NK细胞培养及杀伤前列腺癌细胞效率的研究

Study on NK cell culture and killing efficiency of prostate cancer cells

目的 建立一种高效的NK细胞体外扩增培养方法;通过检测NK细胞杀伤3种前列腺癌(PCa)细胞株的效率,鉴定其用于后期治疗PCa的可行性,为应用NK细胞过继回输治疗PCa的研究奠定基础。方法 通过细胞因子对外周血单个核细胞(PBMCs)的诱导扩增,培养出NK细胞并用流式细胞术检测NK细胞所占比例以及扩增倍数;通过CCK8法检测NK细胞杀伤3种PCa细胞株PC3、LNCaP、DU145的效率及效靶比。结果 NK细胞诱导扩增成功,PBMCs诱导扩增后,NK细胞由最初的(10.97±3.28)%,扩增到占比(83.20±8.54)%以上,细胞总数扩增倍数为(251.66±19.05)倍,NK细胞扩增倍数为(1 940.17±402.22)倍。NK细胞对3种靶细胞的杀伤率差异极显著(P<0.01);NK细胞对LNCaP的杀伤效率大于对DU145的杀伤效率;对于PC3没有杀伤作用,反而促进其生长。NK细胞对PCa细胞株的最高杀伤效率分别为LNCaP(92.03±9.95)%、DU145(72.40±8.30)%、PC3(26.69±10.56)%。结论 采用细胞因子诱导扩增PBMCs的方法能够获得纯度较高、数量足够的NK细胞,为NK细胞的临床应用奠定基础;NK细胞对3种PCa细胞系的杀伤率差异极显著,为后续进行深入研究探索原因及作用机制提供了方向。

Objective To establish an efficient method of NK cell expansion and culture in vitro.The effectiveness of NK cells in killing three prostate cancer(PCa)cell lines was tested to evaluate its feasibility in the later treatment of PCa,which laid a foundation for the application of adoptive NK cell transfusion for PCa.Methods The NK cells were cultured by inducing and amplifying PBMCs with cytokines,and the proportion and amplification of NK cells were detected by flow cytometry.The efficiency and effect-target ratio of NK cells against three prostate cancer cell lines PC3,LNCaP and DU145 were measured by CCK8 method.Results The NK cells were successfully amplified.After PBMCs induction,NK cells were amplified from the initial 10.97%±3.28%to more than 83.20%±8.54%,and the total number of cells was(251.66±19.05)times.The amplification factor of NK cells was(1940.17±402.22).The killing rate of NK cells to the three kinds of target cells was significantly different(P<0.01).The killing efficiency of NK cells against LNCaP was greater than that against DU145.It has no killing effect on PC3,but promotes its growth.The highest killing efficiency of NK cells against prostate cancer cells were LNCaP 92.03%±9.95%,DU14572.40%±8.30%and PC326.69%±10.56%,respectively.Conclusion The proliferation of PBMCs induced by cytokines can obtain high purity and sufficient number of NK cells,which lays a foundation for the clinical application of NK cells.The killing rate of NK cells to the three prostate cancer cells was significantly different,which provided a direction for further research to explore the causes and mechanism of action.