HOXA4调控Wnt/β-catenin信号通路对裸鼠移植胶质瘤的抑制作用

doi:10.13481/j.1671-587x.20190302

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (03): 474-478.doi: 10.13481/j.1671-587x.20190302

• 基础研究 • 上一篇

HOXA4调控Wnt/β-catenin信号通路对裸鼠移植胶质瘤的抑制作用

周海霞¹, 侯力键², 王正明³, 田宇³, 韩亮², 李密馥⁴

1. 吉林大学中日联谊医院特需病房, 吉林长春 130033;
2. 吉林大学中日联谊医院病理科, 吉林长春 130033;
3. 吉林大学中日联谊医院神经外科, 吉林长春 130033;
4. 武警吉林总队医院神经外科, 吉林长春 130052

收稿日期:2018-05-04 发布日期:2019-06-05
通讯作者: 李密馥,副主任医师(Tel:0431-80882430,E-mail:373967458@qq.com);韩亮,主治医师(Tel:0431-89876953,E-mail:hlzr@jlu.edu.cn) E-mail:373967458@qq.com;hlzr@jlu.edu.cn
作者简介:周海霞(1984-),女,吉林省长春市人,主治医师,医学硕士,主要从事肿瘤分子病理学方面的研究。
基金资助:
国家自然科学基金资助课题（30672159）；教育部高校博士学科点专项科研基金资助课题（20110061110070）；吉林省卫计委卫生与健康青年科技骨干培养计划项目资助课题（2018Q025）

Inhibitory effect of HOXA4 on transplantationglioma of nude mice through regulating Wnt/β-catenin signal pathway

ZHOU Haixia¹, HOU Lijian², WANG Zhengming³, TIAN Yu³, HAN Liang², LI Mifu⁴

1. Unit of VIP, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2. Department of Pathology, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
3. Department of Neurosurgery, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
4. Department of Neurosurgery, Armed Police Jilin General Hospital, Changchun 130052, China

Received:2018-05-04 Published:2019-06-05

摘要/Abstract

摘要： 目的：建立人胶质瘤荷瘤裸鼠模型，探讨HOXA4对胶质瘤U251细胞体内生长的影响以及对Wnt/β-catenin信号通路的调控作用及其机制。方法：采用慢病毒转染，建立胶质瘤U251细胞稳转同源异型盒基因A4（HOXA4） siRNA细胞系（si-HOXA4）和稳转空载体阴性对照U251细胞系（si-NC）。取对数生长期U251、si-NC和si-HOXA4细胞接种于BALB/c裸鼠颈背部皮下，建立荷瘤裸鼠模型，命名为对照组、si-NC组和si-HOXA4组。观察各组裸鼠成瘤情况并绘制肿瘤生长曲线；培养21 d处死裸鼠后剥离肿瘤组织，测量肿瘤体积和质量；qRT-PCR法检测各组裸鼠肿瘤组织中HOXA4、CTNNB1和Gsk3β mRNA相对表达量；免疫组织化学（IHC）法检测各组裸鼠肿瘤组织中HOXA4、β-catenin、Gsk3β、CyclinD1和P53蛋白表达水平。结果：si-HOXA4组裸鼠肿瘤体积和质量小于si-NC组和对照组（P<0.05）；si-HOXA4组裸鼠肿瘤组织中HOXA4 mRNA相对表达量和HOXA4蛋白表达水平低于其他2组（P<0.05）；与si-NC组和对照组比较，si-HOXA4组裸鼠肿瘤组织中CTNNB1 mRNA相对表达量降低（P<0.05），β-catenin和CyclinD1蛋白表达水平亦降低（P<0.05），但Gsk3β和P53蛋白表达水平明显升高（P<0.05）。结论：抑制人胶质瘤U251细胞HOXA4表达可在体内通过Wnt/β-catenin信号通路调控CyclinD1和P53蛋白的表达，从而抑制荷瘤裸鼠肿瘤的生长。

关键词: 胶质瘤, 同源异型盒基因A4, Wnt/β-catenin, 信号通路, 裸鼠, 细胞周期蛋白D1, P53蛋白

Abstract: Objective:To establish the glioma-bearing nude mouse models, and to investigate the effect of HOXA4 on the growth of glioma U251 cells in vivo and its regulatory effect on the Wnt/β-catenin signal pathway and its mechanism. Methods:The glioma U251 cell line stably transfected with HOXA4 siRNA(si-HOXA4) and the U251 cell line stably transfected with blank vector (si-NC) were established by lentivirus transfection. The U251, si-NC, and si-HOXA4 cells were respectively inoculated under the skin of the neck and back of the BALB/c nude mice to establish the glioma-bearing nude mouse models named as control group, si-NC group, and si-HOXA4 group. The tumorigenesis of nude mice in various groups were observed and the tumor growth curve was drawn. The tumor tissue was stripped after the mice were sacrificed on the 21 th day, and the volume and weight of tumor were measured; the relative mRNA expression amounts of HOXA4, CTNNB1, and Gsk3β in tumor tissue of the nude mice in various groups were detected by qRT-PCR method; the expression levels of HOXA4, β-catenin, Gsk3β, CyclinD1, and P53 proteins in tumor tissue of the nude mice in various groups were detected by immunohistochemistry (IHC) method. Results:Compared with si-NC group and control group, the volume and weight of tumor of the nude mice in si-HOXA4 group were significantly decreased(P<0.05). The relative expression amount of HOXA4 mRNA and the expression level of HOXA4 protein in si-HOXA4 group were significantly lower than those in the other groups (P<0.05). Compared with si-NC group and control group, the relative expression amount of CTNNB1 mRNA and the expression levels of β-catenin and CyclinD1 proteins in si-HOXA4 group were significantly decreased (P<0.05), and the expression levels of Gsk3β and P53 proteins were significantly increased (P<0.05). Conclusion:Inhibition of HOXA4 expression in human glioma U251 cells can regulate the expressions of CyclinD1 and P53 through Wnt/-catenin signal pathway in vivo, thus inhibiting the tumor growth of glioma-bearing nude mice.

Key words: glioma, homeobox gene A4, Wnt/β-catenin, signal pathway, nude mice, CyclinD1, P53

中图分类号:

R739.4

周海霞, 侯力键, 王正明, 田宇, 韩亮, 李密馥. HOXA4调控Wnt/β-catenin信号通路对裸鼠移植胶质瘤的抑制作用[J]. 吉林大学学报(医学版), 2019, 45(03): 474-478.

ZHOU Haixia, HOU Lijian, WANG Zhengming, TIAN Yu, HAN Liang, LI Mifu. Inhibitory effect of HOXA4 on transplantationglioma of nude mice through regulating Wnt/β-catenin signal pathway[J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 474-478.

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HOXA4调控Wnt/β-catenin信号通路对裸鼠移植胶质瘤的抑制作用

Inhibitory effect of HOXA4 on transplantationglioma of nude mice through regulating Wnt/β-catenin signal pathway

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