天麻素对胶质瘤细胞增殖和凋亡的影响

Effect of gastrodin on proliferation and apoptosis of glioma cells

目的 研究天麻素(GAS)对胶质瘤细胞增殖和凋亡的影响及其作用机制。方法 用含体积分数10%胎牛血清的DMEM培养基于37℃、体积分数5%CO;的细胞培养箱中培养T98G细胞,实验分为0,2.5,5.0,10μmol·L^(-1)天麻素组和control组、GAS组、GAS+Vector组、GAS+HOTAIR组。以细胞计数法(CCK-8)试剂盒检测细胞增殖情况;以流式细胞术检测细胞凋亡率情况;以实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测HOTAIR的表达;以蛋白质印迹法检测自噬相关蛋白表达水平。结果 0,2.5,5.0,10μmol·L^(-1)天麻素组培养72 h后细胞OD值分别为1.03±0.10,0.86±0.05,0.72±0.06,0.52±0.09,天麻素可显著抑制T98G细胞增殖,且具有浓度依赖性;0,2.5,5.0,10μmol·L^(-1)天麻素组细胞凋亡率分别为(8.85±1.19)%,(13.57±1.38)%,(17.78±1.72)%,(22.17±1.82)%,天麻素可显著抑制T98G细胞凋亡,且具有浓度依赖性。Control组、GAS组、GAS+Vector组和GAS+HOTAIR组的细胞凋亡率分别为(9.38±1.02)%、(21.16±1.69)%、(21.51±1.20)%和(9.98±1.18)%,HOTAIR表达水平分别为1.00±0.07,0.33±0.03,0.34±0.04,0.78±0.06,天麻素可通过HOTAIR诱导T98G细胞凋亡,可下调HOTAIR表达水平;天麻素可显著下调LC3-Ⅱ/LC3-Ⅰ、Beclin1蛋白表达水平,上调p62蛋白表达水平。GAS组与Control组比较,GAS+HOTAIR组与GAS+Vector组比较,差异均有统计学意义(均P<0.05)。结论 天麻素可以通过长链非编码RNA HOTAIR调节自噬,促进胶质瘤细胞凋亡。

Objective To investigate the effect and mechanism of gastrodin(GAS) on proliferation and apoptosis of glioma cells.MethodsT98G cells were cultured with DMEM containing 10%fetal bovine serum at 37℃in a cell incubator with 5%CO;.The experiment was divided into 0,2.5,5.0,10μmol·L^(-1)gastrodin groups and control group,GAS group,GAS+Vector group,GAS+HOTAIR group.Cell proliferation was detected by cell counting kit-8 (CCK-8).Cell apoptosis rate was detected by flow cytometry.The expression of HOTAIR was detected by real-time quantitutive reverse transcription polymerase chain reaction (qRT-PCR).Western blot was used to detect the expression of autophagy related proteins.Results The OD values of cells cultured for 72 h in 0,2.5,5.0,10μmol·L^(-1)gastrodin groups were 1.03±0.10,0.86±0.05,0.72±0.06 and 0.52±0.09,gastrodin could significantly inhibit T98 G cell proliferation in a concentration-dependent manner.The apoptosis rates of T98G cells in 0,2.5,5.0,10μmol·L^(-1)gastrodin groups were (8.85±1.19)%,(13.57±1.38)%,(17.78±1.72)%,(22.17±1.82)%,gastrodin can significantly inhibited apoptosis of T98G cells in a concentration-dependent manner.The apoptosis rates of control group,GAS group,GAS+Vector group and GAS+HOTAIR group were (9.38±1.02)%,(21.16±1.69)%,(21.51±1.20)%,(9.98±1.18)%,respectively,HOTAIR expression levels were 1.00±0.07,0.33±0.03,0.34±0.04 and0.78±0.06.Gastrodin could induce apoptosis of T98G cells by HOTAIR,gastrodin can down-regulate the expression level of HOTAIR.Gastrodin could significantly down-regulated LC3-Ⅱ/LC3-Ⅰand Beclin1 protein expression levels,and up-regulated p62 protein expression levels.There were significant differences between GAS group and control group,between GAS+HOTAIR group and GAS+Vector group (P<0.05).Conclusion Gastrodin can regulate autophagy and promote apoptosis of glioma cells through long non-coding RNA HOTAIR.