位置:成果数据库 > 期刊 > 期刊详情页
抑制差减杂交方法筛选辐射诱导细胞恶性转化模型中的差异表达基因
  • ISSN号:0258-879X
  • 期刊名称:《第二军医大学学报》
  • 时间:0
  • 分类:R827.31[医药卫生—临床医学]
  • 作者机构:[1]第二军医大学海军医学系放射医学教研室,上海200433
  • 相关基金:国家自然科学基金(30370359)
中文摘要:

目的:筛选辐射诱导的支气管上皮细胞恶性转化模型中的差异表达基因。方法:应用抑制差减杂交方法(SSH)构建辐射诱导的细胞转化模型差异表达基因的cDNA文库。对差减文库进行PCR筛选,对得到的差异片段进行测序及BLAST分析;对部分筛选出来的差异表达基因使用荧光定量PCR方法检测并确认其变化;将新的序列表达标签(EST)登录到Gen-Bank上。结果:在80个进行测序的克隆中,得到确定序列的共73个。在转化细胞中表达下降的序列41条,BLAST比较分析结果:得到已知序列6条;未知基因的EST序列20条;空载体序列7条;8条为重复测定序列。在转化细胞中表达上升的序列32条,BLAST分析:已知序列14条;未知基因的EST序列9条;重复测定的序列9条。对其中的部分基因改变进行荧光定量PCR检测,结果表明辐射转化组中MY06、HACE1、ZNF143、HNRPH1的表达量明显增加(与对照组相比,转化组中的mRNA分别增加了3.49、29.38、12.99、5.00倍);而PCBP2、RPL15、TCERG1的表达量下降(与对照组相比,转化组中的mRNA分别减少了1.89、48.77、11.95倍)。将得到的29个未知序列登录到GenBank,序列ID:EB643220~EB643248。结论:利用抑制差减杂交方法成功建立了恶性转化细胞模型差异表达cDNA文库,包含大量功能未知的新基因;ZNF143表达增加,其与细胞的增殖与分裂有关,TCERG1作为转录辅助激活因子在mRNA转录和后期的修饰过程中起到重要的作用,PCBP2是一个Polyc连接蛋白,具有蛋白翻译调节功能,这些基因在辐射致癌中尚无研究报道。

英文摘要:

Objective:To screen for the differentially expressed genes during irradiation induced malignant transformation of human bronchus epithelium cells (BEAS-2B). Methods: Suppression subtraction hybridization (SSH) was used to construct a subtracted cDNA library of differentially expressed genes during irradiation-induced malignant transformation of BEAS 2B cells. Then the subtracted library was screened by PCR and the differential fragments were sequenced and analyzed with BLAST. Fluorescent real time quantitative PCR was used to investigate some of the differentially expressed genes. The new EST was registered in GenBank. Results: Then 40 clones were chosen to be sequenced from the library of increased expression and decreased expression respectively according to the length of insertion element. Totally 73 sequences were obtained from the 80 sequenced clones. Forty one sequences were decreased in the transformed cells; BLAST analysis indicated that there were 6 known sequences,20 unknown sequences, 7 void sequences and 8 repeated sequences. Thirty-two sequences were increased in the transformed cells; Blast analysis indicated that there were 14 known sequences,9 unknown sequences,and 9 repeated sequences. Fluorescent quantitative PCR revealed that, compared with control group, the expression of MY06, HACE1, ZNF143, and HNRPH1 were significantly increased in the radiation transforming group, with their mRNAs increased by 3.49,29.38,12.99 and 5.00 folds, respectively. Compared with control group, the expression of PCBP2, RPL15, and TCERG1 in the radiation transforming group was significantly decreased, with their mRNAs decreased by 1.89,48.77 and 11.95 folds, respectively. The 29 unknown sequences were registered in the GenBank (ID: EB643220-EB643248). Conclusion: The cDNA library has been successfully established for malignant transformation cellular model by suppression subtractive hybridization;the library includes a number of unknown genes. The increased gene ZNF143 is associated with cell pr

同期刊论文项目
同项目期刊论文
期刊信息
  • 《第二军医大学学报》
  • 北大核心期刊(2011版)
  • 主管单位:第二军医大学
  • 主办单位:第二军医大学
  • 主编:吴孟超
  • 地址:上海市翔殷路800号
  • 邮编:200433
  • 邮箱:bxue@smmu.edu.cn
  • 电话:021-81870791
  • 国际标准刊号:ISSN:0258-879X
  • 国内统一刊号:ISSN:31-1001/R
  • 邮发代号:4-373
  • 获奖情况:
  • 2008年被评为首批"中国精品科技期刊"2008年获第二...,2004年获第四届全军医学期刊质量评比优秀奖,2002年获第二届国家期刊奖百种重点期刊奖,2000年获首届《CAJ-CD规范》执行评优活动执行优秀奖,1999年获上海高校优秀自然科学学报评比一等奖,1999年获全国高校自然科学学报及教育部优秀科技期...,1997年获上海科技期刊评比二等奖
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,荷兰文摘与引文数据库,荷兰医学文摘,美国剑桥科学文摘,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:30859