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Rap2B promotes cell adhesion, proliferation, migration and invasion of human glioma

  • Faan Miao
  • Chenchen Cui
  • Dandan Zuo
  • Hui Zhang
  • Pengjin Mei
  • Hongfu Chen
  • Shuo Wei
  • Fang Yang
  • Junnian Zheng
  • Jin BaiEmail author
  • Yuechao FanEmail author
Laboratory Investigation

Abstract

Purpose

Rap2B, a member of the GTP-binding proteins, is generally up-regulated in numerous types of tumors. Nevertheless, the influence and regulatory mechanisms of Rap2B in gliomas are still not corroborated. Therefore, we analyzed the expression of Rap2B in glioma tissues and cells, and researched its significance in adhesion, proliferation, migration and invasion of the glioma cell line.

Methods

We analyzed the expression of Rap2B in different pathologic grades of glioma tissues by tissue microarray and immunohistochemistry. We assessed the expression of Rap2B in glioma tissue and non-tumor tissue by Western blot. And the expression of Rap2b protein in glioma cells and normal human astrocytes (NHA) was detected by Western blot. In addition, we disclosed the effect of Rap2B knockdown on cell adhesion, proliferation, migration and invasion by using cell attachment assay, CCK-8 assay, cell migration assay and Wound Healing assay, cell invasion assay, respectively. Western blot was used to detect the changes of expression level of NF-kB, MMP-2 and MMP-9 protein when downregulated the expression of Rap2B.

Results

The tissue microarray immunohistochemical results of glioma showed that the expression of Rap2B had no significant correlations between Rap2B expression and the clinicopathologic variables, including patient age (P = 0.352), gender (P = 0.858), WHO Grade (P = 0.693) and histology type (P = 0.877). Western blot analysis showed that the glioma tissue had a dramatically increase of Rap2B expression compared with the non-tumor tissues (P < 0.01). And the expression of Rap2B was markedly up-regulated in all 5 glioma cell lines compared with that in normal human astrocytes (NHA) (P < 0.01). We found that the ability of adhesion, proliferation, migration and invasion of glioma cells were significantly decreased after downregulated Rap2B expression compared with the control group (P < 0.05). In addition, Western blot results showed that the expression levels of NF-kB, MMP-2 and MMP-9 in the interference group were significantly lower than those in the negative control group (P < 0.05).

Conclusions

Rap2B expression is up-regulated in glioma tissues and glioma cell lines. Knockdown of Rap2B inhibits glioma cells’ adhesion and proliferation in vitro. Knockdown of Rap2B inhibits glioma cells’ migration in vitro. Knockdown of Rap2B inhibits glioma cells’ invasion and MMPs activity through NF-kB pathway.

Keywords

Rap2B Glioma Adhesion Proliferation Migration Invasion 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Faan Miao
    • 1
  • Chenchen Cui
    • 1
  • Dandan Zuo
    • 2
  • Hui Zhang
    • 1
  • Pengjin Mei
    • 1
  • Hongfu Chen
    • 1
  • Shuo Wei
    • 1
  • Fang Yang
    • 1
    • 2
    • 3
    • 4
  • Junnian Zheng
    • 3
    • 4
  • Jin Bai
    • 3
    • 4
    Email author
  • Yuechao Fan
    • 1
    Email author
  1. 1.Department of NeurosurgeryThe Affiliated Hospital of Xuzhou Medical UniversityXuzhouPeople’s Republic of China
  2. 2.Department of NeurologyThe Affiliated Hospital of Xuzhou Medical UniversityXuzhouPeople’s Republic of China
  3. 3.Jiangsu Key Laboratory of Biological Cancer TherapyXuzhou Medical UniversityXuzhouPeople’s Republic of China
  4. 4.Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer InstituteXuzhou Medical UniversityXuzhouPeople’s Republic of China

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