贵州医科大学学报

2017, v.42;No.204(09) 1003-1009

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

梯度浓度顺铂处理对6种肿瘤细胞中CASP9的mRNA选择性剪接的影响
Effect of Gradient Concentration of Cisplatin on Alternative Splicing of CASP9 mRNA in 6 Tumor Cells and Its Mechanism

杨勤兴;马春霞;陆成龙;杨艳波;杨梦莉;刘舒媛;李明阳;杨渊;史荔;俞建昆;
YANG Qinxing;MA Chunxia;LU Chenglong;YANG yanbo;YANG Mengli;LIU Shuyuan;LI Mingyang;YANG Yuan;SHI Li;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;

摘要(Abstract):

目的:分析顺铂处理后多种肿瘤细胞中CASP9基因mRNA选择性剪接异构体的表达及其变化规律。方法:用梯度浓度的顺铂处理肺癌细胞(A549、H1299)、人胚肾细胞(293FT)、宫颈癌细胞(SiHa、CaSki、C33A)6种细胞,提取各细胞的总RNA,用半定量RT-PCR和琼脂糖凝胶电泳及胶图像分析软件检测分析CASP9基因mRNA异构体的组成及其比例和相对表达水平的变化。结果:6种细胞中,CASP9f∶r1引物检测到的CASP9a和c(NR-102732)两种异构体,但随着顺铂浓度的增加,两种异构体的比例在6种细胞中均无变化;而CASP9f∶r2引物可检测到的异构体CASP9a和b、a/b比例在A549、H1299、SiHa、C33A、Ca Ski中呈逐渐增加趋势,而在293FT细胞中则呈相反趋势;在A549顺铂抗性株中,顺铂处理前后CASP9a/b的比例显著差异变化明显减弱。结论:在顺铂处理下,检测的5种细胞中的Casp9是通过增加casp9a/b的比例来应对,仅293FT例外;顺铂抗性A549细胞株在顺铂处理后casp9a/b的比例比例与细胞凋亡和抗性有关,提示调节casp9a/b的比例是治疗癌症的可能的有效途径之一。
Objective: To explore the expression of alternative splicing isoforms of CASP9 mRNA in 6tumor cells treated with gradient concentration of cisplatin. Methods: Lung cancer( A549 and H1299) and cervical cancer( SiHa,Ca Ski and C33A) and 293 FT cell lines were treated by gradient dose of cisplatin. Total RNAs of the control cells and the treated cells were extracted 24 h after the treatment. The composition,proportion and relative level of mRNA isoforms in CASP9 gene were detected by semiquantitative RT-PCR,agarose gel electrophoresis and image analysis software. Results:In 6 cell lines,mRNA isoforms CASP9 a and c( NR-102732) were detected by using primers CASP9f∶r1,but their ratio didn't changed when the concentration of cisplatin increased. When using primers CASP9f∶ r2,mRNA isoforms CASP9 a and b could be detected. Except 293 FT,in other 5 cell lines,the ratio of CASP9a/b increased with rise of cisplatin concentration. In 293 FT,the ratio of CASP9a/b decreased. In cisplatin resistant cell line,A549-CIS-R,the obvious increase of CASP9a/b ratio was abated markedly after cisplatin treatment compared with normal cell line. Conclusion: Except 293 FT,in other 5 cell lines,cell regulated pre-mRNA alternative splicing of CASP9 in response to DNA damage induced by cisplatin through up-regulating the ratio of CASP9 a/b,but not down-regulating mRNA isoforms c,d. Combined with the result from cisplatin resistant cell line,A549-CIS-R,it is suggested that the ratio of CASP9 a/b can influence cell apoptosis and cisplatin resistance,and regulating ratio of CASP9 a/b is a considerable way for cancer treatment.

关键词(KeyWords): 顺铂;CASP9;mRNA选择性剪接;mRNA异构体;细胞凋亡
cisplatin;CASP9;mRNA alternative splicing;mRNA isoform;cell apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(31570918);; 中国医学科学院医学与健康科技创新工程重大协同创新项目(2016-12M-2-001)

作者(Author): 杨勤兴;马春霞;陆成龙;杨艳波;杨梦莉;刘舒媛;李明阳;杨渊;史荔;俞建昆;
YANG Qinxing;MA Chunxia;LU Chenglong;YANG yanbo;YANG Mengli;LIU Shuyuan;LI Mingyang;YANG Yuan;SHI Li;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;

Email:

DOI: 10.19367/j.cnki.1000-2707.2017.09.003

参考文献(References):

文章评论(Comment):

序号(No.) 时间(Time) 反馈人(User) 邮箱(Email) 标题(Title) 内容(Content)
反馈人(User) 邮箱地址(Email)
反馈标题(Title)
反馈内容(Content)
扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享