沉默UHRF1对乳腺癌细胞增殖和转移的影响

目的 探讨UHRF1对乳腺癌MDA-MB-231细胞增殖以及侵袭的影响及其相关机制。方法 采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测沉默UHRF1基因后对乳腺癌MDA-MB-231细胞活力的影响;应用克隆形成实验检测沉默UHRF1后对乳腺癌MDA-MB-231细胞存活的影响;吖啶橙-溴乙锭(AO/EB)检测沉默UHRF1后对乳腺癌MDA-MB-231细胞凋亡的影响;Caspase-3活性试剂盒检测沉默UHRF1后乳腺癌细胞Casapse-3活性的变化;Western blot法检测细胞中凋亡相关蛋白Bcl-2、Bax、Bad、p-Bad、XIAP、p53、p21^Cip1/Waf1和p16^INK4a的表达;应用Transwell实验研究沉默UHRF1对MDA-MB-231细胞侵袭能力的影响;Wound Healing实验研究沉默UHRF1后对其迁移能力的影响。结果 沉默UHRF1使乳腺癌MDA-MB-231细胞活力降低;克隆形成实验结果显示沉默UHRF1后MDA-MB-231细胞存活能力降低,AO/EB染色显示沉默UHRF1促进MDA-MB-231细胞凋亡。同时Caspase-3活性实验结果显示沉默UHRF1后乳腺癌MDA-MB-231细胞Caspase-3的活性增加;Western blot结果显示,沉默UHRF1后,能够使凋亡蛋白Bad、XIAP和Bax的表达上调,同时抗凋亡蛋白p-Bad,Bcl-2的表达下调,也使p53,p21Cip1/Waf1,p16^INK4a蛋白表达升高;Transwell以及Wound Healing实验证明沉默UHRF1能够抑制乳腺癌MDA-MB-231细胞侵袭和迁移。结论 沉默UHRF1能够抑制乳腺癌MDA-MB-231细胞活力和存活,并抑制乳腺癌MDA-MB-231的侵袭和迁移。沉默UHRF1通过调控p53,p21Cip1/Waf1,p16^INK4a信号发挥作用。

Effect of silencing UHRF1 on proliferation and metastasis of breast cancer cells

Objective The of this study was to investigate the effect of UHRF1 on the proliferation and metastasis of breast cancer MDA-MB-231 cells and its mechanism.Methods The effect of silencing UHRF1 gene on the viability of MDA-MB-231 cells was detected by MTT assay.Colony formation assay was performed to analyze the effect of silencing UHRF1 on cell survival of MDA-MB-231 cells.The effect of silencing UHRF1 on the apoptosis of MDA-MB-231 cells was detected by acridine orange-ethidium bromide (AO / EB).Caspase-3 activity kit was used to detect the expression of caspase-3 in MDA-MB-231 cells.The expressions of Bcl-2,Bax,Bad,p-Bad,XIAP,p53,p21^Cip1/Waf and p16^INK4a were detectedby Western blot.The abilities of invasion and migration of MDA-MB-231 cells silenced by UHRF1were examined by Transwell and Wound healing assays,respectively.Results Silencing UHRF1 significantlydecreased the viability of MDA-MB-231 cells.Silencing UHRF1 decreased colony formation in MDA-MB-231 cells.Depletion of UHRF1 resulted in apoptosis inducedin MDA-MB-231 cells,showing nuclear morphological changes by AO/EB staining and increasing caspase-3 activity.After knockdown of UHRF1,the expression of Bad,XIAP,Bax,p53,p21^Cip1/Waf1 and p16^INK4a was up-regulatedand down-regulated the expression of p-Bad and Bcl-2 in MDA-MB-231 cells.Transwell and wound healing assays demonstrated that silencing UHRF1 could decrease metastasisin MDA-MB-231 cells.Conclusion Silencing UHRF1 can inhibit the viability and survival of MDA-MB-231 cells,and inhibit the invasion and migration of MDA-MB-231 cells regulated by p53/p21^Cip1/Waf1/p16^INK4asignalings.