CAJ | 학술논문

目的建立人胃癌顺铂耐药细胞株,探讨其基因表达谱与顺铂耐药的相关性.方法采用浓度梯度递增法诱导胃癌SGC7901细胞株,建立对顺铂耐药的SGC7901/DDP细胞株.应用Affymetrix基因芯片HG-U133 Plus 2.0筛选SGC7901和SGC7901/DDP的耐药相关的差异基因.结果建立可耐受1.0 mg/L顺铂浓度的耐药株SGC7901/DDP,耐药指数22.85.应用电镜观察其形态学变化,细胞计数法绘制生长曲线,噻唑蓝(MTT)检测检测药物敏感性,流式细胞术分析细胞周期.利用基因芯片筛选得到在耐药细胞株和敏感株中表达差异大于10倍的基因共107条,其中JNK1和JNK2为与耐药相关的明显上调基因.Western blot证实JNK的磷酸化活性形式P-JNK在耐药株中表达明显升高.SP600125抑制P-JNK表达后耐药蛋白P-gp表达明显减低.结论高通量的基因芯片筛选得到大量有意义的与顺铂耐药相关的差异基因,P-JNK可成为逆转耐药的新的靶点.
목적 건립인위암순박내약세포주,탐토기기인표체보여순박내약적상관성.방법 채용농도제도체증법유도위암SGC7901세포주,건립대순박내약적SGC7901/DDP세포주.응용Affymetrix기인심편HG-U133 Plus 2.0사선SGC7901화SGC7901/DDP적내약상관적차이기인.결과 건립가내수1.0 mg/L순박농도적내약주SGC7901/DDP,내약지수22.85.응용전경관찰기형태학변화,세포계수법회제생장곡선,새서람(MTT)검측검측약물민감성,류식세포술분석세포주기.이용기인심편사선득도재내약세포주화민감주중표체차이대우10배적기인공107조,기중JNK1화JNK2위여내약상관적명현상조기인.Western blot증실JNK적린산화활성형식P-JNK재내약주중표체명현승고.SP600125억제P-JNK표체후내약단백P-gp표체명현감저.결론 고통량적기인심편사선득도대량유의의적여순박내약상관적차이기인,P-JNK가성위역전내약적신적파점.
Objective To establish cisplatin ( DDP)-resistant human gastric cancer cell line and investigate the relationship between differential gene profile and DDP resistance. Methods A DDP-resistant human gastric cancer cell line SGC7901/DDP was established by increasing dose of DDP up to 1.0 mg/L and the resistance index (RI) was 22.85. The muhidrug resistance (MDR) phenotype was tested by detecting cell morphology using electron microscopy,drug sensitivity using MTr array,and cell cycle using flow cytometry. Results 107 genes were identified,which were differentially expressed by more than 10 times between SGC7901 and SGC7901/DDP cell lines using Affymetrix gene chip HG-U133 Plus 2.0,among which JNK1 and JNK2 were significantly up-regulated in SGC7901/DDP cell line and the active phosphorylation type of JNK (P-JNK) was verified by Western blot. The expression of MDR protein P-gp was inhibited after the blocking of P-JNK by SP600125. Conclusion Numerous genes relating with DDPresistance were screened out by through-out gene chip and P-JNK could be the novel target gene for reversing MDR.