CPVL promotes glioma progression via STAT1 pathway inhibition through interactions with the BTK/p300 axis
Hui Yang, … , Xiuming Liang, Kun Lv
Hui Yang, … , Xiuming Liang, Kun Lv
Published November 16, 2021
Citation Information: JCI Insight. 2021;6(24):e146362. https://doi.org/10.1172/jci.insight.146362.
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Research Article Oncology

CPVL promotes glioma progression via STAT1 pathway inhibition through interactions with the BTK/p300 axis

Abstract

CPVL (carboxypeptidase, vitellogenic-like) is a serine carboxypeptidase that was first characterized in human macrophages. However, the function of CPVL remains unclear in a variety of tumors. The quantitative PCR (qPCR), Western blotting, and IHC assays were utilized to measure the CPVL expression. CPVL was significantly upregulated in glioma cells and tissues compared with normal cells and tissues, respectively. Moreover, high CPVL expression was correlated with advanced clinical grade and poor prognosis. Silencing of CPVL promoted glioma cell apoptosis, and it inhibited cell proliferation and tumorigenicity in vitro and in vivo. Ingenuity Pathway Analysis (IPA) demonstrated that CPVL silencing activated the IFN-γ/STAT1 signaling pathway, thereby inducing glioma cell apoptosis. Mechanistically, immunopurification, mass spectrometry, IP, and glutathione S-transferase (GST) pull-down experiments elucidated that CPVL physically interacts with Bruton’s tyrosine kinase (BTK) and downregulates the STAT1 phosphorylation through promoting p300-mediated STAT1 acetylation. Our findings reveal the crucial role of CPVL in promoting the progression of glioma through suppressing STAT1 phosphorylation. CPVL might serve as a potential prognostic biomarker and therapeutic target for the treatment of glioma.

Authors

Hui Yang, Xiaocen Liu, Xiaolong Zhu, Xueqin Li, Lan Jiang, Min Zhong, Mengying Zhang, Tianbing Chen, Mingzhe Ma, Xiuming Liang, Kun Lv

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Figure 3

CPVL silencing inhibits the proliferation, promotes apoptosis, and regulates cell cycle of glioma cells in vitro.

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CPVL silencing inhibits the proliferation, promotes apoptosis, and regul...
(A and B) Relative CPVL mRNA expression in U251 and LN382 cells expressing CPVL shRNA#1 and CPVL shRNA#2 determined by real-time PCR (n = 3). (C and D) Western blotting analysis of CPVL expression in CPVL-silenced U251 and CPVL-silenced LN382 cells (n = 3). (E and F) MTT assays were used to investigate the cell proliferation rates in CPVL-silenced U251 and CPVL-silenced LN382 cells (n = 3). (G and H) Colony formation assay was used to investigate the cell proliferation capacity of the CPVL-silenced U251 and CPVL-silenced LN382 cells. Representative pictures are shown on the left, and the number of colonies counted are shown on the right (n = 3). (I and J) FACS assay was used to detect the effect of cell apoptosis in CPVL-silenced U251 and CPVL-silenced LN382 cells (n = 3). Representative profiles are shown on the left, and the percentages of cells that were statistically analyzed are shown on the right. (K and L) Cell cycle assays were used to investigate the influence of CPVL silencing on cell cycle in CPVL-silenced U251 and CPVL-silenced LN382 cells (n = 3). The fractions of viable cells in the G1, S, and G2-M phases were quantified by flow cytometry. Representative profiles are shown on the left, and the percentages of cells that were statistically analyzed are shown on the right. All experiments were repeated 3 times. β-Actin was used as a loading control. Bar graph data are presented as mean ± SD. One-way ANOVA with Dunnett’s multiple comparisons test analyses were performed. *P < 0.05.