摘要
目的 观察高氧、低氧、极低氧微环境中胶质瘤细胞生物学行为变化,并探讨其可能的机制。方法 取人胶质瘤U251细胞进行体外培养,将细胞分为极低氧组、低氧组、常氧组、高氧组,分别置于5%极低氧条件(37℃,5%CO_(2),5%O_(2))、10.5%低氧条件(37℃,5%CO_(2),10.5%O_(2))、21%常氧条件(37℃,5%CO_(2),21%O_(2))、42%高氧条件(37℃,5%CO_(2),42%O_(2))中培养96 h。采用CCK-8检测各组细胞在0、24、48、72、96 h时的增殖情况,流式细胞术检测细胞凋亡情况,Transwell实验检测细胞侵袭能力,实时定量qPCR法检测细胞HIF-1α mRNA表达,DCFH-DA荧光探针染色法检测细胞ROS水平。结果 与常氧组比较,极低氧组、低氧组、高氧组在24、48、72、96 h的增殖能力均降低;低氧组在48、72、96 h的增殖能力高于极低氧组和高氧组(P<0.05或<0.01)。与常氧组比较,极低氧组、低氧组、高氧组细胞凋亡率升高、细胞侵袭数量减少;与低氧组比较,极低氧组和高氧组细胞凋亡率升高、细胞侵袭数量减少(P均<0.05)。与常氧组比较,较极低氧组、低氧组、高氧组细胞HIF-1α mRNA表达水平升高(P均<0.01);与低氧组比较,极低氧组HIF-1α mRNA表达水平升高、高氧组表达水平降低(P<0.05或<0.01)。与常氧组比较,低氧组、极低氧组、高氧组中细胞ROS水平均升高(P均<0.01);与低氧组比较,极低氧组和高氧组细胞ROS水平升高,且极低氧组高于高氧组(P均<0.01)。结论 U251细胞在正常氧微环境中生长状态最佳,低氧和高氧微环境均不利于其增殖和侵袭并能促进其凋亡;这些差异可能与氧浓度改变引发ROS/HIF-1信号通路调节有关。
Objective To observe the changes of biological behavior of glioma cells under different oxygen microen⁃vironment,and to explore its possible mechanism.Methods Human glioma U251 cells were cultured in vitro.The cells were divided into the very hypoxia group,hypoxia group,normoxia group,and hyperoxia group,respectively.The cells were exposed to 5%very hypoxia(37℃,5%CO_(2),5%O_(2)),10.5%hypoxia(37℃,5%CO_(2),10.5%O_(2)),and 21%nor⁃moxia(37℃,5%CO_(2),10.5%O_(2)),and 42%hyperoxia conditions(37℃,5%CO_(2),42%O_(2))for 96 h.CCK-8 was used to detect the proliferation of cells at 0,24,48,72,and 96 h.Flow cytometry was used to detect the apoptosis.Transwell assay was used to detect the invasion ability of cells.HIF-1αmRNA expression was detected by real-time quantitative qP⁃CR.The level of ROS was detected by DCFH-DA fluorescent probe staining.Results Compared with the normoxia group,the proliferation abilities of the very hypoxia group,the hypoxia group and the hyperoxia group decreased at 24,48,72 and 96 h.The proliferation ability of the hypoxia group at 48,72 and 96 h was higher than that of the very hypoxia group and the hyperoxia group(P<0.05 or P<0.01).Compared with the normoxia group,the apoptosis rates of the very hypoxia group,the hypoxia group and the hyperoxia group increased,and the numbers of invasion cells decreased;com⁃pared with the hypoxia group,the apoptosis rates increased and the numbers of invasion cell decreased in the very hypoxia group and the hyperoxia group(all P<0.05).Compared with the normoxia group,the expression levels of HIF-1αmRNA in the cells increased in the very hypoxia group,hypoxia group and hyperoxia group(all P<0.01).Compared with the hy⁃poxia group,the expression levels of HIF-1αmRNA increased in the very hypoxia group and decreased in the hyperoxia group (P<0. 05 or P<0. 01). Compared with the normoxia group, the ROS levels in the hypoxia group, the very hypoxia group and the hyperoxia group increased (all P<0. 01). Compared with the hypoxia group, the levels of
作者
冯鹏程
曹奕强
汤荡
宋海
龙江
王永刚
FENG Pengcheng;CAO Yiqiang;TANG Dang;SONG Hai;LONG Jiang;WANG Yonggang(First Department of Neurosurgery,The First Affiliated Hospital of Kunming Medical University,Kunming 650000,China)
出处
《山东医药》
CAS
2023年第26期44-48,共5页
Shandong Medical Journal
关键词
胶质瘤细胞
氧浓度
肿瘤微环境
细胞凋亡
细胞增殖
glioma cells
oxygen concentration
tumor microenvironment
apoptosis
cell proliferation
