紫杉醇顺铂对HCC1937人乳腺癌细胞MAPK信号通路影响

Role of the MAPK Signal Pathway in Paclitaxel- and Cisplatin-induced Apoptosis in HCC1937 Breast Cancer Cells

目的:研究紫杉醇、顺铂对BRCA1基因缺陷型三阴性乳腺癌细胞HCC1937的增殖抑制作用及与MAPK信号通路的关系。方法:采用CCK-8试剂盒检测紫杉醇、顺铂分别对HCC1937细胞、MCF-7细胞的50%抑制浓度(IC_(50));采用流式细胞仪和Western blot分别检测两药作用HCC1937细胞48 h后的细胞周期和MAPK通路蛋白表达状况。结果:HCC1937细胞(IC_(50) 6～9μg/mL)对顺铂敏感性显著高于MCF-7细胞(IC_(50) 18～20μg/mL)(P<0.01);而HCC1937细胞(IC_(50) 3～4.6μg/mL)对紫杉醇的敏感性则明显低于MCF-7细胞(IC_(50) 0.12～0.3μg/mL)(P<0.01)。紫杉醇使HCC1937细胞阻断在G_2～M期,呈剂量-效应趋势,顺铂使其阻断在G_0/G_1期;紫杉醇、顺铂作用HCC1937细胞48 h后,P-JNK和P-P38蛋白表达显著增加,顺铂组P-ERK蛋白表达较对照组明显降低。结论:1)三阴性乳腺癌细胞HCC1937对顺铂的敏感性明显优于紫杉醇;2)紫杉醇、顺铂皆可激活HCC1937细胞JNK/SAPK、P38通路,同时不同浓度的顺铂可以抑制ERK通路激活。

Objective： This study aimed to investigate the inhibitory effects of paclitaxel and cisplatin against the triple-negative breast cancer cell line HCC 1937 in vitro, and to explore the underlying mechanisms in relation to the MAPK signaling pathway. Meth- ods： A Cell Counting Kit-8 assay was used to detect the 50% inhibitory concentration （ICs0） of paclitaxel and cisplatin on HCC1937 and MCF-7 cells. Flow cytometry and Westem blot analysis were used to determine the cell cycle distribution and MAPK signaling path- way protein expression after incubating the HCC 1937cells with paclitaxel or cisplatin for 48 h. Results： The HCC 1937 cells were sig- nificantly more sensitive to cisplatin （ICs0 = 6-9 pg/ml） than MCF-7 cells （ICs0 = 18-20 μg/ml） （P 〈 0.01）, whereas the HCC1937 cells were less sensitive to paclitaxel （ICs0 = 3-4.6 μg/ml） than the MCF-7 cells （ICs0 = 0.12-0.3 μg/ml） （P 〈 0.01）. Paclitaxel caused the MCF-7 cells to arrest at the GJM phase in a concentration-dependent manner, whereas cisplatin acted by arresting cells at the G0/G, phase. P-JNK and P-P38 protein expression significantly increased after the cells were treated with the two drugs for 48 h compared with the control group. However, P-ERK protein expression in the cisplatin group was significantly lower than in the control group for the HCC1937cells. Conclusion： HCC1937 cells have higher chemosensitivity to cisplatin than to paclitaxel. Paclitaxel and cisplatin can both activate the JNK/SAPK and P38 signaling pathways in HCC1937 cells, whereas different cisplatin concentrations can inhibit the activation of the ERK signaling pathway.