Genomic and proteomic profiling I: Leiomyomas in African Americans and Caucasians
Genomic and proteomic profiling I: Leiomyomas in African Americans and Caucasians
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DOI：
10.1186/1477-7827-5-34
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发表时间：
2007-08-23
影响因子：
4.4
通讯作者：
Chegini, Nasser
中科院分区：
文献类型：
--
作者：
Pan, Qun;Luo, Xiaoping;Chegini, Nasser
背景资料：临床观察表明，平滑肌瘤更频繁地发生在非洲裔美国人相比，其他种族群体的病因不明。为了确定差异的分子基础，我们比较了A型平滑肌瘤。方法：微阵列，实时PCR，2D-PAGE，质谱，蛋白质印迹和免疫组化。结果：使用Affyphase U133 A阵列和基于P排序的分析（P < 0.01），1470个基因被鉴定为与子宫肌层相比在平滑肌瘤中差异表达，而不考虑种族。其中，268个基因在A型平滑肌瘤中过表达（177个基因）或低表达（91个基因），基于P < 0.01，随后进行2倍截止选择。美国人与高加索人相比。其中，E2 F1、RUNX 3、EGR 3、TBPIP、ECM 2、ESM 1、THBS 1、GAS 1、ADAM 17、CST 6、CST 7、FBLN 5、ICAM 2、EDN 1和COL 18的表达通过实时荧光PCR低密度芯片验证。2D PAGE结合图像分析鉴定了332个蛋白质点，其中31个蛋白质点的密度/体积在平滑肌瘤中的变化大于或等于子宫肌层的1.5倍。A.平滑肌瘤中有34个蛋白质斑点密度/体积变化大于或等于1.5倍（26个增加，8个减少）。美国人与高加索人相比。对15个蛋白质点的串联质谱分析鉴定了几种蛋白质，其转录本也通过微阵列鉴定，包括14-3-3 β和mimecan，其表达通过蛋白质印迹和免疫组织化学确认。这些发现意味着许多基因和蛋白质的水平而不是种族特异性表达可能解释了平滑肌瘤和可能的子宫肌层之间的差异，以.美国人和高加索人。需要使用更大样本量的进一步研究来证实这些发现。
Background: Clinical observations indicate that leiomyomas occur more frequently in African Americans compared to other ethnic groups with unknown etiology. To identify the molecular basis for the difference we compared leiomyomas form A. Americans with Caucasians using genomic and proteomic strategies.Methods: Microarray, realtime PCR, 2D-PAGE, mass spectrometry, Western blotting and immunohistochemistry.Results: Using Affymetrix U133A array and analysis based on P ranking (P < 0.01) 1470 genes were identified as differentially expressed in leiomyomas compared to myometrium regardless of ethnicity. Of these, 268 genes were either over-expressed (177 genes) or under-expressed (91 genes) based on P < 0.01 followed by 2-fold cutoff selection in leiomyomas of A. Americans as compared to Caucasians. Among them, the expression E2F1, RUNX3, EGR3, TBPIP, ECM2, ESM1, THBS1, GAS1, ADAM17, CST6, CST7, FBLN5, ICAM2, EDN1 and COL18 was validated using realtime PCR low-density arrays. 2D PAGE coupled with image analysis identified 332 protein spots of which the density/volume of 31 varied by greater than or equal to 1.5 fold in leiomyomas as compared to myometrium. The density/volume of 34 protein-spots varied by greater than or equal to 1.5 fold (26 increased and 8 decreased) in leiomyomas of A. Americans as compared to Caucasians. Tandem mass spectrometric analysis of 15 protein spots identified several proteins whose transcripts were also identified by microarray, including 14-3-3 beta and mimecan, whose expression was confirmed using western blotting and immunohistochemistry.Conclusion: These findings imply that the level rather than the ethnic-specific expression of a number of genes and proteins may account for the difference between leiomyomas and possibly myometrium, in A. Americans and Caucasians. Further study using larger sample size is required to confirm these findings.