不良心理应激通过上调乳酸脱氧酶A水平促进胶质瘤增殖和侵袭

发布时间：2018-02-16 04:05

本文关键词： 脑胶质瘤不良心理应激侵袭 LDHA 乳酸代谢　出处：《细胞与分子免疫学杂志》2017年05期 　论文类型：期刊论文

【摘要】：目的探讨乳酸脱氢酶A(LDHA)表达在不良心理应激促进脑胶质瘤细胞增殖及侵袭中的作用。方法将胶质瘤荷瘤裸鼠分为荷瘤组、荷瘤应激组、阴性对照组、短发夹RNA(shRNA)-乳酸脱氧酶A(LDHA)组、shRNA-LDHA应激组。束缚应激4周后,测量各组裸鼠肿瘤大小;利用ELISA检测血清中去甲肾上腺素(NE)、肾上腺素(EPI)的含量;通过比色法检测瘤组织中乳酸的量;Western blot法检测各组瘤组织LDHA蛋白表达。采用CCK-8法和平板克隆形成实验检测NE对胶质瘤LN229细胞增殖的影响;TranswellTM法检测其侵袭能力;实时定量PCR检测LN229细胞LDHA mRNA表达水平;Western blot法检测LDHA、胞外信号调节激酶1/2(ERK1/2)、缺氧诱导因子1α(HIF-1α)蛋白表达;利用荧光素酶报告基因活性检测实验研究NE调节LDHA表达的分子机制。结果与荷瘤组比较,荷瘤应激组肿瘤的大小、EPI、NE、乳酸的含量及LDHA蛋白表达水平均明显增加;而沉默LDHA后,肿瘤的增殖速度、乳酸的含量显著降低;与对照组比较,NE促进LN229细胞LDHA mRNA水平,上调LDHA蛋白表达、ERK1/2和HIF-1α蛋白磷酸化水平,且细胞增殖速度、侵袭能力均增强;荧光素酶报告基因实验证实NE通过HIF-1α上调LDHA的表达。结论不良心理应激通过上调LDHA表达促进胶质瘤增殖和侵袭。
[Abstract]:Objective to investigate the role of lactate dehydrogenase (LDH) expression in promoting the proliferation and invasion of glioma cells under adverse psychological stress. Methods Glioma bearing nude mice were divided into three groups: tumor-bearing group, tumor-bearing stress group and negative control group. ShRNA-LDHA stress group in short hairpin RNA-shRNA-lactic deoxylase group. After 4 weeks of restraint stress, tumor size of nude mice in each group was measured, and serum norepinephrine (NE) and epinephrine (EPI) contents were detected by ELISA. The expression of LDHA protein in tumor tissues was detected by colorimetric assay, and the effect of NE on the proliferation of glioma LN229 cells was detected by CCK-8 assay and flat clone formation assay. The invasive ability of NE on the proliferation of glioma LN229 cells was detected by Transwell TM method. The expression level of LDHA mRNA in LN229 cells was detected by real-time quantitative PCR. The expression of extracellular signal-regulated kinase 1 / 2 ERK1 / 2, hypoxia inducible factor-1 伪 (HIF-1 伪) protein was detected by Western blot assay. The molecular mechanism of NE regulating LDHA expression was studied by using luciferase reporter gene activity test. Results compared with the tumor bearing group, the tumor size, lactic acid content and LDHA protein expression level in the tumor-bearing stress group were significantly increased, but after LDHA was silenced, Compared with the control group, the proliferation rate and lactic acid content of the tumor were significantly decreased, the level of LDHA mRNA and the expression of LDHA protein, ERK1 / 2 and HIF-1 伪 were up-regulated in LN229 cells, and the proliferation rate and invasiveness of the cells were enhanced. Luciferase reporter gene experiment confirmed that NE upregulated the expression of LDHA through HIF-1 伪. Conclusion adverse psychological stress can promote the proliferation and invasion of gliomas by up-regulating the expression of LDHA.
【作者单位】：潍坊医学院生物化学教研室;
【基金】：国家自然科学基金(81500798) 山东省自然科学基金(ZR2014JL050) 山东省医药卫生科技发展计划(2015WSB27014)
【分类号】：R739.41

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