摘要
目的探讨同源盒基因A7(HOXA7)在脑胶质瘤组织的表达及其对胶质瘤细胞增殖和凋亡的影响。方法选择2010年9月至2016年8月新乡医学院第一附属医院神经外科手术切除的46例神经胶质瘤标本和10例颅脑外伤手术切除的正常脑组织,应用实时荧光定量聚合酶链式反应检测脑胶质瘤组织和正常脑组织中HOXA7 mRNA相对表达量。选取对数生长期U251细胞随机分为空白对照组、无义序列对照组和HOXA7 siRNA组,空白对照组细胞不进行转染,无义序列对照组细胞转染无义序列Scrambled小干扰RNA(siRNA),HOXA7 siRNA组细胞转染HOXA7 siRNA;应用实时荧光定量聚合酶链式反应检测3组细胞中HOXA7 mRNA相对表达量,细胞计数盒-8增殖实验检测3组细胞增殖活力,流式细胞仪检测3组细胞的细胞周期及凋亡率。结果高级别脑胶质瘤中HOXA7的mRNA相对表达量显著高于低级别脑胶质瘤和正常脑组织,低级别脑胶质瘤中HOXA7 mRNA相对表达量显著高于正常脑组织(P<0.05)。HOXA7 siRNA组U251细胞中HOXA7 mRNA相对表达量显著低于空白对照组和无义序列对照组(P<0.05),空白对照组与无义序列对照组U251细胞中HOXA7 mRNA相对表达量比较差异无统计学意义(P>0.05)。培养24、48、72、96 h时,HOXA7 siRNA组U251细胞增殖活力显著高于空白对照组和无义序列对照组(P<0.001);空白对照组与无义序列对照组U251细胞增殖活力比较差异无统计学意义(P>0.05)。HOXA7 siRNA组G_(0)/G_(1)期U251细胞占比显著高于空白对照组和无义序列对照组(P<0.05);空白对照组与无义序列对照组G_(0)/G_(1)期U251细胞占比比较差异无统计学意义(P>0.05)。HOXA7 siRNA组S期U251细胞占比显著低于空白对照组和无义序列对照组(P<0.05);空白对照组和无义序列对照组S期U251细胞占比比较差异无统计学意义(P>0.05)。HOXA7 siRNA组G_(2)/M期U251细胞占比显著高于空白对照组和无义序列对照组(P<0.05);空白对照组与无义序列对照组G_(2)/M期U251细胞占比比较差异无统计学意义(P>0.05)。HOXA7 siRNA组U251细胞凋亡率显著高于空白对照组和无义序列对照组(P<0.05),空白对照组与无义序列对照组U251细胞凋亡率比较差异无统计学意义(P>0.05)。结论HOXA7在脑胶质瘤组织中呈高表达,且随着脑胶质瘤级别的增加其表达水平显著增高;HOXA7可能通过促进脑胶质瘤细胞的增殖能力和抑制脑胶质瘤细胞的凋亡,参与脑胶质瘤的发生发展。
Objective To explore the expression of homeobox gene-A7(HOXA7)in glioma tissue and its effect on proliferation and apoptosis of glioma cells.Methods A total of 46 glioma specimens removed during neurosurgery and 10 normal brain tissues surgically removed from craniocerebral trauma in the Department of Neurosurgery of the First Affiliated Hospital of Xinxiang Medical University from September 2010 to August 2016 were selected.The relative expression of HOXA7 mRNA in glioma tissue and normal brain tissue was examined by real-time quantitative polymerase chain reaction.U251 cells in the logarithmic growth phase were randomly divided into the blank control group,the nonsense sequence control group and the HOXA7 siRNA group.The U251 cells in the blank control group were not transfected,the U251 cells in the nonsense sequence control group were transfected with scrambled small interfering RNA(siRNA),and the U251 cells in the HOXA7 siRNA group were transfected with HOXA7 siRNA.The expression of HOXA7 mRNA in U251 cells in the three groups was measured by using the real-time quantitative polymerase chain reaction,the proliferation activity of U251 cells in the three groups was detected by using the cell counting kit-8 assay,and the cell cycle and apoptosis rate of U251 cells in the three groups were detected by using the flow cytometry.Results The relative expression of HOXA7 mRNA in high-grade glioma was significantly higher than that in the low-grade glioma and normal brain tissue,and the relative expression of HOXA7 mRNA in low-grade glioma was significantly higher than that in normal brain tissue(P<0.05).The relative expression of HOXA7 mRNA in U251 cells in the HOXA7 siRNA group was significantly lower than that in the blank control group and the nonsense sequence control group(P<0.05).There was no statistically significant difference in the relative expression of HOXA7 mRNA in U251 cells between the blank control group and the nonsense sequence control group(P>0.05).At 24,48,72,and 96 hours of culture,the proliferation activity of U251 cells in the HOXA7 siRNA group was significantly higher than that in the blank control group and the nonsense sequence control group(P<0.01);and there was no significant difference in the proliferation activity of U251 cells between the blank control group and the nonsense sequence control group(P>0.05).The proportion of U251 cells in the G_(0)/G_(1) phase in the HOXA7 siRNA group was significantly higher than that in the blank control group and the nonsense sequence control group(P<0.05),and there was no significant difference in the proportion of U251 cells in the G_(0)/G_(1) phase between the blank control group and the nonsense sequence control group(P>0.05).The proportion of U251 cells in the S phase in the HOXA7 siRNA group was significantly lower than that in the blank control group and the nonsense sequence control group(P<0.05),and there was no significant difference in the proportion of U251 cells in S phase between the blank control group and the nonsense sequence control group(P>0.05).The proportion of U251 cells in the G_(2)/M phase in the HOXA7 siRNA group was significantly higher than that in the blank control group and the nonsense sequence control group(P<0.05),and there was no significant difference in the proportion of U251 cells in the G_(2)/M phase between the blank control group and the nonsense sequence control group(P>0.05).The apoptosis rate of U251 cells in the HOXA7 siRNA group was significantly higher than that in the blank control group and the nonsense sequence control group(P<0.05),and there was no significant difference in the apoptosis rate of U251 cells between the blank control group and the nonsense sequence control group(P>0.05).Conclusion HOXA7 is highly expressed in glioma tissues,and its expression significantly increases with the glioma grade.HOXA7 may be involved in the occurrence and development of glioma by promoting the proliferation of glioma cells and inhibiting the apoptosis of glioma cells.
作者
张志永
周祥
汲乾坤
周文科
金保哲
ZHANG Zhiyong;ZHOU Xiang;JI Qiankun;ZHOU Wenken;JIN Baozhe(Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical University/Henan Key Laboratory of Neural Rehabilitation,Weihui 453100,Henan Province,China)
出处
《新乡医学院学报》
CAS
2024年第7期645-650,共6页
Journal of Xinxiang Medical University
基金
国家自然科学基金资助项目(编号:81541030)
河南省科技攻关计划项目(编号:162102310120)。
