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Jinpu Yu
Tianjin Medical University Cancer Institute and Hospital: Tianjin Tumor Hospital
Corresponding Author
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Backgroumd: Tumor metastasis is a process in which tumor cells enter the lymphatic vessels and blood vessels and then spread to the secondary site where they form secondary tumors. In vascular biology, angiogenesis and anti-angiogenesis therapy have been extensively studied, however, the molecular mechanisms involved in lymphangiogenesis and lymphatic metastasis remain unclear.
Methods: We analyzed mRNA expression profiles of 937 primary lung squamous cell carcinoma (LUSC) samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to screen the most differentially expressed genes related to the poor prognosis of LUSC patients and validated in an independent Chinese LUSC cohort. We focused on Vasohibin 2 (VASH2) and investigated its biological functions in LUSC proliferation, apoptosis, migration, invasion, as well as lymphangiogenesis by forced over-expressing VASH2 in LUSC cell line H520 in vitro. We also investigated the anti-tumor efficacy of VASH2 target treatment in LUSC xenograft-bearing mice models.
Results: We identified 12 genes closely related to poor prognosis of LUSC patients, among which VASH2 was validated in an independent Chinese LUSC cohort and displayed high potential of lymphatic metastasis. VASH2 promoted the proliferation and invasion of LUSC cells both in vitro and vivo. Forced over-expression of VASH2 in LUSC cells promoted the amplification and tube-formation of human umbilical vein endothelial cells (HUVECs) and human lymphatic endothelial cells (HLECs) cells via up-regulating vascular endothelial growth factor-D (VEGF-D) production which could be reversed by Snail inhibition. Furthermore, blocking VASH2/VEGF-D signaling using specific antibodies dramatically inhibited tumor growth in mice by interfering proliferation of cancer cells and lymphangiogenesis in tumor tissues.
Conclusion: In conclusion, VASH2 facilitated lymphangiogenesis and tumor growth in a Snail-dependent manner which might serve as a novel biomarker for early diagnosis and prognosis prediction, as well as a potential therapeutic target in LUSC.
Statement of conflict of interest: The authors declare no potential conflicts of interest.
Keywords
Vasohibin 2, Lymphangiogenesis, LUSC, Snail, VEGF-D
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This is a list of supplementary files associated with this preprint. Click to download.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- Table1.pdf
- Table1.pdf
- Table1.pdf
- Table1.pdf
- Table2.pdf
- Table2.pdf
- Table2.pdf
- Table2.pdf
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A new preprint design is coming!
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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
Jinpu Yu
Tianjin Medical University Cancer Institute and Hospital: Tianjin Tumor Hospital
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 15 Dec, 2020
No community comments so far
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Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Backgroumd: Tumor metastasis is a process in which tumor cells enter the lymphatic vessels and blood vessels and then spread to the secondary site where they form secondary tumors. In vascular biology, angiogenesis and anti-angiogenesis therapy have been extensively studied, however, the molecular mechanisms involved in lymphangiogenesis and lymphatic metastasis remain unclear.
Methods: We analyzed mRNA expression profiles of 937 primary lung squamous cell carcinoma (LUSC) samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to screen the most differentially expressed genes related to the poor prognosis of LUSC patients and validated in an independent Chinese LUSC cohort. We focused on Vasohibin 2 (VASH2) and investigated its biological functions in LUSC proliferation, apoptosis, migration, invasion, as well as lymphangiogenesis by forced over-expressing VASH2 in LUSC cell line H520 in vitro. We also investigated the anti-tumor efficacy of VASH2 target treatment in LUSC xenograft-bearing mice models.
Results: We identified 12 genes closely related to poor prognosis of LUSC patients, among which VASH2 was validated in an independent Chinese LUSC cohort and displayed high potential of lymphatic metastasis. VASH2 promoted the proliferation and invasion of LUSC cells both in vitro and vivo. Forced over-expression of VASH2 in LUSC cells promoted the amplification and tube-formation of human umbilical vein endothelial cells (HUVECs) and human lymphatic endothelial cells (HLECs) cells via up-regulating vascular endothelial growth factor-D (VEGF-D) production which could be reversed by Snail inhibition. Furthermore, blocking VASH2/VEGF-D signaling using specific antibodies dramatically inhibited tumor growth in mice by interfering proliferation of cancer cells and lymphangiogenesis in tumor tissues.
Conclusion: In conclusion, VASH2 facilitated lymphangiogenesis and tumor growth in a Snail-dependent manner which might serve as a novel biomarker for early diagnosis and prognosis prediction, as well as a potential therapeutic target in LUSC.
Statement of conflict of interest: The authors declare no potential conflicts of interest.
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This is a list of supplementary files associated with this preprint. Click to download.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S1.pdf
Supplement 1 There are 12 highly differentially expressed genes which significantly correlated with patients’ poor prognosis We collected and analyzed the public datasets were downloaded from the GEO database. And we analyzed the correlation between 12 genes and prognosis in patients with LUSC.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- S2.pdf
Supplement 2 We established H520OV-VASH2, H520OV-VASH2+sh-Snail and H520OV-Snail. A The establishment of H520OV-VASH2 was verified by qPCR and Western blot analysis. B We designed a recombinant lentivirus to knock down Snail in H520OV-VASH2 cells (H520OV-VASH2+sh-Snail). C Snail expression in H520CTRL and H520OV-Snail.
- Table1.pdf
- Table1.pdf
- Table1.pdf
- Table1.pdf
- Table2.pdf
- Table2.pdf
- Table2.pdf
- Table2.pdf
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