GINS2蛋白对三阴性乳腺癌细胞的作用及机制分析

Effects and mechanism of GINS2 on triple-negative breast cancer cells and tumor stem cells

目的分析GINS2蛋白对三阴性乳腺癌细胞及肿瘤干细胞的作用,并初步探索其作用机制。方法选取三阴性乳腺癌(triple negative breast cancer,TNBC)细胞系MDA-MB-436和MDA-MB-157细胞,构建正常对照组及GINS2敲低组。通过集落形成和细胞增殖测定比较细胞生长能力,细胞侵袭试验比较侵袭能力,Aldhhi细胞分选及成球分析比较两组细胞"干性"的差异,流式细胞仪分析细胞周期变化。通过蛋白质印迹法和免疫共沉淀实验研究GINS2在乳腺癌细胞中相互作用的蛋白。结果 TNBC细胞中转染siRNA后,GINS2降低导致TNBC细胞的生长能力、侵袭能力、肿瘤干细胞"干性"均明显下降,GINS2下降导致细胞在G2期持续积累,在G1期数量减少。另外,GINS2与JUN蛋白在细胞内相互结合。结论敲低GINS2可抑制TNBC生长和转移,是未来的特异性治疗靶点。

Objective To analyze the effects of GINS2 on triple-negative breast cancer cells and tumor stem cell and explore the mechanism of these effects.Methods The triple negative breast cancer(TNBC)cell lines including MDA-MB-436 and MDAMB-157 were selected,and the control group and GINS2 knockdown group cells were constructed.The differences between the two groups were analyzed and compared.Colony formation and cell proliferation assay were used to compare cell growth ability,cell invasion assay to compare invasive ability,Aldhhi cell sorting and sphere assay to compare the stemness of the two groups of cancer stem cells,and cell cycle changes were analyzed by ow cytometry.The cellular proteins in two groups were extracted,and proteins interacting with GINS2 in breast cancer cells were studied by Western blotting and co-immunoprecipitation experiment.Results After transfection of siRNA in TNBC cells,the decrease of GINS2 resulted in the decrease of growth ability,invasive ability,and stemness of tumor stem cells.The decrease of GINS2 caused the cells to accumulate in G2 phase and the number of cells in G1 phase decreased.At the same time,the study found that GINS2 and JUN proteins had specic interactions in the cells.Conclusion Knockdown of GINS2 inhibits the growth and metastasis of TNBC,which could be a therapeutic target.