Brahami Nabila, Subramaniam Selvakumar, Al-Ddafari Moudjahed Saleh, Elkaim Cecile, Harmand Pierre-Olivier, Sari Badr-Eddine, Lefranc Gérard, Aribi Mourad
Laboratory of Applied Molecular Biology and Immunology, University of Tlemcen, Imama-Mansourah, Rocade # 2, PO Box: 262, Tlemcen, 13000, Algeria.
UMR U866 INSERM, University of Burgundy, 21000, Dijon, France.
J Negat Results Biomed. 2017 Mar 20;16(1):9. doi: 10.1186/s12952-017-0072-5.
We aimed to search for mutations in the germline and somatic DNA of the TEK gene and to analyze the expression level of Src and phospho-Src (p-Src) in tumor and healthy tissues from patients with facial cutaneo-mucosal venous malformations (VMCM). Eligible patients from twelve families and thirty healthy controls were recruited respectively at the Departments of Stomatology and Oral Surgery, and Transfusion Medicine of Tlemcen University Medical Centre. Immunoblot analyses of Src and p-Src were performed after direct DNA sequencing. No somatic or germline mutations were found in all the 23 exons and their 5' and 3' intronic flanking regions, except for one case in which a c.3025+20-3025+22 del mutation was highlighted at the intron 15, both in the germline and somatic DNA. Additionally, elevated expression levels of Src and p-Src were observed only in the patient with such mutation. However, when normalized to β-actin, the overall relative expression levels of both Src and p-Src were significantly increased in VMCM tissues when compared to healthy tissues (for both comparisons, p <0.001). In conclusion, we confirm the outcomes of our previous work suggesting that VMCM can develop independently of mutation of the TEK gene. Additionally, the results for Src activity are of particular interest in the context of specific targeted therapies and biological diagnosis. Nevertheless, such a conclusion should be confirmed through a mechanistic study and/or in a satisfactory number of patients.
我们旨在寻找TEK基因种系和体细胞DNA中的突变,并分析面部皮肤黏膜静脉畸形(VMCM)患者肿瘤组织和健康组织中Src和磷酸化Src(p-Src)的表达水平。分别从特莱姆森大学医学中心的口腔颌面外科和输血医学科招募了来自12个家庭的符合条件的患者和30名健康对照。在直接DNA测序后进行Src和p-Src的免疫印迹分析。在所有23个外显子及其5'和3'内含子侧翼区域均未发现体细胞或种系突变,只有1例在第15内含子中突出显示了一个c.3025+20-3025+22缺失突变,该突变在种系和体细胞DNA中均存在。此外,仅在有此类突变的患者中观察到Src和p-Src的表达水平升高。然而,与β-肌动蛋白标准化后,与健康组织相比,VMCM组织中Src和p-Src的总体相对表达水平均显著升高(两项比较均p<0.001)。总之,我们证实了我们之前工作的结果,表明VMCM可以独立于TEK基因突变而发生。此外,可以认为Src活性的结果在特定靶向治疗和生物学诊断方面具有特殊意义。然而,这一结论应通过机制研究和/或在足够数量的患者中得到证实。