他尼氟酯诱导乳腺癌细胞凋亡的实验研究

Experimental study of apoptosis induced by Talniflumate in breast cancer cells

目的探讨黏蛋白调节剂他尼氟酯(Talniflumate)对乳腺癌细胞的作用及其与化疗药紫杉醇联用后的协同效应。方法体外培养乳腺癌细胞。采用淋巴细胞增殖活性检测(MTS)法检测不同浓度Talniflumate单独和联用紫杉醇对乳腺癌细胞存活率的影响,流式细胞术检测上述用药对乳腺癌细胞凋亡的影响。Western blot实验检测Talniflumate作用细胞前后其作用靶点葡萄糖氨基转移酶3(GCNT3)在乳腺癌细胞中表达的改变。转录组测序明确Talniflumate作用细胞后相关信号通路的改变。结果Talniflumate以浓度依赖性促进乳腺癌细胞MCF7及MDA-MB-231细胞的凋亡,Talniflumate与紫杉醇联用对MCF7细胞有显著的协同杀灭作用,但这种作用在MDA-MB-231细胞中不明显。Western blot实验检测发现GCNT3在MCF7细胞中高表达,而在MDA-MB-231细胞中痕量表达;Talniflumate可以诱导MCF7细胞中GCNT3蛋白的表达下调,与紫杉醇联用后GCNT3下调更显著。转录组测序提示Talniflumate可引起TNF、p53、SNARE等多个信号通路的表达改变。结论Talniflumate单用可诱导乳腺癌细胞凋亡。Talniflumate与紫杉醇联用对GCNT3高表达乳腺癌细胞如MCF7细胞有显著协同杀灭肿瘤细胞效应。Talniflumate作用机制可能与TNF、p53、SNARE等多个信号通路相关。

Objective To explore the effect of the mucin modulator Talniflumate(Tal)on breast cancer cells and its synergistic effect after combined with the chemotherapy drug paclitaxel(PTX).Methods The breast cancer cells were cultured in vitro.Lymphocyte proliferation activity assay(MTS)was used to detect the effects of different concentrations of Talniflumate alone and paclitaxel on the survival rate of breast cancer cells.The effects of the above drugs on the apoptosis of breast cancer cells were detected by flow cytometry.Western blot was used to detect the expression of glucosamine transferase(GCNT3)(the target of Talniflumate)in breast cancer cells before and after the treatment with Talniflumate.Transcriptome sequencing clarified the changes in related signaling pathways after treatment with Talniflumate.Results Talniflumate promoted the apoptosis of breast cancer cells MCF7 and MDA-MB-231 cells in a concentration-dependent manner.The combination of Talniflumate and paclitaxel had a significant synergistic killing effect in MCF7 cells but not MDA-MB-231 cells.Western blot indicated that GCNT3 was highly expressed in MCF7 cells,while almost no expression in MDA-MB-231 cells;Talniflumate could reduce the expression of GCNT3 in MCF7 cells and after combined with paclitaxel,the expression of GCNT3 was downregulated more significantly.Transcriptome sequencing suggested that Talniflumate can regulate the expression of multiple signaling pathways such as TNF,p53,and SNARE.Conclusions Talniflumate could induce apoptosis of breast cancer cell.Talniflumate combined with paclitaxel has a significant synergistic effect in killing tumor cells in breast cancer cells with high GCNT3 expression such as MCF7.The mechanism of Talniflumate induce apoptosis of breast cancer cells may be related to multiple signaling pathways such as TNF,p53,and SNARE.