摘要
目的研究生长抑制因子5(ING5)基因对人乳腺癌细胞Bcap-37增殖、凋亡、迁移和细胞周期的影响。方法稳定转染p EGFP-N1-ING5真核表达载体和p EGFP-N1空载体至Bcap-37细胞,荧光显微镜观察绿色荧光蛋白的表达,RT-PCR方法筛选ING5高表达细胞株。将Bcap-37-ING5细胞作为实验组,Bcap-37-GFP细胞为空载体组,Bcap-37为空白对照组。采用MTT法检测ING5对细胞增殖的影响,流式细胞仪检测细胞周期及凋亡情况;细胞划痕实验和Transwell实验检测细胞迁移情况。结果稳定转染后获得稳定表达GFP标记的ING5蛋白的Bcap-37细胞株。ING5过表达可以抑制乳腺癌Bcap-37细胞增殖并导致G2期阻滞,诱导细胞凋亡,抑制细胞迁移(P<0.05)。结论 ING5过表达可逆转乳腺癌Bcap-37细胞的恶性表型,可作为乳腺癌预后判断和基因治疗的分子靶标。
Objective To investigate the effects of inhibitor of growth 5（ING5） gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells. Methods The eukaryotic ING5-expressing plasmid and GFPempty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment. Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability（P〈0.05）. Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2017年第1期12-16,共5页
Medical Journal of Chinese People's Liberation Army
基金
辽宁省科学技术计划项目(2015020339)~~
