Serglycin-Mediated Selective Reactivation of the Novel YAP/CRISPLD2 Developmental Axis Promotes Metastasis and Sorafenib Resistance in Hepatocellular Carcinoma
Abstract Background: The extracellular matrix (ECM) component serglycin promotes the epithelial-to-mesenchymal transition (EMT) and metastasis in an autocrine manner. However, the detailed mechanism underlying the roles of serglycin in Hepatocellular carcinoma (HCC) metastasis progression remains to be clarified.Methods: Analyzing 123 patients’ serum by ELISA to investigate the association between serglycin expression and HCC metastasis and prognosis. Serglycin, CRISPLD2, and YAP were overexpressed or knocked down with vector or RNAi. The RNA - sequence was used to screen serglycin downstream effectors, followed by bioinformatics, ChIP-PCR, luciferase, and promoter site mutation to identify novel target genes. The metastatic abilities of serglycin were demonstrated by a series of in vitro and in vivo experiments. Immunofluorescence, flow cytometry, and western blotting were carried out to demonstrate the potential mechanisms of serglycin.Results: We observed that serglycin was overexpressed in HCC tissues and serum, and its level was associated with metastasis and poor prognosis. By gain- and loss-of-function approaches, we found serglycin affects migratory motility and metastatic capacity in vitro and in vivo, and it's significantly associated with the stemness-like properties. Interestingly, serglycin activated the effector YAP of Hippo pathway, and further study verified the crucial role of serglycin/YAP in tumorigenesis by the DEN-induced HCC mouse model. Furthermore, the CRISPLD2 was selectively upregulated by serglycin, the axis of serglycin / YAP / CRISPLD2 regulated metastatic capacity in HCC cells, it revealed CRISPLD2 was the direct target gene of serglycin-mediated YAP/TEAD1 complex. Besides, serglycin-mediated YAP pathway enhanced cell sorafenib resistance, sorafenib combined with verteporfin reversed serglycin-mediated cellular insensitivity to sorafenib therapy.Conclusions: Autocrine ECM serglycin plays a crucial role in the process of stemness maintenance, tumorigenesis, and metastasis via selective reactivation of the novel YAP/CRISPLD2 developmental axis promotes metastasis and sorafenib resistance in Hepatocellular carcinoma, YAP-TEAD1 inhibitor verteporfin reversed serglycin-mediated cellular sorafenib resistance..
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Preprint| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
ResearchSquare.com - (2020) vom: 29. Dez. Zur Gesamtaufnahme - year:2020 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
HU, HAO [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.21203/rs.3.rs-135613/v1 |
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| PPN (Katalog-ID): |
XRA034704647 |
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| 245 | 1 | 0 | |a Serglycin-Mediated Selective Reactivation of the Novel YAP/CRISPLD2 Developmental Axis Promotes Metastasis and Sorafenib Resistance in Hepatocellular Carcinoma |
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| 520 | |a Abstract Background: The extracellular matrix (ECM) component serglycin promotes the epithelial-to-mesenchymal transition (EMT) and metastasis in an autocrine manner. However, the detailed mechanism underlying the roles of serglycin in Hepatocellular carcinoma (HCC) metastasis progression remains to be clarified.Methods: Analyzing 123 patients’ serum by ELISA to investigate the association between serglycin expression and HCC metastasis and prognosis. Serglycin, CRISPLD2, and YAP were overexpressed or knocked down with vector or RNAi. The RNA - sequence was used to screen serglycin downstream effectors, followed by bioinformatics, ChIP-PCR, luciferase, and promoter site mutation to identify novel target genes. The metastatic abilities of serglycin were demonstrated by a series of in vitro and in vivo experiments. Immunofluorescence, flow cytometry, and western blotting were carried out to demonstrate the potential mechanisms of serglycin.Results: We observed that serglycin was overexpressed in HCC tissues and serum, and its level was associated with metastasis and poor prognosis. By gain- and loss-of-function approaches, we found serglycin affects migratory motility and metastatic capacity in vitro and in vivo, and it's significantly associated with the stemness-like properties. Interestingly, serglycin activated the effector YAP of Hippo pathway, and further study verified the crucial role of serglycin/YAP in tumorigenesis by the DEN-induced HCC mouse model. Furthermore, the CRISPLD2 was selectively upregulated by serglycin, the axis of serglycin / YAP / CRISPLD2 regulated metastatic capacity in HCC cells, it revealed CRISPLD2 was the direct target gene of serglycin-mediated YAP/TEAD1 complex. Besides, serglycin-mediated YAP pathway enhanced cell sorafenib resistance, sorafenib combined with verteporfin reversed serglycin-mediated cellular insensitivity to sorafenib therapy.Conclusions: Autocrine ECM serglycin plays a crucial role in the process of stemness maintenance, tumorigenesis, and metastasis via selective reactivation of the novel YAP/CRISPLD2 developmental axis promotes metastasis and sorafenib resistance in Hepatocellular carcinoma, YAP-TEAD1 inhibitor verteporfin reversed serglycin-mediated cellular sorafenib resistance. | ||
| 700 | 1 | |a Xu, Liang |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Tie-Jun |e verfasserin |4 aut | |
| 700 | 1 | |a Luo, Fei-Fei |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Jing |e verfasserin |4 aut | |
| 700 | 1 | |a Mei, Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Zheng, Li-Sheng |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Li-Xia |e verfasserin |4 aut | |
| 700 | 1 | |a Meng, Dong-Fang |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Zhan-Fei |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Shi-Jun |e verfasserin |4 aut | |
| 700 | 1 | |a Qian, Chao-Nan |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Bi-Jun |e verfasserin |4 aut | |
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