二烯丙基三硫化物对人神经母细胞瘤细胞SK—N—SH生长的抑制作用

Experimental study on inhibition the growth of neuroblastoma by diallyl trisulfide in vitro

目的探讨二烯丙基三硫化物（DATS）对人神经母细胞瘤细胞SK．N．SH生长的影响及相关机制。方法光镜下观察不同浓度DATS对SK—N—SH细胞生长的影响；细胞分为对照组、DATSI组（10p晷／L）和DATSⅡ组（40肛∥L），DATS作用24、48h后，流式细胞仪测定细胞凋亡；作用72h后，荧光定量聚合酶链反应（FQ—PCR）法测定DATS对SK—N—SH细胞血管内皮生长因子（VEGF）mRNA、细胞间黏附分子一1（ICAM．1）mRNA表达的影响；酶联免疫吸附试验（ELISA）法测定细胞培养上清液VEGF、ICAM．1浓度。结果不同浓度的DATS均能抑制SK—N—SH细胞的生长，小剂量DATS能诱导细胞凋亡；对照组、DATSI组和Ⅱ组VEGFmRNA、ICAM一1mRNA表达量分别为5．974-1．31、4．21±1．06、3．35±0．77（P〈0．05）和4．41±O．94、3．22±1．17、2．69±O．73（P〈0．05）；对照组、DATSI组和Ⅱ组SK—N．SH细胞培养上清液中VEGF、ICAM一1分别为（72．14-26．9）、（51．6±17．3）、（40．3±11．8）μg／L（P〈0．05）和（311．6±83．7）、（274．2±71．0）、200．6±42．9）ng／L（P〈0．05）。结论DATS在体外可抑制神经母细胞瘤细胞SK-N—SH的生长，DATS能诱导SK—N-SH细胞凋亡，下调其VEGFmRNA、ICAM一1mRNA的转录和蛋白表达。

Objective To investigate the efficacy and the possible mechanism of human neuroblas- toma cell line SK-N-SH induced by diallyl trisulfide （DATS） in vitro. Methods Flow cytometry was ap- plyed to test the apoptosis rate of human neuroblastoma cell lines SK-N-SH after DATS was used 24 h and 48 h. SK-N-SH cells were randomly divided into 3 groups： the control group, the DATS group I （DATS 10 trg/L） and group lI （ DATS 40 μg/L）, the role of DATS on the growth of SK-N-SH ceils was evalua- ted after 72 h. The expression of vascular endothelial growth factor （VEGF） mRNA and intercellular adhe- sion molecule-1 （ ICAM-1 ） mRNA of SK-N-SH ceils were detected by fluorescent quantitation ploymerase chain reaction （FQ-PCR）. The VEGF and ICAM-1 concentration of supernatants were measured by enzyme linked immunosorbent assay （ELISA）. Results DATS inhibited the growth of SK-N-SH cells in vitro, small dose of DATS induced apoptosis of SK-N-SH cells （P 〈 O. 05 ）. The expression of VEGF mRNA in the control group, the DATS group I and DAIS group II was （5.97 ± 1.31 ）, （4. 21 ± 1.06） and （3.35±0.77） （P〈0.05） , the expression of ICAM-1 mRNA was （4.41 ±0.94） , （3.22 ±1.17） and （2.69 ＋ 0. 73） （P 〈 0. 05 ）. The VEGF concentration of culture supernatants of SK-N-SH cells was （ 72. 1 ± 26. 9）, （5L 6 ± 17. 3） and （40. 3 ± 11.8） μg,/L in the 3 groups （P 〈0. 05）, and the ICAM-1 concen- tration was （311.6 ± 83.7 ）, （ 274. 2 ± 71.0 ） and （ 200. 6 ＋ 42. 9 ） ng/L （ P 〈 0, 05 ）. Conclusion DAIS can inhibit the growth of human neuroblastoma cell lines SK-N-SH in vitro, which may caused by in- ducing apoptosis and down-regulating the expression of VEGF and ICAM-1.