LMP1羧基端活化区3对鼻咽癌干细胞SP18迁移与侵袭的影响
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南华大学肿瘤研究所

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湖南省自然科学衡阳联合基金(12JJ9033),湖南省自然科学基金(13JJ3079),湖南省科技厅项目(2013SK3118,2014SK3081),湖南省高校创新平台基金(10K052,12K094,13K083),湖南省教育厅课题(11C1112,12C0340),湖南省卫生厅课题(B2013-048),湖南省研究生科研创新项目(CX2010B380),南华大学研究生创新项目(29),衡阳市科技局项目(2013KJ12,2013KJ28),邵阳市科技局重点项目(2013SK13),湖南省重点学科建设项目(2011-76)资助


The Effect of Migration and Invasion by Carboxy Terminal Activating Region 3 of LMP1 in Nasopharyngeal Carcinoma Stem Cell
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Cancer Research Institute, University of Southern China

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This work was supported by grants from the Hunan Provincial Natural Science Committee and Hengyang City Government Unification Foundation of China (12JJ9033), the Hunan Provincial Natural Science Foundation of China (13JJ3079), the Hunan Provincial Science and Technology Department Foundation of China (2013SK3118, 2014SK3081), the Hunan Provincial Groundbreaking Platform Open Fund of University of China (10K052, 12K094, 13K083), the Hunan Provincial Education Department Foundation of China (11C1112, 12C0340), the Hunan Provincial Health Brueau Department Foundation of China (B2013-048), the Hunan Provincial Postgraduate Groundbreaking Scientific Research Foundation of China (CX2010B380), the Postgraduate Groundbreaking Scientific Research Foundation of University of Southern China of China (29), the Hunan Provincial Hengyang city Science and technology Department Key Foundation of China (2013KJ12, 2013KJ28),...

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    摘要:

    为了观察潜伏性膜蛋白1(LMP1)羧基端活性区3(CTAR3)对鼻咽癌干细胞SP18迁移与侵袭的影响,本研究通过建立稳定表达LMP1及CTAR3突变型LMP1(LMP1△252-351)的SP18细胞系(即SP18-LMP1和SP18-LMP1△252-351),观察LMP1-CTAR3缺失突变后对SP18细胞增殖、迁移与侵袭的影响.采用基因芯片分析SP18-LMP1和SP18-LMP1△252-351间的差异表达基因,并验证基因的表达,用生物信息学分析差异表达基因间的相互关系.结果显示:a.SP-LMP1△252-351细胞生长速度较SP-LMP1细胞明显变缓,克隆形成和迁移与侵袭能力降低(n=3,P < 0.05);b.鉴定出LMP1羧基端CTAR3影响SP18细胞迁移与侵袭的18个基因(其中表达上调基因13个,下调基因5个),经荧光定量PCR验证与基因芯片检测结果基本一致.c.13个差异基因间相互联系,网络节点联系最多的基因是FN1、MMP14、THBS1、ITGA2、IL1B和IL6基因.结果提示,LMP1羧基端CTAR3可能通过调节FN1、MMP14、THBS1、ITGA2、IL1B和IL6基因的表达,发挥其促鼻咽癌干细胞SP18细胞迁移与侵袭的功能.

    Abstract:

    To investigate mechanism of migration and invasion effected by carboxyl terminal activating region 3 (CTAR3) of Epstein-Barr virus encoded latent membrane protein 1(LMP1) in nasopharyngeal carcinoma stem cell SP18 cells, the SP18 cell of stable expressed LMP1 and deletion mutant type LMP1 (LMP1△232-351) were established (SP18-LMP1 and SP18-LMP1△252-351). The effect of LMP1-CTAR3 deletion mutant for cellular proliferation, migration and invasion were observed in SP18 cells. The differential expression genes between SP18-LMP1 and SP18-LMP1△252-351 cells were analyzed by cDNA chips, and the expression levels of partial identified genes were verified by fluorescent Real-time quantitative RT-PCR. The relation of differential expression genes were analyzed by bioinformatics. The results showed: (1) The ability of LMP1△232-351 promoting SP18 cell proliferation, migration and invasion was obviously decreased to compare with wild type LMP1(n=3, P < 0.05). (2) 18 genes, 13 up- and 5 down-regulated ones, of LMP1-CTAR3 mediated regulation with migration and invasion were identified from in SP18 cell lines. The differential expression of partial identified genes was similar with cDNA chips separated ones and confirmed by fluorescent Real-time quantitative RT-PCR. (3) 13 differential expression genes can be cross-talk, and among FN1, MMP14, ITGA2, THBS1, IL1B and IL6 genes were frequently correlation. These results suggested that LMP1-CTAR3 probably regulates the expression of FN1, MMP14, ITGA2, THBS1, IL1B and IL6 genes to induce and promote migration and invasion of nasopharyngeal carcinoma stem cell SP18 cell.

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肖娟,张志伟,伍石华,刘重元,唐薇,赵强,雷明生,杨代水. LMP1羧基端活化区3对鼻咽癌干细胞SP18迁移与侵袭的影响[J].生物化学与生物物理进展,2015,42(1):65-72

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  • 收稿日期:2014-06-19
  • 最后修改日期:2014-08-14
  • 接受日期:2014-08-20
  • 在线发布日期: 2015-01-19
  • 出版日期: 2015-01-20