UNASSIGNED: To accomplish this objective, Whole Exome Sequencing (WES) data of 400 patients with unselected BC was used to filter variants located in the region of interest of BRCA1 gene, finding two of them (c.*36C>G and c.*369_373del). miRGate and miRanda in silico tools were used to predict microRNA (miRNA) interaction.
UNASSIGNED: The two variants (c.*36C>G, c.*369_373del) were predicted to affect miRNA interaction. After cloning of BRCA1 3\'UTR into pMIR-Report vector, the construct was transfected into two BC cell lines (MDA-MB-231 and MCF-7), and the variant c.*36C>G evidenced overexpression of reporter gene luciferase, showing that the transcript was not being degraded by the miRNA in MDA-MB-231 cells.
UNASSIGNED: The variant seems to protect against Triple Negative BC probably due to the expression level of miRNA in this particular cell line (MDA-MB-231). This is consistent with the clinical history of the patients who harbor BC Hormone Receptors positive (HR+).
■为了实现这一目标,全外显子组测序(WES)的400名患者的未选择的BC被用来过滤变异位于感兴趣的BRCA1基因的区域,找到其中两个(c。*36C>G和C.*369_373del)。miRGate和miRanda在计算机工具中用于预测microRNA(miRNA)相互作用。
■两种变体(c。*36C>G,c.*369_373del)预测会影响miRNA相互作用。在将BRCA13\'UTR克隆到pMIR-Report载体中之后,将构建体转染入两种BC细胞系(MDA-MB-231和MCF-7),变体c.*36C>G证明了报告基因荧光素酶的过表达,显示转录本在MDA-MB-231细胞中没有被miRNA降解。
■该变体似乎对三阴性BC具有保护作用,这可能是由于miRNA在该特定细胞系(MDA-MB-231)中的表达水平。这与携带BC激素受体阳性(HR+)的患者的临床病史一致。