This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Research
Haidong Gong
Corresponding Author
ORCiD: https://orcid.org/0000-0002-2919-0472
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.
Keywords
microRNA-210, glioblastoma, VMP1, proliferation, apoptosis
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
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
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
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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