP抑製Hela細胞生長、增殖的作用；STAT1可增彊Hela細胞對DDP的敏感性。
목적：연구Stat1단백대궁경암Hela세포생장、증식급순박민감성적영향급기작용궤제。방법채용단백인적법（Western blot）검측불동농도DDP처리Hela세포후Stat1단백표체차이；전염Stat1-siRNA후，통과Western blot험증간우효과；응용사갑기우담서람（MTT）법화5-추탈양뇨밀정핵감（BrdU）관찰전염Stat1-siRNA후Hela세포대DDP민감성변화，병검측Stat1상관세포인자c-Myc적표체변화。결과수착DDP농도적증가，Hela세포중Stat1단백적표체량축점증가，DDP농도위5 mg/L시，Stat1단백표체상조1.5배，DDP농도위10 mg/L시，Stat1단백표체상조근2배；특이성Stat1-siRNA사Hela세포중Stat1적표체하조70%，촉진세포생장화증식；Stat1-siRNA가이강저DDP대Hela세포생장화증식적억제효응，삭약DDP대c-Myc적억제작용。결론 Hela세포중c-Myc시STAT1발휘억암작용적파점기인지일；STAT1/c-Myc통로개도료DDP억제Hela세포생장、증식적작용；STAT1가증강Hela세포대DDP적민감성。
Objective To investigate the role of Stat1 gene in the proliferation and chemotherapeutic sensitivity of cervical cancer HeLa cells. Methods The protein expression of Stat1 in the Hela cells exposed to gradient concentrations of cisplatin (DDP) was detected by Western blotting with or without small interfering RNA (siRNA)-mediated Stat1 gene silencing. The effect of Sata1 silencing on the sensitivity to DDP and cell proliferation of the cells was tested using MTT assay and BrdU assay, and the expression of c-Myc was detected by Western blotting in the cells treated with siRNA and DDP. Results The expression of Stat1 in Hela cells exposed to DDP increased with the DDP concentrations, reaching 1.5 folds of the baseline at a DDP concentration of 5 mg/L and 2 folds at 10 mg/L. Stat1-siRNA effectively reduced Stat1 expression in Hela cells, promoted the cell proliferation, and enhanced the expression of c-Myc;DDP inhibited the cell growth and down-regulated c-Myc expression. Stat1-siRNA rescued DDP-induced inhibition of cell growth and c-Myc down-regulation. Conclusion The expression of Stat1 is associated with DDP sensitivity in cervical cancer cells, and Stat1 silencing can increase the sensitivity to DDP and c-Myc expression of the cells.
