树突状细胞与自然杀伤细胞共同作用对人黑色素瘤A375细胞端粒酶反转录酶mRNA的间接影响

Indirect effect of dendritic cells and natural killer cells on the human telomerase reverse transcriptase mRNA in human melanoma cell line A375

目的 观察树突状细胞（DC）与自然杀伤（NK）细胞体外共同作用对人黑色素瘤A375细胞人端粒酶反转录酶（hTERT） mRNA的影响.方法 从健康人外周血中获得DC、NK细胞,在transwell培养体系中用DC、NK细胞单独或共同作用的微环境作用A375细胞,采用反转录PCR检测A375细胞hTERT mRNA表达的变化.结果 transwell下室的DC与NK细胞单独或共同作用均能间接抑制上室A375细胞hTERT mRNA的表达,DC联合NK细胞的抑制作用更强,且对A375细胞hTERTmRNA的抑制具有时间依赖性.结论 DC、NK细胞能间接抑制人恶性黑色素瘤A375细胞hTERTmRNA的表达,DC和NK共同作用有更强的抑制作用.

Objective To investigate the effect of dendritic cells （DC） and natural killer （NK） cells on the human telomerase reverse transcriptase （hTERT） mRNA in human melanoma cell line A375 in vitro.Methods A co-culture system was established by transwell culture system,in which DC and NK cells obtained from peripheral blood of healthy donors were added to lower chamber for 24 h separately or together.Then A375 cells was co-cultured in upper chamber of the transwell for 24 h,48 h and 72 h.A375 cells were collected and hTERT mRNA were analyzed by reverse transcription-polymerase chain reaction （RT-PCR） assay.Results DC and NK cells reduced the expression of hTERT mRNA in A375 cells separately or together （DC/NK）.However,the reduction of hTERT mRNA expression in DC/NK group was more apparently than that in DC or NK group.Furthermore,DC/NK group reduced the hTERT mRNA expression in a timedependent manner.Conclusion This is the first time to demonstrate that DC and NK cells have an indirect influence on the hTERT mRNA expression in human melanoma cell line A375,and there is an synergetic effect between DC and NK.The immune microenvironment created by interaction between DC and NK has a stronger inhibitory effect.