EphrinA1-caspase-3对乳腺癌细胞的体外靶向性抑制作用

Inhibitory effect of EphrinA1-caspase-3 on breast cancer cells in vitro

目的:研究重组EphrinA1-caspase-3对体外乳腺癌细胞是否有靶向性抑制作用。方法:将重组EphrinA1-caspase-3感染人乳腺细胞,MTT法检测重组Ad-EphrinA1-caspase-3感染上清对EphA2阳性乳腺癌细胞的体外增殖的影响,采用流式细胞仪检测Ad-EphrinA1-caspase-3感染上清对乳腺癌细胞凋亡的影响,采用抗体中和实验检测感染上清中EphrinA1-caspase-3对靶细胞杀伤效应的特异性。结果:乳腺癌细胞呈贴壁生长,细胞中可见高水平的EphA2受体及EphrinA1表达。Ad-EphrinA1-caspase-3感染上清对乳腺癌细胞增殖表现出强烈的抑制作用,生长抑制率达48.9%。用流式细胞仪检测培养72h后稀释倍比为1∶1、1∶2、1∶4、1∶8、1∶16上清对应的凋亡率分别为36%、24%、18%、8%、7%,显著高于对照组凋亡率4%,且中和抗体能有效的减弱重组EphrinA1-caspase-3对EphA2阳性乳腺癌细胞的杀伤作用。结论:感染EphrinA1-caspase-3的T淋巴细胞能够对体外乳腺癌细胞起到靶向性抑制作用。

To study whether the recombinant EphrinA1-caspase-3 has a targeted inhibitory effect on breast cancer cells in vitro.Methods:The breast cancer cells were isolated,cultured and identified.Then the recombinant EphrinA1-caspase-3 was used to infect human breast cells,and the effect of recombinant Ad-EphrinA1-caspase-3 on the proliferation of EphA2 positive breast cancer cells in vitro were tested by MTT.The effect of Ad-EphrinA1-caspase-3 supernatant on the apoptosis of breast cancer cells was detected by flow cytometry.Finally,the specific killing effect of EphrinA1-caspase-3 on the target cells was examined by antibody neutralization experiments.Results:Breast cancer cells showed adherent growth,and overexpression of EphA2 receptor and EphrinA1 were observed in breast cancer cells.The Ad-EphrinA1-caspase-3 infection supernatant showed a strong inhibitory effect on the proliferation of breast cancer cells,and the growth inhibition rate was 48.9%.72 hours after culture,the apoptosis rates corresponding to the dilution ratios of 1∶1,1∶2,1∶4,1∶8 and 1∶16 supernatants were 36%,24%,18%,8%,and 7%,respectively,which were significantly higher than those of the control group.The mortality rate was 4%(P<0.05).And neutralizing antibody could effectively reduce the killing effect of recombinant EphrinA1-caspase-3 on EphA2 positive breast cancer cells.Conclusion:The T lymphocytes infected with EphrinA1-caspase-3 has a targeted inhibitory effect on breast cancer cells in vitro.