Zhang Yue, Xie Wen-Peng, Zhang Yong-Kui, Chen Yi-Qiang, Wang Dong-Li, Li Gang, Guan Dong-Hui
First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan, People's Republic of China.
Department of Orthopedics, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, People's Republic of China.
Onco Targets Ther. 2018 Jan 9;11:271-277. doi: 10.2147/OTT.S150933. eCollection 2018.
Diallyl trisulfide (DATS) is a natural organic sulfur compound isolated from garlic that has good anticancer activity according to many previous reports. There are many studies pointing out that DATS can downregulate expression of the glucose-regulated protein 78 (GRP78), which is associated with poor prognosis and drug resistance in various types of human cancers. However, it remains unknown whether DATS has the same effect on human osteosarcoma cells. This study attempted to clarify the potential molecular mechanisms of the action of DATS in human osteosarcoma Saos-2 cells.
We used an inverted phase microscope and immunofluorescent staining to observe the morphological changes of Saos-2 cells after being cultured in different concentrations of DATS (0, 25, 50, and 100 μM) for 24 h, or for four time periods (24, 48, 72, and 96 h) in the same DATS concentration (50 μM). Quantitative real-time polymerase chain reaction and Western blot were used to detect the expression level of GRP78 mRNA and proteins in Saos-2 cells. GRP78 expression was suppressed in Saos-2 cells by utilizing small-interfering RNA, and the cells were subsequently used to study the anti-proliferative effects of DATS treatment.
The expression level of GRP78 mRNA and proteins was significantly downregulated due to the increased concentration and effective times of DATS (<0.05). In addition, there were significant associations between GRP78 silencing and cell proliferation (<0.05) of DATS treatment.
These results indicate that DATS inhibits the growth of human osteosarcoma Saos-2 cells by downregulating the expression of GRP78.
二烯丙基三硫醚(DATS)是一种从大蒜中分离出的天然有机硫化合物,根据以往许多报道,其具有良好的抗癌活性。许多研究指出,DATS可下调葡萄糖调节蛋白78(GRP78)的表达,而GRP78与多种人类癌症的不良预后和耐药性相关。然而,DATS对人骨肉瘤细胞是否具有相同作用尚不清楚。本研究试图阐明DATS在人骨肉瘤Saos-2细胞中作用的潜在分子机制。
我们使用倒置相差显微镜和免疫荧光染色观察Saos-2细胞在不同浓度DATS(0、25、50和100μM)中培养24小时后,或在相同DATS浓度(50μM)下培养四个时间段(24、48、72和96小时)后的形态变化。采用定量实时聚合酶链反应和蛋白质印迹法检测Saos-2细胞中GRP78 mRNA和蛋白质的表达水平。利用小干扰RNA抑制Saos-2细胞中的GRP78表达,随后将这些细胞用于研究DATS处理的抗增殖作用。
由于DATS浓度增加和作用时间延长,GRP78 mRNA和蛋白质的表达水平显著下调(<0.05)。此外,GRP78沉默与DATS处理的细胞增殖之间存在显著关联(<0.05)。
这些结果表明,DATS通过下调GRP78的表达抑制人骨肉瘤Saos-2细胞的生长。