一种新的鳜肿瘤抑制因子p53剪接异构体克隆及其功能初探
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- 英文论文题名：Cloning and function of a new kind of splicing isoform for tumor suppressor protein p53 in mandarin fish Siniperca chuatsi
- 论文作者：郭慧芝 ; 付小哲 ; 李宁求
- 英文论文作者：Huizhi Guo ; Xiaozhe Fu ; Ningqiu Li ; Pearl River Fishery Research Institute ; Chinese Academy of Fishery Sciences ; Key Laboratory of Fishery Drug Development ; Ministry of Agriculture ; Key Laboratory of Aquatic Animal Immune Technology ; Guangdong Provinces
- 年：2016
- 作者机构：中国水产科学研究院珠江水产研究所农业部渔用药物创制重点实验室广东省水产动物免疫技术重点实验室;
- 论文关键词：p53 ; 剪接异构体 ; SiRNA ; 细胞周期
- 英文论文关键词：p53 ; splicing isform ; SiRNA ; cell cycle
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：561k
- 原文格式：D
- 会议级别：全国
摘要
p53是一种重要的肿瘤抑制因子,具有调节细胞周期,控制细胞死亡,诱导细胞凋亡,参与细胞代谢和抗病毒等功能。本研究采用RACE技术获得了鳜p53的一种新剪接异构体(Sc-p53a)的cDNA全长,编码254个AA。相比鳜p53的cDNA全长少了5个外显子,并保留了第六个内含子。荧光定量PCR检测健康鳜的Sc-p53a在各个组织中呈组成型表达,主要表达于鳃、血细胞和后肾中。经鳜传染性脾肾坏死病毒(ISKNV)感染后,Sc-p53a在鳜脑组织细胞系(CPB)表达呈现双相性上升的变化,在感染3小时表达显著上调,随后下降,感染96小时表达再次显著上调;ISKNV感染鳜鱼后在脾脏组织中的表达情况与感染细胞情况相似。采用RNA干扰技术,针对可变剪接区域构建SiRNA载体(SiRNA-p53a),转染CPB细胞能显著下调Sc-p53a的表达。转染细胞经过约28天的生长后,细胞死亡,表明Sc-p53a可能与细胞周期的调控有关。该研究为p53的剪接异构体提供了新的资料,为深入了解p53的调节细胞周期和抗病毒等功能奠定了基础。
P53 is an important tumor suppressor,which is regulate the cell cycle, control cell death,inducing cell apoptosis, and participates in cell metabolism and antivirus. In the present study, a new splicing isoform for Siniperca chuatsi(Sc-p53a), the full length c DNA with coded 254 AA has been cloned by RACE, which less five exons than Siniperca chuatsi p53(Sc-p53) and keep the sixth introns. Quantitative real-time PCR assays revealed that Sc-p53a was expressed in all tissues examined, and it was most abundant in the gill, hemocyte and hind-kidney. The mRNA expression of Sc-p53a was significantly up-regulated in the Chinese perch(CPB) cell line and mandarin fish after infection with infectious kidney and spleen necrosis virus(ISKNV). The results showed a biphasic expression pattern of Sc-p53a in CPB and spleen at 3 and 96 hours.Using RNA interference technology, SiRNA vector(SiRNA-p53a) is constructed in alternative splicing area. CPB cell is transfected in SiRNA-p53a, which can significantly lower the expression of Sc-p53a. After 28 days the transfected cells were died. The results suggest that Sc-p53a may be associated with the regulation of cell cycle. The study has provided the new material for splicing isomer of p53and lay the foundation for cell cycle and antiviral function.
P53 is an important tumor suppressor,which is regulate the cell cycle, control cell death,inducing cell apoptosis, and participates in cell metabolism and antivirus. In the present study, a new splicing isoform for Siniperca chuatsi(Sc-p53a), the full length c DNA with coded 254 AA has been cloned by RACE, which less five exons than Siniperca chuatsi p53(Sc-p53) and keep the sixth introns. Quantitative real-time PCR assays revealed that Sc-p53a was expressed in all tissues examined, and it was most abundant in the gill, hemocyte and hind-kidney. The mRNA expression of Sc-p53a was significantly up-regulated in the Chinese perch(CPB) cell line and mandarin fish after infection with infectious kidney and spleen necrosis virus(ISKNV). The results showed a biphasic expression pattern of Sc-p53a in CPB and spleen at 3 and 96 hours.Using RNA interference technology, SiRNA vector(SiRNA-p53a) is constructed in alternative splicing area. CPB cell is transfected in SiRNA-p53a, which can significantly lower the expression of Sc-p53a. After 28 days the transfected cells were died. The results suggest that Sc-p53a may be associated with the regulation of cell cycle. The study has provided the new material for splicing isomer of p53and lay the foundation for cell cycle and antiviral function.
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