wnt6基因低表达对mda-mb-231细胞系生长增殖的影响word论文.docxVIP

对照组比较，实验组 WNT6 mRNA 及蛋白表达水平均分别下调近 50%和 40%，差异具有统计学意义(p0.05)；scRNA 组与未转染组比较差异无显著性(p0.05)。 MTT 法绘制生长曲线发现，与对照组比较，转染 48h 后，实验组细胞生长速度受 到明显抑制，细胞增殖率降低约 48%；转染 72h 后，实验组细胞增殖率降低约 49% ，差异具有统计学意义(p0.05)；scRNA 组与未转染组比较细胞增殖率差异 无显著性(p0.05)。细胞集落形成实验发现，实验组集落形成率较对照组降低，差 异有统计学意义(p0.05)；scRNA 组与未转染组相比，集落形成率无明显变化 (p0.05)；结果表明，实验组较对照组，集落形成速度慢，数目少，克隆形成能力 弱。结论 : MDA-MB-231 细胞 系 WNT6 基 因 表 达 沉 默 ， 可 抑 制 乳 腺 癌MDA-MB-231 细胞系的生长与增殖。关键词:WNT6；siRNA；乳腺癌；Wnt/β-Catenin 信号传导通路2The effect of WNT6 down regulation on the proliferation and growth of MDA-MB-231 cellsAbstractsObjectives:To investigate the effect of WNT6 down regulation on proliferation and growth of breast cancer cells (MDA-MB-231),and define the function of WNT6 in Triple Negative Breast Cancer(TNBC),for illuminating the molecular mechanisms of TNBC.Methods:We chose triple negative breast cancer MDA-MB-231 cell line as the experimental subject,the WNT6 as the target. In our essay, we designed four groupsincluding siRNAl、siRNA2、siRNA3, which with different targeted sequences, and thescramble siRNA(scRNA). We transfected the siRNA into MDA - MB - 231 cells through Lipofectamin 2000 respectively. Then detected the expression of WNT6 by RT-PCR and Western bloting , at last chose the group which one has the best silence effect as experimental group. Using RT-PCR and Western bloting assay to detect the expression of WNT6 of experimental group on the mRNA and protein level, and the not transfection cells and scRNA as control. MTT assay was used to estimate the effect of WNT6 down regulation on proliferation and growth of MDA-MB-231 cells, thereby drew the growth curve in each group. Finally, soft AGAR colony experiment was used to observe anchor dependenciest of cell lines for each group.Results:We discussed the expression of mRNA for WNT6 in each group analysed by RT-PCR .It was observed that WNT6 could be obviously down-regulated in the groupswhich are transfected siRNAl 、 siRNA2 、 siRNA3. Compared to the cell lineMDA-MB-231 witho