尼妥珠单抗联合顺铂对三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用及其机制研究

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[目的]探讨尼妥珠单抗(泰欣生)联合顺铂对三阴性乳腺癌细胞的增殖抑制作用及可能机制。[方法]100μg/ml泰欣生、5μg/ml顺铂单独和联合作用三阴性乳腺癌MDA-MB-231细胞24h、48h后,采用MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Western Blot技术检测ERK1/2、pERK1/2及BRCA1蛋白表达水平。[结果]泰欣生和顺铂联合作用MDA-MB-231细胞48h后,细胞增殖抑制达82.53%±6.46%,与其余各组相比,差异有显著性(P<0.01)。顺铂和泰欣生单药作用24h,未能明显诱导MDA-MB-231细胞凋亡;但作用48h后顺铂诱导的凋亡率为14.56%,泰欣生组的细胞凋亡仍不明显。泰欣生联合顺铂作用24h未能明显诱导MDA-MB-231细胞凋亡;48h后,细胞凋亡率明显增高,达37.57%,与其余各组相比差异有显著性(P<0.01)。ERK1/2在对照组和各处理组中的表达无变化;pERK1/2在对照组和顺铂组的表达无变化,在泰欣生组和泰欣生+顺铂组的表达明显降低。顺铂和泰欣生单药处理MDA-MB-231细胞后BRCA1蛋白表达量增加,泰欣生与顺铂联合作用后,BRCA1蛋白表达明显降低。[结论]尼妥珠单抗(泰欣生)与顺铂可协同抑制三阴性乳腺癌细胞,其机制可能是降低BRCA1蛋白的表达。 [Objective] To investigate the inhibitory effect of nimotuzumab (Taixin Health) combined with cisplatin on the proliferation of triple-negative breast cancer cells and its possible mechanism. [Methods] MDA-MB-231 cells were treated with 100μg / ml Taixinsheng and 5μg / ml cisplatin for 24 hours and 48 hours respectively. The cell proliferation was detected by MTT assay. The apoptosis was detected by flow cytometry. Western Blot detection of ERK1 / 2, pERK1 / 2 and BRCA1 protein expression levels. [Result] The inhibitory effect of Tacsin and cisplatin on the proliferation of MDA-MB-231 cells was 82.53% ± 6.46% at 48h, which was significantly different from other groups (P <0.01). The effect of cisplatin and tacrine alone for 24 h did not induce the apoptosis of MDA-MB-231 cells obviously. However, the apoptotic rate induced by cisplatin was 14.56% . The apoptosis of MDA-MB-231 cells was significantly inhibited by TAXT combined with cisplatin for 24h, and the apoptosis rate was significantly increased to 37.57% after 48h, which was significantly different from other groups (P <0.01) . The expression of ERK1 / 2 in control group and each treatment group had no change. The expression of pERK1 / 2 in control group and cisplatin group had no change, and the expression of ERK1 / 2 was significantly lower in Taixinsheng group and Taixinsheng + cisplatin group. The expression of BRCA1 protein in MDA-MB-231 cells treated with cisplatin and tamsulosin monotherapy increased significantly, and the expression of BRCA1 protein decreased significantly after combined with cisplatin. [Conclusion] Nepalezizumab and cisplatin can synergistically inhibit triple negative breast cancer cells, and its mechanism may be to decrease the expression of BRCA1 protein.
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