Huang Yu-Xian, Wang Yang, Sun Ming, Zhou Xue-Yun, Deng Lan, Guo Kun-Yuan
Department of Hematology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Jul;29(7):1329-32.
To investigate the mechanism underlying the effects of matrine in enhancing the cytotoxic sensitivity of CNE2/DDP cells highly expressing ATP-binding cassette superfamily G member 2 (ABCG(2)(High)) to allogenic natural killer (Allo-NK) cells.
ABCG(2)(High) CNE2/DDP cells and Allo-NK cells were isolated by magnetic activated cell sorting (MACS). Flow cytometry was used to evaluate the purity of the isolated cells and the expression of NKG2D ligands on the target cells before and after incubation with matrine. The cytotoxic sensitivity of the treated and non-treated ABCG(2)(High) CNE2/DDP cells to Allo-NK cells was measured by LDH releasing assay.
The expression rate of ABCG2 was (91.40-/+2.32)% in ABCG(2)(High) CNE2/DDP cells. More than 90% of the isolated NK cells were identified to be CD3(-)CD16(+)CD56(+) cells. The expression rates of MICA, MICB, ULBP1, ULBP2, and ULBP3 on the target cells incubated with matrine increased from (2.92-/+0.33)%, (4.27-/+0.33)%, (5.80-/+0.62)%, (11.10-/+3.15)%, and (7.75-/+1.14)% to (11.30-/+0.89)%, (14.29-/+2.61)%, (12.56-/+1.06)%, (43.24-/+4.43)%, and (12.77-/+1.06)%, respectively. At the E: T ratio of 10:1 and 20:1, the cytotoxic sensitivity of ABCG(2)(High) cells to Allo-NK cells increased from (15.32-/+1.34)% and (27.26-/+6.81)% in un-treated cells to (28.53-/+1.37)% and (42.72-/+2.80)% in matrine-treated cells, respectively, showing significant differences in the cytotoxic sensitivity of the target cells in both groups produced by matrine treatment (F=29.05, P=0.000).
Matrine can up-regulate the expressions of NKG2D ligands (MICA/B and ULBP1-3) in ABCG(2)(High) nasopharyngeal carcinoma cells, which results in increased cytotoxic sensitivity of the cells to Allo-NK cells.
探讨苦参碱增强高表达ATP结合盒超家族G成员2（ABCG(2)(High)）的CNE2/DDP细胞对同种异体自然杀伤（Allo-NK）细胞细胞毒性敏感性的作用机制。
通过磁珠分选法分离ABCG(2)(High) CNE2/DDP细胞和Allo-NK细胞。采用流式细胞术评估分离细胞的纯度以及苦参碱孵育前后靶细胞上NKG2D配体的表达。通过乳酸脱氢酶释放试验检测处理和未处理的ABCG(2)(High) CNE2/DDP细胞对Allo-NK细胞的细胞毒性敏感性。
ABCG(2)(High) CNE2/DDP细胞中ABCG2的表达率为(91.40±2.32)%。超过90%的分离NK细胞被鉴定为CD3(-)CD16(+)CD56(+)细胞。与苦参碱孵育后的靶细胞上MICA、MICB、ULBP1、ULBP2和ULBP3的表达率分别从(2.92±0.33)%、(4.27±0.33)%、(5.80±0.62)%、(11.10±3.15)%和(7.75±1.14)%增加到(11.30±0.89)%、(14.29±2.61)%、(12.56±1.06)%、(43.24±4.43)%和(12.77±1.06)%。在效靶比为10:1和20:1时，ABCG(2)(High)细胞对Allo-NK细胞的细胞毒性敏感性分别从未处理细胞的(15.32±1.34)%和(27.26±6.81)%增加到苦参碱处理细胞的(28.53±1.37)%和(42.72±2.80)%，显示苦参碱处理使两组靶细胞的细胞毒性敏感性存在显著差异（F=29.05，P=0.000）。
苦参碱可上调ABCG(2)(High)鼻咽癌细胞中NKG2D配体（MICA/B和ULBP1-3）的表达，从而导致细胞对Allo-NK细胞的细胞毒性敏感性增加。
