JNK上调HOXB4基因表达对胶质瘤细胞增殖和侵袭的影响

The effect of JNK-induced upregulation of HOXB4 gene expression on the proliferation and invasion of glioma cells

摘要

目的：观察胶质瘤组织和细胞中同源盒基因B4（homeobox gene B4，HOXB4）的表达及其对胶质瘤细胞增殖和侵袭的影响，探究HOXB4表达的上游调控机制。方法：基于中国胶质瘤基因组图谱（Chinese Glioma Genome Atlas，CGGA）数据库，分析胶质瘤患者肿瘤组织中HOXB4的表达水平及其与预后的相关性。通过RT-PCR和Western blot检测U251、U373和U87胶质瘤细胞系中HOXB4的表达。通过CCK-8和Transwell实验评估过表达和沉默HOXB4对U87和U251细胞增殖和侵袭的影响。分别用AMPK、p38 MAPK和JNK的抑制剂处理U87细胞，再用RT-PCR和Western blot检测HOXB4的表达水平，CCK-8和Transwell检测细胞的增殖和侵袭程度。在U87细胞中过表达HOXB4，并给予JNK抑制剂，开展RT-PCR、Western blot、CCK-8和Transwell实验，检测HOXB4表达、细胞增殖及侵袭水平。结果：胶质瘤患者的肿瘤组织中HOXB4高表达，且与肿瘤的恶性程度以及患者不良预后相关。U251、U373和U87细胞系均表达HOXB4，其中以U87细胞表达量最高。过表达HOXB4显著增强U87和U251细胞的增殖和侵袭，沉默HOXB4则明显抑制上述表型。JNK抑制剂可显著下调U87细胞中HOXB4的表达，而AMPK抑制剂和p38 MAPK抑制剂则无明显作用。JNK抑制剂可减弱U87细胞的增殖和侵袭，而HOXB4过表达可拮抗JNK抑制剂的上述作用。结论：胶质瘤细胞中JNK激活可上调HOXB4的表达，从而促进细胞的增殖和侵袭。

Abstract

Objective：To investigate the expression of homeobox gene B4 (HOXB4) in glioma tissues and cells, as well as its effects on glioma cell proliferation and invasion, and to explore the upstream regulatory mechanisms of HOXB4 expression. Methods：Based on the Chinese Glioma Genome Atlas (CGGA) database, the expression level of HOXB4 in glioma tissues and its correlation with patient prognosis were analyzed. RT-PCR and Western blot were used to detect HOXB4 expression in U251, U373, and U87 glioma cell lines. The effects of HOXB4 overexpression and silencing on the proliferation and invasion of U87 and U251 cells were assessed using CCK-8 and Transwell assays. U87 cells were treated with inhibitors of AMPK, p38 MAPK, and JNK. The expression of HOXB4 was detected by RT-PCR and Western blot. The cell proliferation and invasion levels were examined by CCK-8 and Transwell assays. HOXB4 was overexpressed in U87 cells in the presence of a JNK inhibitor, and the HOXB4 expression, cell proliferation and invasion were determined by RT-PCR, Western blot, CCK-8, and Transwell assays. Results： HOXB4 was highly expressed in glioma tissues and was associated with tumor malignancy and poor patient prognosis. HOXB4 was expressed in U251, U373, and U87 cell lines, with the highest expression in U87 cells. Overexpression of HOXB4 obviously enhanced the proliferation and invasion of U87 and U251 cells, whereas silencing HOXB4 markedly inhibited these phenotypes. JNK inhibitor significantly downregulated HOXB4 expression in U87 cells, whereas AMPK inhibitor and p38 MAPK inhibitor had no significant effect. JNK inhibitor attenuated the proliferation and invasion of U87 cells, and HOXB4 overexpression antagonized these effects. Conclusion：JNK activation upregulates HOXB4 expression in glioma cells, thereby promoting cell proliferation and invasion.

关键词

胶质瘤 / HOXB4 / JNK / 增殖 / 侵袭

Key words

glioma / HOXB4 / JNK / proliferation / invasion

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基金资助

国家自然科学基金(81902878)

江苏省大学生创新训练计划(202010312039Y)