姜黄素作用于miR-539对胃癌细胞生物学行为影响的研究

Effect of Curcumin on Biological Behavior of Gastric Cancer Cells Through miR-539

背景:研究发现,姜黄素可调控多种miRNAs表达来影响肿瘤细胞的生物学行为,而姜黄素是否通过调控miR-539表达影响胃癌细胞的生物学行为尚未明确。目的:探讨姜黄素通过调控miR-539表达对胃癌细胞增殖、迁移、侵袭、凋亡等生物学行为的影响及其作用机制。方法:收集49例胃癌患者组织,将胃癌AGS、SGC7901细胞分为空白组、姜黄素组、阴性对照组、miR-539 mimics组、miR-539抑制剂组、抑制剂对照组、miR-539抑制剂+姜黄素组。采用qRT-PCR法检测胃癌组织和细胞中miR-539 mRNA表达。MTT法检测细胞增殖,划痕实验和Transwell实验检测细胞迁移和侵袭,流式细胞术检测细胞凋亡,蛋白质印迹法检测APOBEC3B、c-Myc、cyclin D1、claudin-1、N-cadherin蛋白表达。结果:胃癌组织和细胞中miR-539表达均显著低于对照组(P<0.05),且miR-539表达与肿瘤分期有关(P<0.05)。姜黄素可剂量依赖性地上调胃癌细胞中miR-539表达(P<0.05)。与相应对照组相比,miR-539 mimics组、姜黄素组胃癌细胞增殖、迁移、侵袭能力均明显降低,细胞凋亡率明显增加,APOBEC3B、c-Myc、cyclin D1、claudin-1、N-cadherin蛋白表达明显降低(P<0.05)。结论:MiR-539在胃癌组织中表达降低,姜黄素可通过上调miR-539表达来抑制胃癌细胞增殖、迁移、侵袭,并诱导细胞凋亡。

Background:Studies have shown that curcumin can regulate the expressions of a variety of miRNAs and affect tumor cell biological behavior.However,whether curcumin affects the biological behavior of gastric cancer cells by regulating expression of miR-539 has not been clarified.Aims:To investigate the effect and mechanism of curcumin on proliferation,migration,invasion,apoptosis of gastric cancer cells by regulating the expression of miR-539.Methods:A total of 49 gastric cancer tissue specimens were collected.Gastric cancer AGS,SGC7901 cells were divided into blank group,curcumin group,negative control group,miR-539 mimics group,miR-539 inhibitor group,inhibitor control group,miR-539 inhibitor+curcumin group.qRT-PCR was used to detect the mRNA expression of miR-539 in gastric cancer tissue and cells.Cell proliferation was detected by MTT.Scratch test,Transwell test were used to detected cell migration and invasion.Cell apoptosis was determined by flow cytometry.Western blotting was used to detect protein expressions of APOBEC3B,c-Myc,cyclin D1,claudin-1 and N-cadherin.Results:Expressions of miR-539 mRNA in gastric cancer tissue and gastric cancer cells were significantly decreased than those in corresponding controls(P<0.05),and was related to tumor stage in gastric cancer patients(P<0.05).Curcumin up-regulated the expression of miR-539 mRNA in a dose-dependent manner(P<0.05).Compared with corresponding control group,cell proliferation,migration and invasion were significantly decreased in miR-539 mimics group and curcumin group(P<0.05),cell apoptosis rate was significantly increased(P<0.05),and protein expressions of APOBEC3B,c-Myc,cyclin D1,claudin-1 and N-cadherin were significantly decreased(P<0.05).Conclusions:Expression of miR-539 is decreased in gastric cancer,and curcumin can inhibit proliferation,migration and invasion of gastric cancer cells and induce cell apoptosis by up-regulating the expression of miR-539.