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Research
Haidong Gong
Corresponding Author
ORCiD: https://orcid.org/0000-0002-2919-0472
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This work is licensed under a CC BY 4.0 License. Read Full License
OBJECTIVE: This study was to investigate whether the expression of vacuole membrane protein 1 (VMP1)was correlated with MicoRNA210 (miR-210) in U87 glioma cell line.
MATERIALS AND METHODS: Plasmid was constructed and transfected into U87 glioma cells.The correlation between VMP1 and miR-210 was observed by the double fluorescence sumei experiment. qRT-PCR and Western blotting were used to detect the expression levels of VMP1 at mRNA and protein level, respectively.MTT assay was utilized to examine the effect of miR-210 on cell proliferation and apoptosis.The transwell chamber assay and plate cloning experiments showed the changes in the rate of cell migration and invasion after cells were transfected with miR-210. Computer SPSS 22.0 software was used to perform t-test and analysis of variance on all experimental dat and results were considered statistically significant when P < 0.05.
Results: The results of the analysis of double luciferase showed that the relative fluorescence intensity of miR-210 and VMP1 in the co-staining group was significantly lower comparing to other experimental groups(P <0.001).qRT-PCR and Western blotting experiments showed that the expression level of VMP1 in miR-210 group was downregulated (P < 0.05).The MTT assay showed that the cell length curve of the miR-210 group was less comparing to the control group (P < 0.05).Transwell experiments showed that the number of cells in the miR-210 group was significantly reduced (P < 0.01).Plate cloning experiments showed no significant difference in the number of cell colonies between the experimental group and the control group (P > 0.05).
Conclusion: VMP1 is a potential target gene of miR-210 in U87 glioma cell line, and provides a new option for molecular targeted therapy of gliomas in the future.
Keywords
microRNA-210, glioblastoma, VMP1, proliferation, apoptosis
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This is a preprint. It has not completed peer review.
Research
Haidong Gong
Corresponding Author
ORCiD: https://orcid.org/0000-0002-2919-0472
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 01 Jun, 2020
No community comments so far
Subject Areas
Oncology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
OBJECTIVE: This study was to investigate whether the expression of vacuole membrane protein 1 (VMP1)was correlated with MicoRNA210 (miR-210) in U87 glioma cell line.
MATERIALS AND METHODS: Plasmid was constructed and transfected into U87 glioma cells.The correlation between VMP1 and miR-210 was observed by the double fluorescence sumei experiment. qRT-PCR and Western blotting were used to detect the expression levels of VMP1 at mRNA and protein level, respectively.MTT assay was utilized to examine the effect of miR-210 on cell proliferation and apoptosis.The transwell chamber assay and plate cloning experiments showed the changes in the rate of cell migration and invasion after cells were transfected with miR-210. Computer SPSS 22.0 software was used to perform t-test and analysis of variance on all experimental dat and results were considered statistically significant when P < 0.05.
Results: The results of the analysis of double luciferase showed that the relative fluorescence intensity of miR-210 and VMP1 in the co-staining group was significantly lower comparing to other experimental groups(P <0.001).qRT-PCR and Western blotting experiments showed that the expression level of VMP1 in miR-210 group was downregulated (P < 0.05).The MTT assay showed that the cell length curve of the miR-210 group was less comparing to the control group (P < 0.05).Transwell experiments showed that the number of cells in the miR-210 group was significantly reduced (P < 0.01).Plate cloning experiments showed no significant difference in the number of cell colonies between the experimental group and the control group (P > 0.05).
Conclusion: VMP1 is a potential target gene of miR-210 in U87 glioma cell line, and provides a new option for molecular targeted therapy of gliomas in the future.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
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