十八碳二烯酸对神经胶质瘤细胞增殖与凋亡的影响及其机制  

Effects of octadecadienoic acid on proliferation and apoptosis of glioma cells and its mechanisms

作　　者：谢明仁 何天晓 袁霞 张璟[2] 俞蕾 俞发荣 XIE Ming-ren;HE Tian-xiao;YUAN Xia;ZHANG Jing;YU Lei;YU Fa-rong(School of Public Security,Gansu University of Political Science and Law,Lanzhou 730070;Gansu Provincial Center for Disease Control and Prevention,Lanzhou 730000,China)

机构地区：[1]甘肃政法大学公安分院,兰州730070 [2]甘肃省疾病预防控制中心,兰州730000

出　　处：《中国应用生理学杂志》2022年第5期434-437,442,共5页Chinese Journal of Applied Physiology

基　　金：甘肃省自然科学金资助项目(21JR7RA571);兰州市科技计划项目(2019-1-48);甘肃政法大学重大项目(GZF2021XZD06)。

摘　　要：目的:探索十八碳二烯酸(ODA)抑制神经胶质瘤细胞增殖与促凋亡作用及其机制。方法:取培养的人神经胶质瘤细胞(细胞密度2×10^(6)cells/L)分为溶剂对照组(给予DMSO,含量为30μl/L)、5-FU组(含量10 mg/L)和十八碳二烯酸组(设0.3、0.6、1.2 mg/L三个剂量组)。用台盼蓝、噻唑蓝(MTT)检测ODA对神经胶质瘤细胞的毒性作用,用酶联免疫吸附法(ELISA)检测神经胶质瘤细胞P53、PI3K、P21、PKB/Akt、caspase-9蛋白表达水平。结果:①光学显微镜细胞计数显示:ODA低、中、高剂量组和5-FU组细胞增殖抑制率比溶剂对照组显著升高(P<0.01),与5-FU组相比差异无统计学意义(P>0.05)。②MTT检测结果显示:与溶剂对照组相比,ODA低、中、高剂量组和5-FU组细胞增殖抑制率显著升高(P<0.01);与5-FU组相比,仅ODA高剂量组细胞增殖抑制率显著增加(P<0.01)。③流式细胞仪检测结果显示:与溶剂对照组相比,ODA低、中、高剂量组和5-FU组G_(0)/G_(1)期细胞数显著增加(P<0.05,P<0.01),G_(2)/M期细胞数显著减少(P<0.01),细胞凋亡率显著增加(P<0.01);与5-FU组相比,仅ODA高剂量组G_(2)/M期细胞数显著减少(P<0.01),凋亡率显著增加(P<0.01)。④ELISA检测结果显示:ODA低、中、高剂量组和5-FU组P53、P13K、PKB/Akt蛋白表达水平比溶剂对照组均显著降低(均P<0.01),仅ODA高剂量组蛋白表达水平均显著低于5-FU组(P<0.01);ODA低、中、高剂量组和5-FU组P21、caspase-9蛋白表达水平明显高于溶剂对照组(P<0.05,P<0.01),仅ODA高剂量组蛋白表达水平显著高于5-FU组(P<0.01)。结论:ODA对神经胶质瘤细胞增殖具有明显的抑制和促凋亡作用,其机制与上调细胞P21、caspase-9水平促凋亡,下调P53、PI3K、PKB/Akt水平抑制细胞分裂周期,降低PI3K-Akt信号转导通路活性有关。Objective:To study the effects of octadecadienoic acid(ODA)on the proliferation and apoptosis of glioma cells and its mechanisms.Methods:Cultured human glioma cells(cell density 2×10^(6)cells/L)were divided into solvent control group(DMSO,30μl/L),5-FU group(10 mg/L)and octadecadienic acid groups(0.3,0.6 and 1.2 mg/L groups).The toxicity of ODA on glioma cells was detected by trypan blue and thiazolium blue(MTT).The expression levels of P53,PI3K,P21,PKB/Akt and Caspase-9 in glioma cells were determined by enzyme-linked immunosorbent assay(ELISA).Results:①Cell count under optical microscope showed that the inhibition rate of cell proliferation in ODA low,medium and high dose groups and 5-FU group was significantly higher than that in the solvent control group(P<0.01),but there was no statistical significance compared with the 5-FU group(P>0.05).②MTT assay showed that the inhibition rate of cell proliferation was increased significantly in ODA low,medium and high dose groups and 5-FU groups(P<0.01),compared with the solvent control group.Compared with 5-FU group,the inhibition rate of cell proliferation was increased significantly only in ODA high dose group(P<0.01).③The number of G_(0)/G_(1)phase cells in ODA low,medium and high dose groups and 5-FU group were increased significantly(P<0.05,P<0.01),the number of G_(2)/M phase cells were decreased significantly(P<0.01),and the apoptosis rate was increased significantly(P<0.01),compared with the solvent control group.Compared with the 5-FU group,the number of cells in G_(2)/M phase was decreased significantly(P<0.01)and the apoptosis rate was increased significantly(P<0.01)in ODA high dose group.④ELISA test results showed that the protein expression levels of P53,PI3K and PKB/Akt in ODA low,medium and high dose groups and 5-FU group were significantly lower than those in solvent control group(all P<0.01),but the protein expression levels in ODA high dose group were significantly lower than those in 5-FU group(P<0.01).The protein expression levels of P21 and

关 键 词：十八碳二烯酸 神经胶质瘤细胞 抑制作用 细胞凋亡 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]