Department of Dermatology, Second Affiliated Hospital, Sun Yat-Sen University, Guangdong, China.
Clin Exp Dermatol. 2009 Dec;34(8):e537-43. doi: 10.1111/j.1365-2230.2009.03594.x.
Although diallyl trisulphide (DATS) has been found to induce apoptosis in various tumour cells, its cytotoxicity in melanoma cells has not yet been defined and the molecular pathway by which DATS induces apoptosis is not well understood.
To determine growth inhibition of DATS in human melanoma cells (A375 and M14) by inducing apoptosis, and to investigate the mechanism underlying such effects.
Growth inhibition by DATS was estimated by the tetrazolium assay. Apoptosis induction in DATS-treated cells was assessed by staining with 4',6-diamidino-2-phenylindole (DAPI) and double staining with annexin V and propidium iodide. Expression of Bcl-2, Bax, Bcl-xL/Bcl-xS, cytochrome c release, activation of caspase-9 and poly(ADP-ribose) polymerase (PARP) were determined by western blotting. The activity of caspase-3 was measured using a colorimetric assay.
DATS exerted its cytotoxic effect in a time-dependent and dose-dependent manner by inducing apoptosis in A375 and M14 cells. Expression of Bcl-2 and Bcl-xL was downregulated. Release of cytochrome c and activation of the downstream effectors caspase-3, caspase-9 and PARP were detected after DATS sensitization.
DATS inhibits growth of melanoma cells by inducing apoptosis in association with downregulation of Bcl-2 and Bcl-xL and activation of caspases.
尽管二烯丙基三硫（DATS）已被发现可诱导多种肿瘤细胞凋亡，但它在黑素瘤细胞中的细胞毒性尚未确定，且 DATS 诱导凋亡的分子途径也尚未完全了解。
通过诱导凋亡来确定 DATS 对人黑素瘤细胞（A375 和 M14）的生长抑制作用，并探讨其作用机制。
通过噻唑蓝（MTT）比色法评估 DATS 的生长抑制作用。通过用 4'，6-二脒基-2-苯基吲哚（DAPI）染色和用膜联蛋白 V 和碘化丙啶双重染色来评估 DATS 处理细胞中的凋亡诱导。通过 Western 印迹测定 Bcl-2、Bax、Bcl-xL/Bcl-xS、细胞色素 c 释放、caspase-9 和多聚（ADP-核糖）聚合酶（PARP）的激活。使用比色法测定 caspase-3 的活性。
DATS 通过诱导 A375 和 M14 细胞凋亡，以时间和剂量依赖的方式发挥其细胞毒性作用。Bcl-2 和 Bcl-xL 的表达下调。在 DATS 敏化后检测到细胞色素 c 的释放和下游效应物 caspase-3、caspase-9 和 PARP 的激活。
DATS 通过下调 Bcl-2 和 Bcl-xL 并激活胱天蛋白酶来诱导凋亡从而抑制黑素瘤细胞的生长。
