CRM197 in Combination With shRNA Interference of VCAM-1 Displays Enhanced Inhibitory Effects on Human Glioblastoma Cells.

Abstract

CRM197 is a naturally nontoxic diphtheria toxin mutant that binds and inhibits heparin-binding epidermal growth factor-like growth factor. CRM197 serves as carrier protein for vaccine and other therapeutic agents. CRM197 also inhibits the growth, migration, invasion, and induces apoptosis in various tumors. Vascular cell adhesion molecule-1 (VCAM-1) is an important cell surface adhesion molecule associated with malignancy of gliomas. In this work, we aimed to investigate the role and mechanism of CRM197 combined with shRNA interference of VCAM-1 (shRNA-VCAM-1) on the migration, invasion, and apoptosis of glioblastoma cells. U87 and U251 human glioblastoma cells were treated with CRM197 (10 µg/ml) and shRNA interfering technology was employed to silence VCAM-1 expression. Cell viability, migration, invasiveness, and apoptosis were assessed with CCK8, Transwell and Annexin V-PE/7-AAD staining. Activation of cleaved caspase-3, 8, and 9, activity of matrix metalloproteinase-2/9 (MMP-2/9), and expression of phosphorylated Akt (p-Akt) were also checked. Results showed that CRM197 and shRNA-VCAM-1 not only significantly inhibited the cell proliferation, migration, invasion, but also promoted the apoptosis of U87 and U251 cells. Combined treatment of both displayed enhanced inhibitory effects on the malignant biological behavior of glioma cells. The activation of cleaved caspase-3, 8, 9 was promoted, activity of MMP-2 and MMP-9 and expression of p-Akt were inhibited significantly by the treatment of CRM197 and shRNA-VCAM-1 alone or in combination, indicating that the combination of CRM197 with shRNA-VCAM-1 additively inhibited the malignant behavior of human glioblastoma cells via activating caspase-3, 8, 9 as well as inhibiting MMP-2, MMP-9, and Akt pathway.

• Authors: Yi Hu, Xing Lin, Ping Wang, Yi-Xue Xue, Zhen Li, Li-Bo Liu, Bo Yu, Tian-Da Feng, Yun-Hui Liu
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 230 Issue 8 Pg. 1713-28 (Aug 2015) ISSN: 1097-4652 [Electronic] United States
• PMID: 25201410 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 Wiley Periodicals, Inc.

Chemical References

• Antineoplastic Agents
• Bacterial Proteins
• RNA, Small Interfering
• Vascular Cell Adhesion Molecule-1
• CRM197 (non-toxic variant of diphtheria toxin)

Topics

• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Bacterial Proteins (pharmacology)
• Blotting, Western
• Cell Line, Tumor
• Cell Movement (drug effects)
• Combined Modality Therapy (methods)
• Genetic Therapy (methods)
• Glioblastoma (pathology)
• Humans
• RNA Interference
• RNA, Small Interfering
• Reverse Transcriptase Polymerase Chain Reaction
• Transfection
• Vascular Cell Adhesion Molecule-1 (metabolism)