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Cited by 16Hepatocellular Carcinoma Treatment and PrognosisHippo pathway signaling and YAP/TAZCancer Cells and Metastasis

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The function of Piezo1 in hepatoblastoma metastasis and its potential transduction mechanism

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Abstract

Background: Hepatoblastoma (HB) is the most common primary malignant liver tumor in children. The prognosis of HB metastasis is poor, despite the increasing diversity of treatment. Piezo, a ubiquitously expressed membrane mechano-transduction protein, is involved in the process of tumor cell migration. Under the gene expression profiling interactive analysis (GEPIA) database, Piezo1 was highly expressed in HB and negatively correlated with the overall survival time. Methods: Firstly, the expression of Piezo1 in both paracancerous and HB tissues (n = 7) was detected, and the prognostic value of Piezo1 was assessed in HB (n = 160) patients. Secondly, the inhibition and overexpression of Piezo1were executed in two HB cell lines, HepG2 and Huh 6. Methyl thiazolyl tetrazolium (MTT), wound healing and trans-well assays were performed to identify the effect of Piezo1 on the proliferation and metastasis of HB cells, respectively. In addition, a co-immunoprecipitation assay was performed to determine whether Piezo1 has an interaction with HIF-1α. Finally, the expressions level of Piezo1, HIF-1α, and VEGF by overexpression/inhibition each other were detected by RT-qPCR and western blots to find a possible signaling channel in HB metastasis. Results: We found that Piezo1 was highly expressed in HB tissues and associated with poor prognosis of patients. Piezo1 was related to cell proliferation in HepG2 and Huh 6 cells. We also found that Piezo1 stimulated HIF-1α expression. Meanwhile, overexpression of Piezo1 promoted the migration and invasion of HB cells, while the promotion was not detected when HIF-1α was suppressed. Additionally, the silencing of HIF-1α inhibited the expression of VEGF, but showed no effect on Piezo1 expression. Conclusion: In this study, we identified that Piezo1 was involved in HB metastasis, and the Piezo1-HIF-1α-VEGF axis could be a possible signaling pathway in HB metastasis.

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Crossref · 10.1016/j.heliyon.2022.e10301

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Figures

Figure 1 High Piezo1 expression in HB tissues was associated with clinical prognosis. A, Piezo1 expression of LIHC tissues by GEPIA database, red represents cancer samples and grey represents normal samples. B, Overall survival analysis of LIHC with high Piezo1 expression by GEPIA database. C-E, Western blot and RT-qPCR analysis of Piezo1 expression in 7 pairs of paracancerous and HB tissues. F, Representative immunohistochemistry images of paracancerous and HB tissues (scale bar = 200 μm). ∗∗P < 0.01 vs Paracancerous, Data were shown as mean ± SD, statistical significance was calculated by student's t-test, one-way ANOVA and two-way ANOVA, respectively.
Figure 1 High Piezo1 expression in HB tissues was associated with clinical prognosis. A, Piezo1 expression of LIHC tissues by GEPIA database, red represents cancer samples and grey represents normal samples. B, Overall survival analysis of LIHC with high Piezo1 expression by GEPIA database. C-E, Western blot and RT-qPCR analysis of Piezo1 expression in 7 pairs of paracancerous and HB tissues. F, Representative immunohistochemistry images of paracancerous and HB tissues (scale bar = 200 μm). ∗∗P < 0.01 vs Paracancerous, Data were shown as mean ± SD, statistical significance was calculated by student's t-test, one-way ANOVA and two-way ANOVA, respectively.CC BY-NC-ND · PubMed Central
Figure 2 Piezo1 induced HB cells proliferation. A-C, Western blot and RT-qPCR analysis of Piezo1 expression after transfected with two si-Piezo1. D, MTT assay of two HB cells with different concentrations of Yoda 1. E, Representative images and the statistical result of colony formation in two HB cells after transfected with si-Piezo1 and Yoda1. ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.
Figure 2 Piezo1 induced HB cells proliferation. A-C, Western blot and RT-qPCR analysis of Piezo1 expression after transfected with two si-Piezo1. D, MTT assay of two HB cells with different concentrations of Yoda 1. E, Representative images and the statistical result of colony formation in two HB cells after transfected with si-Piezo1 and Yoda1. ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.CC BY-NC-ND · PubMed Central
Figure 3 The effect of Piezo1 in HB cells metastasis. A-B, Wound-healing representative images and statistical results of Piezo1 in two HB cells (scale bar = 200 μm). C-D, Transwell representative images and statistical results of Piezo1 on cell invasion (scale bar = 100 μm). ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.
Figure 3 The effect of Piezo1 in HB cells metastasis. A-B, Wound-healing representative images and statistical results of Piezo1 in two HB cells (scale bar = 200 μm). C-D, Transwell representative images and statistical results of Piezo1 on cell invasion (scale bar = 100 μm). ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.CC BY-NC-ND · PubMed Central
Figure 4 Piezo1 promoted HB cells invasion via up-regulation of HIF-1α. A-B, Western blot, and RT-qPCR analysis of Piezo1 and HIF-1α expression in two HB cells after Piezo1 overexpression and inhibition. C-D, Western blot, and RT-qPCR analysis of the inhibition efficiency of GFP-PURO–HIF–1α in two HB cells. E, Trans-well representative images and statistical results of two HB cells after being transfected with si-Piezo1, Yoda 1, GFP-PURO–HIF–1α, and Yoda1+ GFP-PURO–HIF–1α (scale bar = 200 μm). F, co-immunoprecipitation images of HIF-1α and Piezo1. ∗P < 0.05, ∗∗P < 0.01 vs control, #P < 0.05 vs GFP-PURO-control, &&P < 0.01 vs Yoda1, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.
Figure 4 Piezo1 promoted HB cells invasion via up-regulation of HIF-1α. A-B, Western blot, and RT-qPCR analysis of Piezo1 and HIF-1α expression in two HB cells after Piezo1 overexpression and inhibition. C-D, Western blot, and RT-qPCR analysis of the inhibition efficiency of GFP-PURO–HIF–1α in two HB cells. E, Trans-well representative images and statistical results of two HB cells after being transfected with si-Piezo1, Yoda 1, GFP-PURO–HIF–1α, and Yoda1+ GFP-PURO–HIF–1α (scale bar = 200 μm). F, co-immunoprecipitation images of HIF-1α and Piezo1. ∗P < 0.05, ∗∗P < 0.01 vs control, #P < 0.05 vs GFP-PURO-control, &&P < 0.01 vs Yoda1, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.CC BY-NC-ND · PubMed Central
Figure 5 The expression of Piezo1, HIF-1α, and VEGF in two HB cells after transfected with Yoda 1, si-Piezo1 and GFP-PURO–HIF–1α. A-B, Western blot and RT-qPCR analysis of Piezo1 and HIF-1α expression after Piezo1 overexpression and inhibition in two HB cells. C-D, Western blot and RT-qPCR analysis of Piezo1, HIF-1α and VEGF after transfected with si-Piezo1 or GFP-PURO–HIF–1α in two HB cells. E-F, Western blot and RT-qPCR analysis of Piezo1, HIF-1α and VEGF expression after transfected with si-Piezo1 and GFP-PURO–HIF–1α in two HB cells. ∗P < 0.05, ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.
Figure 5 The expression of Piezo1, HIF-1α, and VEGF in two HB cells after transfected with Yoda 1, si-Piezo1 and GFP-PURO–HIF–1α. A-B, Western blot and RT-qPCR analysis of Piezo1 and HIF-1α expression after Piezo1 overexpression and inhibition in two HB cells. C-D, Western blot and RT-qPCR analysis of Piezo1, HIF-1α and VEGF after transfected with si-Piezo1 or GFP-PURO–HIF–1α in two HB cells. E-F, Western blot and RT-qPCR analysis of Piezo1, HIF-1α and VEGF expression after transfected with si-Piezo1 and GFP-PURO–HIF–1α in two HB cells. ∗P < 0.05, ∗∗P < 0.01 vs control, Data were shown as mean ± SD, statistical significance was calculated by one-way ANOVA.CC BY-NC-ND · PubMed Central
FIG4B-Pizeo.tif
FIG4B-Pizeo.tif CC BY-NC-ND · PubMed Central
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Fig5F-HIF-2.tif CC BY-NC-ND · PubMed Central
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Fig5B-HIF1-1.tif CC BY-NC-ND · PubMed Central
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Fig5F-VEGF-1.tif CC BY-NC-ND · PubMed Central
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FIG4F-HIF1-2.tif CC BY-NC-ND · PubMed Central
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Fig1C-GAPDH.tif CC BY-NC-ND · PubMed Central
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FIG4F-Pizeo-1.tif CC BY-NC-ND · PubMed Central
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Fig5F-HIF-1.tif CC BY-NC-ND · PubMed Central
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Fig4F-Pizeo-3.tif CC BY-NC-ND · PubMed Central
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Fig4D-HIF1.tif CC BY-NC-ND · PubMed Central
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Fig-2B-GAPDH.tif CC BY-NC-ND · PubMed Central
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Fig5F-VEGF-2.tif CC BY-NC-ND · PubMed Central
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Fig5B-GAPDH-2.tif CC BY-NC-ND · PubMed Central
FIG4F-Pizeo-2.tif
FIG4F-Pizeo-2.tif CC BY-NC-ND · PubMed Central
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Fig5D-VEGF.tif CC BY-NC-ND · PubMed Central
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Fig5D-HIF-2.tif CC BY-NC-ND · PubMed Central
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Fig5B-HIF-2.tif CC BY-NC-ND · PubMed Central
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Fig5D-HIF-1.tif CC BY-NC-ND · PubMed Central
Fig5F-Pizeo-1.tif
Fig5F-Pizeo-1.tif CC BY-NC-ND · PubMed Central
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fIG5B-Pizeo-1.tif CC BY-NC-ND · PubMed Central
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Fig1C-Pizeo-1.tif CC BY-NC-ND · PubMed Central
FIG4B-HIF1.tif
FIG4B-HIF1.tif CC BY-NC-ND · PubMed Central
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Fig4D-GAPDH.tif CC BY-NC-ND · PubMed Central
Fig5D-Pizeo-1.tif
Fig5D-Pizeo-1.tif CC BY-NC-ND · PubMed Central
Fig2B-piezo-1.tif
Fig2B-piezo-1.tif CC BY-NC-ND · PubMed Central
FIG4F-HIF1.tif
FIG4F-HIF1.tif CC BY-NC-ND · PubMed Central
Fig-5B-Pizeo-2.tif
Fig-5B-Pizeo-2.tif CC BY-NC-ND · PubMed Central
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Fig5D-GAPDH-2.tif CC BY-NC-ND · PubMed Central
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Fig5D-Pizeo-2.tifCC BY-NC-ND · PubMed Central
Fig5F-GAPDH.tif
Fig5F-GAPDH.tif CC BY-NC-ND · PubMed Central
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FIG4F-GAPDH.tif CC BY-NC-ND · PubMed Central
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Fig5B-GAPDH-1.tif CC BY-NC-ND · PubMed Central
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Fig5D-GAPDH-1.tif CC BY-NC-ND · PubMed Central
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Fig4B-GAPDH.tif CC BY-NC-ND · PubMed Central

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Metadata

Authors
Ye, X., Xia, Y., Zheng, Y., Chen, W., Chen, Z., Cheng, Z., & Wang, B.
Affiliations
Shenzhen Children's Hospital (CN)
Corresponding author
Wang, Bin
Venue
Heliyon
Publisher
Elsevier BV
Published
2022-09
Volume
8
Issue
9
Pages
e10301
ISSN
2405-8440 (ISSN-L)
OpenAlex
W4294361000
Cited by
15
References
28
Citations by year
2026: 3 · 2025: 8 · 2024: 3 · 2023: 1
FWCI
1.79
Type
journal article
Language
en
Open access
Locations
  • Heliyon (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Natural Science Foundation of China (81770512, SZSM201812055)
Topics
  • Health Sciences › Medicine › Hepatology › Hepatocellular Carcinoma Treatment and Prognosis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hippo pathway signaling and YAP/TAZ
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
Keywords
Piezo1, hepatoblastoma, metastasis, HIF-1α, vascular endothelial growth factor, mechanotransduction, cell migration and invasion, cell proliferation, prognostic factors

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