PI3K抑制剂BKM120体外抗结外鼻型NK/T细胞淋巴瘤的作用

Effect of PI3K Inhibitor BKM120 on Extranodal NK / T Cell Lymphoma In Vitro

【目的】探讨PI3K抑制剂BKM120体外对结外鼻型NK/TL细胞淋巴瘤(ENKTL)的疗效。【方法】BKM120处理ENKTL细胞SNK6和SNT8后,CCK8试剂盒检测细胞增殖,PI单染色法检测细胞周期,Annexin V/PI双染色法检测细胞凋亡,Western Blotting检测p-PI3K、AKT、p-AKT以及细胞周期相关蛋白的表达情况。BKM120分别与依维莫司、阿霉素和吉西他滨联合处理SNK6和SNT8细胞,金正均"Q"值法分析联合作用效应。【结果】BKM120能够有效抑制SNK6、SNT8细胞的增殖,其IC50值分别为(0.23±0.013)μmol/L和(0.17±0.011)μmol/L。BKM120能使SNK6、SNT8细胞阻滞于G1期,1.0μmol/L的BKM120使SNK6、SNT8细胞G1期比例较对照组分别增加(12.53±1.31)%(P=0.013)和(30.15±2.79)%(P=0.001)。0.5μmol/L和1.0μmol/L的BKM120处理后,SNK6和SNT8细胞均无明显凋亡。此外,BKM120能够降低SNK6、SNT8细胞的p-PI3K、AKT、p-AKT和Cyclin D1的表达。BKM120分别与依维莫司、阿霉素和吉西他滨联合处理SNK6和SNT8细胞,Q值处于0.85至1.15之间,BKM120与mTOR抑制剂、化疗药物联合应用时具有相加作用。【结论】BKM120能够有效抑制ENKTL细胞SNK6和SNT8的增殖,该抑制效应的机制之一是将细胞周期阻滞于G1期。BKM120分别与依维莫司、阿霉素和吉西他滨联合应用,具有相加作用。

[Objective] To investigate the effect of PI3K inhibitor BKM120 on extranodal NK/T cell lymphoma （ENKTL） in vitro.[Methods] ENKTL cell lines SNK6 and SNT8 were treated with BKM 120,subsequently the cellular proliferation was tested by Cell Counting-kit 8.The cell cycle and apoptosis analyses were tested by FACS with either PI single dying or Annexin V/PI double dying.Western Blotting was performted to detect the expression of p-PI3K,AKT,p-AKT,and cell cycle related proteins.The effects of BKM120 combined with everolimus,adriamycin,and gemcitabine separately on SNK6 and SNT8 were also detected by using the Zheng-Jun Jin ＂Q＂ method.[Results] BKM120 could inhibit the proliferation of SNK6 and SNT8 cells effectively.The IC50 of BKM120 on SNK6 and SNT8 were 0.23 ± 0.013 μmol/L and 0.17 ± 0.011 μmol/L,respectively.BKM120 could induce cell cycle arrest in SNK6 and SNT8 cells.After treated with 1.0 μmol/L BKM120,the percentage of G1 phase increased （12.53 ± 1.31）% （P =0.013） in SNK6 and （30.15±2.79）% （P=0.001） in SNT8.No significant apoptosis was detected in SNK6 and SNT8 cells after treated with BKM120 at the concentrations of 0.5 μmol/L and 1.0 μmol/L.The expression level of p-PI3K,AKT,p-AKT,and Cyclin D1 in SNK6 and SNT8 were decreased by BKM120.The Q value was 0.85 to 1.15,when BKM120 was combined with everolimus,adriamycin,and gemcitabine on SNK6 and SNT8,respectively.BKM120 has additive effect with mTOR inhibitor and chemotherapy agents.[Conclusions] BKM120 can inhibit the proliferation of SNK6 and SNT8 cells effectively,and one mechanism may be to arrest the cell cycle into G1 phase.There are additive effects when BKM120 are combined with everolimus,adriamycin,and gemcitabine.