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Research
HIF-1α and HIF-2α Mediated PARVB Expression Promotes Tumor Growth and Metastasis in Malignantmelanoma
Chunmeng Wang
Corresponding Author
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This work is licensed under a CC BY 4.0 License. Read Full License
Background
Malignant melanoma is the leading cause of skin cancer-related death. The role of PARVB in malignant melanoma remains unclear. Hypoxia is a hallmark of solid tumors including melanoma. But the regulation role of hypoxia in PARVB expression has not been reported.
Methods
Human malignant melanoma tissues, cell lines and their controls were collected. IHC staining, qRT-PCR and Western blot were performed to reveal the differential PARVB expression. The role of PARVB in tumor growth and metastasis of malignant melanoma was evaluated in vitro and in vivo. The regulation role and mechanism of hypoxia and HIFs in PARVB expression was validated by qRT-PCR, Western blot, ChIP-PCR and Luciferase reporter assays.
Results
PARVB was upregulated in malignant melanoma and correlated with patient survival. OverexpressionofPARVB promoted tumor growth and metastasis of malignant melanoma. Furthermore, hypoxia induced HIF-1α and HIF-2α expression activated PARVB transcription and expression through binding to the specific hypoxia-responsive element (HRE) in the promoter region of PARVB.
Conclusions
In malignant melanoma, Hypoxia induced HIF-1α and HIF-2α expression could directly activate PARVB expression, which further promoted tumor growth and metastasis, inducing poor prognosis. These results indicated that PARVB might be a potential therapeutic target for malignant melanoma.
Keywords
melanoma, PARVB, hypoxia, HIF-1α, HIF-2α
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This is a list of supplementary files associated with this preprint. Click to download.
- Additionalfile1Fig.S1.tif
PARVB is overexpressed in melanoma and associated with poor prognosis (A) Melanoma data set (GSE 3189) visualization showed PARVB is overexpressed in melanoma compared with in normal skin and benign nevi. (B) Melanoma data set (GSE 8401) visualization showed PARVB is overexpressed in melanoma metastasis compared with in primary melanoma. (C, D) TCGA data for melanoma patients showed PARVB high expression group exhibited worse overall survival (C) and progression-free survival (D) than low expression group (calculated in http://www.cbioportal.org/). ** p ≤ 0.01, *** p ≤ 0.001.
- Additionalfile5TableS1.xlsx
- Additionalfile6TableS2.xlsx
- Additionalfile2Fig.S2.tif
PARVB overexpression promotes cell proliferation of human melanoma cells. (A) qRT-PCR and Western blot assays were used to verify the overexpression of PARVB in SK-MEL-28 and A375 cells. (B) Melanoma cell proliferation was detected by CCK8 assay after PARVB overexpression in SK-MEL-28 and A375 cells. (C) Cycle distribution of SK-MEL-28 and A375 cells after detection of PARVB overexpression by flow cytometry analysis. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.
- Additionalfile7TableS3.xlsx
- Additionalfile3Fig.S3.tif
Knockout of PARVB in mouse melanoma cell B16 inhibits proliferation and tumorigenicity. (A, B) PARVB knockout was confirmed via DNA sequencing (A) and Western blot (B) assays. (C) Cell proliferation was detected by CCK8 assay after PARVB knockout in B16 cells. (D-E) The tumorigenicity of melanoma cells was evaluated by tumor volume and weight after PARVB knockout in B16 cells.* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.
- Additionalfile4Fig.S4.tif
Knockdown of PARVB suppresses melanoma cell migration. The effect of PARVB on SK-MEL-28 and A375 cell migration was evaluated using a wound healing assay after transfected PARVBshRNAs or control plasmid.
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This is a preprint. It has not completed peer review.
Research
HIF-1α and HIF-2α Mediated PARVB Expression Promotes Tumor Growth and Metastasis in Malignantmelanoma
Chunmeng Wang
Corresponding Author
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 18 Jun, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Orthopedics
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Malignant melanoma is the leading cause of skin cancer-related death. The role of PARVB in malignant melanoma remains unclear. Hypoxia is a hallmark of solid tumors including melanoma. But the regulation role of hypoxia in PARVB expression has not been reported.
Methods
Human malignant melanoma tissues, cell lines and their controls were collected. IHC staining, qRT-PCR and Western blot were performed to reveal the differential PARVB expression. The role of PARVB in tumor growth and metastasis of malignant melanoma was evaluated in vitro and in vivo. The regulation role and mechanism of hypoxia and HIFs in PARVB expression was validated by qRT-PCR, Western blot, ChIP-PCR and Luciferase reporter assays.
Results
PARVB was upregulated in malignant melanoma and correlated with patient survival. OverexpressionofPARVB promoted tumor growth and metastasis of malignant melanoma. Furthermore, hypoxia induced HIF-1α and HIF-2α expression activated PARVB transcription and expression through binding to the specific hypoxia-responsive element (HRE) in the promoter region of PARVB.
Conclusions
In malignant melanoma, Hypoxia induced HIF-1α and HIF-2α expression could directly activate PARVB expression, which further promoted tumor growth and metastasis, inducing poor prognosis. These results indicated that PARVB might be a potential therapeutic target for malignant melanoma.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6