DADS对人胃癌干样细胞增殖能力及Shh与Gli1表达的影响

Effect of diallyl disulfide on proliferation and Shh and Gli1 expression in human gastric cancer stem like cells

目的观察大蒜有效成分二烯丙基二硫(diallyl disulfide,DADS)对胃癌干样细胞(gastric cancer stem like cells,GCSLCs)增殖能力及干细胞相关分子Shh和Gli1表达的影响,初步探讨其抑制胃癌干样细胞增殖的作用机制。方法采用MTT实验、软琼脂集落形成等方法检测DADS对GCSLCs增殖的影响;采用RT-PCR、Western blot实验观察DADS对GCSLCs中Shh、Gli1在m RNA和蛋白表达水平的作用。结果 MTT实验发现,30mg/L DADS处理48h后GCSLCs组OD值(0.530±0.319)和MGC 803组OD值(0.352±0.027)显著低于未处理GCSLCs组(0.826±0.032)和未处理MGC 803组(0.565±0.023),P<0.05;软琼脂集落形成实验表明,30mg/L DADS处理后GCSLCs组形成的集落小于未处理组,且集落形成数目(15±1.5)也比未处理组(35.5±2.5)少(P<0.05),30mg/LDADS处理MGC 803组形成的集落小于未处理组,且集落形成数目(6.5±0.5)明显少于未处理组(14.5±0.5),P<0.05;RT-PCR实验结果显示,与未处理组比较,30mg/L DADS处理GCSLCs和MGC 803细胞48h后,Shh和Gli1 m RNA的表达水平均显著下调(P<0.05);Western blot结果同样发现,30mg/L DADS处理48h可明显抑制GCSLCs和MGC 803细胞中Shh和Gli1蛋白的表达(P<0.05)。结论 DADS可抑制GCSLCs细胞增殖能力,其作用机制可能与下调Shh和Gli1的表达有关。

Objective To observe the effect on proliferation of gastric cancer stem like cells(GCSLCs) and expression of Shh and Gli1 induced by diallyl disulfide(DADS) and discuss whether its mechanism was related to downregulation of Shh and Gli1 expression in Sonic hedgehog signaling pathway. Methods MTT and soft agar colony formation assay were performed to identify the inhibition effect on proliferation of GCSLCs induced by DADS. RT-PCR and Western blot were conducted to analyse the impact of DADS on m RNA and protein expression of Shh and Gli1 in GCSLCs. Results MTT assay showed that OD value of 30mg/L DADS 48 h treatment groups was notably lower than those of the untreated ones in GCSLCs and MGC 803 cells respectively(P<0.05). The number of colony formation reduced and the sizes of colony formation were far smaller in 30mg/L DADS treatment groups than those in untreatment groups(P<0.05). DADS inhibited proliferation in GCSLCs. DADS inhibited Shh and Gli1 expression in both GCSLCs and MGC 803 cells at m RNA and protein level,compared with untreatment groups respectively(P<0.05). Conclusion DADS induced inhibition proliferation in GCSLCs,which may involve downregulation of Shh and Gli1.