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目的:观察丙戊酸钠(valproic acid,VPA)对人肾癌细胞主要组织相容性复合体Ⅰ类相关链A/B(major histocompatibility complex classⅠ-related chainA/B,MICA/B)表达的影响,比较肾癌细胞经VPA处理前后自然杀伤(natural killer,NK)细胞对其杀伤作用的差异。方法:用0.5～8.0mmol/LVPA分别处理人肾癌786-O和ACHN细胞后,应用FCM检测不同浓度VPA对细胞活力的影响。选择4.0mmol/LVPA处理肾癌细胞,通过实时荧光定量PCR和FCM分别检测肾癌细胞MICA/BmRNA和蛋白的表达水平;钙黄绿素释放法和FCM分别检测VPA处理肾癌细胞后NK细胞对其的杀伤作用以及NK细胞的脱颗粒行为。结果:8.0mmol/LVPA处理肾癌细胞48h后,对肾癌细胞活力的影响大于其他处理组(P<0.05);4.0mmol/LVPA处理组肾癌细胞MICA/BmRNA和蛋白的表达水平均明显高于未经VPA处理的对照组(P<0.05);4.0mmol/LVPA处理组NK细胞对肾癌细胞的杀伤率明显高于未经VPA处理的对照组(P<0.05),且杀伤率的升高可被抗NKG2D(natural-killer group2,memberD)抗体特异性拮抗;肾癌细胞经4.0mmol/LVPA处理后,其激活NK细胞的脱颗粒作用明显强于未经VPA处理的对照组(P<0.05)。结论:VPA可以明显上调肾癌细胞MICA/B的表达,增强NK细胞对其的杀伤活性。
Objective: To observe the effect of valproic acid (VPA) on the expression of major histocompatibility complex class Ⅰ-related chain A / B (MICA / B) in human renal cell carcinoma cell line The differences of killing effect of natural killer (NK) cells on renal cell carcinoma cells before and after VPA treatment were compared. Methods: Human renal carcinoma 786-O and ACHN cells were treated with 0.5-8.0 mmol / L LVPA respectively. The effect of different concentrations VPA on cell viability was detected by FCM. The expression of MICA / B mRNA and protein in renal cancer cells was detected by real-time fluorescence quantitative PCR and FCM respectively after treatment with 4.0 mmol / L LVPA. Calcein release assay and FCM were used to detect the expression of MICA / Killing effect and degranulation behavior of NK cells. Results: After treated with 8.0mmol / LVPA for 48h, the effect on renal cancer cells was greater than that in other treatment groups (P <0.05). The expression levels of MICA / B mRNA and protein in renal cancer cells treated with 4.0mmol / LVPA were significantly higher VPA-treated control group (P <0.05). The killing rate of NK cells in 4.0 mmol / LVPA group was significantly higher than that of the control group without VPA treatment (P <0.05), and the killing rate High can be antagonized by anti-NKG2D (member-D) antibody. The degranulation of NK cells activated by 4.0 mmol / LVPA of renal cancer cells was significantly stronger than that of the control group without VPA treatment (P < 0.05). Conclusion: VPA can upregulate the expression of MICA / B in renal carcinoma cells and enhance the killing activity of NK cells.