Inhibition of FABP6 Reduces Tumor Cell Invasion and Angiogenesis through the Decrease in MMP-2 and VEGF in Human Glioblastoma Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Immunohistochemical Staining of Human Glioma Specimens
2.2. Cell Culture
2.3. Antibodies
2.4. Stable Expression of shRNAs
2.5. Real-Time Polymerase Chain Reaction (RT-PCR)
2.6. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) Assay
2.7. Cell Migration and Invasion Assays
2.8. Western Blotting
2.9. Tube Formation Assay
2.10. Orthotopic Xenograft Animal Model
2.11. Histological and Immunohistochemical Examination
2.12. Terminal Deoxynucleotidyl Transferase (TdT) dUTP Nick-End Labeling (TUNEL) Assay
2.13. Enzyme-Linked Immunosorbent Assay (ELISA) for VEGF in Condition Medium
2.14. Statistical Analysis
3. Results
3.1. FABP6 Had Higher Expression in Glioma
3.2. Analysis of the Expression of FABP and Establishment of shRNA (shFABP6) Stable Clones in Glioma Cell Lines
3.3. Knockdown of FABP6 Decreased the Migration and Invasion Abilities of Invasion-Related Proteins in the LN229 and U87MG Cells
3.4. Tube Formation of Endothelial Cells Was Attenuated by the Knockdown of FABP6 in Glioma Cells
3.5. The Reduced Phosphorylation of JNK, ERK, and p65 Was Caused by FABP6 Inhibition
3.6. FABP6 Knockdown Combined with TMZ Application Attenuated Tumor Progression in the Orthotropic Xenograft Model
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Species | Brand | Catalog Number |
---|---|---|---|
Cathepsin B | Rb | Abcam | ab125067 |
CD31 | Rb | Abcam | ab28364 |
ERK | Rb | Cell Signaling Technology | 9102s |
FABP6 | Rb | Novus | NBP1-32482 |
FAK | Rb | Cell Signaling Technology | 71433s |
GAPDH | Rb | Cell Signaling Technology | 5174s |
JNK | Rb | Cell Signaling Technology | 9252s |
MLC 2 | Rb | Cell Signaling Technology | 8505s |
MMP2 | Rb | Cell Signaling Technology | 13132s |
MMP9 | Rb | Cell Signaling Technology | 13667s |
P65 | Rb | Cell Signaling Technology | 8242s |
Paxillin | Ms | BD Biosciences | 610051 |
p-ERK | Rb | Cell Signaling Technology | 4377s |
p-FAK (Y397) | Ms | BD Biosciences | 611723 |
p-JNK | Rb | Cell Signaling Technology | 4668s |
p-MLC (Ser19) | Rb | Cell Signaling Technology | 3671 |
p-p65 | Rb | Abcam | Ab185619 |
p-paxillin | Rb | Cell Signaling Technology | 2541s |
TIMP-1 | Rb | Abcam | ab109125 |
TIMP-2 | Ms | Millipore | MAB13446 |
VEGFR1 | Rb | Abcam | ab32152 |
VEGFR2 | Rb | Cell Signaling Technology | 2479s |
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Pai, F.-C.; Huang, H.-W.; Tsai, Y.-L.; Tsai, W.-C.; Cheng, Y.-C.; Chang, H.-H.; Chen, Y. Inhibition of FABP6 Reduces Tumor Cell Invasion and Angiogenesis through the Decrease in MMP-2 and VEGF in Human Glioblastoma Cells. Cells 2021, 10, 2782. https://doi.org/10.3390/cells10102782
Pai F-C, Huang H-W, Tsai Y-L, Tsai W-C, Cheng Y-C, Chang H-H, Chen Y. Inhibition of FABP6 Reduces Tumor Cell Invasion and Angiogenesis through the Decrease in MMP-2 and VEGF in Human Glioblastoma Cells. Cells. 2021; 10(10):2782. https://doi.org/10.3390/cells10102782
Chicago/Turabian StylePai, Feng-Cheng, Hsiang-Wei Huang, Yu-Ling Tsai, Wen-Chiuan Tsai, Yu-Chen Cheng, Hsin-Han Chang, and Ying Chen. 2021. "Inhibition of FABP6 Reduces Tumor Cell Invasion and Angiogenesis through the Decrease in MMP-2 and VEGF in Human Glioblastoma Cells" Cells 10, no. 10: 2782. https://doi.org/10.3390/cells10102782