Markers associated to NK cytolytic activity are in a great need to regulate NK cell immunotherapy products. We assume that biomarkers which response to cytolysis will change their transcription, expression or secretion. To find NK-92 indicator to cytolytic activity, we have evaluated the potential markers by quantifying the expression of well-known cytotoxicity functional molecules (cytokine IFN-γ, Granzyme B, perforin, CD69 and CD107a), and explored candidate markers by a sweeping transcription picture of NK-92 using a direct cytolysis model (incubation with K562). We found that IFN-γ secretion was highly correlated to cytotoxicity of NK-92, neither Granzyme B, perforin secretion, nor CD69, CD107a positive population were upregulated by K562 stimulation. RNAseq revealed 432 genes expression changed during cytolysis, several genes (BIRC3, CSF2, VCAM1 and TNFRSF9) mRNA expression were validated by real time RT-PCR under K562 being killed or protected from being killed conditions. Results suggested IFN-γ secretion, BIRC3 and TNFRSF9 transcription in NK-92 were responsive to K562 cytolysis. In a word, our results confirmed one marker and reveal an array of novel candidate markers associated with NK-92 cytotoxicity. Further studies are greatly needed to determine the roles these new makers play in NK-92 cytolysis process.

非常需要与 NK 细胞溶解活性相关的标志物来调节 NK 细胞免疫治疗产品。我们假设对细胞溶解有反应的生物标志物会改变它们的转录、表达或分泌。为了找到 NK-92 细胞溶解活性的指标，我们通过量化众所周知的细胞毒性功能分子（细胞因子 IFN-γ、颗粒酶 B、穿孔素、CD69 和 CD107a）的表达来评估潜在标志物，并通过扫描探索候选标志物使用直接细胞溶解模型（与 K562 孵育）的 NK-92 转录图片。我们发现 IFN-γ 分泌与 NK-92 的细胞毒性高度相关，颗粒酶 B、穿孔素分泌和 CD69、CD107a 阳性群体均不会被 K562 刺激上调。RNAseq 显示 432 个基因表达在细胞溶解过程中发生了变化，几个基因（BIRC3、CSF2、VCAM1 和 TNFRSF9) mRNA 表达通过实时 RT-PCR 在 K562 被杀死或被保护免于被杀死的条件下进行验证。结果表明 NK-92 中的 IFN-γ 分泌、BIRC3 和 TNFRSF9 转录对 K562 细胞溶解有反应。总之，我们的结果证实了一个标记，并揭示了一系列与 NK-92 细胞毒性相关的新型候选标记。非常需要进一步的研究来确定这些新制造商在 NK-92 细胞溶解过程中的作用。