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CDCA7L promotes glioma proliferation by targeting CCND1 and predicts an unfavorable prognosis
Qian-Kun Ji1, Ji-Wei Ma1, Rui-Hua Liu1
1Department of Neurosurgery, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, P.R. China.
Abstract:
Cell division cycle associated 7 like (CDCA7L) belongs to the JPO protein family, recently identified as a target gene of c‑Myc and is frequently dysregulated in multiple cancers. However, to the best of our knowledge, no studies to date have been carried out to investigate the functions of CDCA7L in glioma. Thus, in this study, the expression level of CDCA7L and its association with the prognosis in glioma were detected through the TCGA database. The mRNA expression levels of CDCA7L in glioblastoma (GBM) tissues and normal brain tissues were detected by RT‑qPCR and western blot analysis. To explore the role of CDCA7L in glioma, CDCA7L siRNA was constructed and transfected into U87 glioma cells. The expression levels of CDCA7L and cyclin D1 (CCND1) in glioma U87 cells following transfection with CDCA7L siRNA were measured by RT‑qPCR and western blot analysis. CCK‑8, colony formation, EdU and Transwell assays were used to measure the effects of CDCA7L on U87 cell proliferation, and flow cytometry was used to monitor the changes in the cell cycle following transfection with CDCA7L siRNA. Xenograft tumors were examined in vivo for the carcinogenic effects, as well as the mechanisms and prognostic value of CDCA7L in glioma tissues. The results revealed that CDCA7L was highly expressed in human GBM tissues, and a high expression of CDCA7L was associated with a poor prognosis of glioma patients through the TCGA database. We demonstrated that CDCA7L was highly expressed in human GBM tissues and 3 glioma cell lines. The downregulation CDCA7L expression significantly inhibited the proliferation and colony formation ability of U87 cells by blocking cell cycle progression in the G0/G1 phase. In addition, we found that the mRNA and protein levels of CCND1 were markedly decreased following transfection with CDCA7L siRNA compared with NC siRNA in vitro. The downregulation CDCA7L expression reduced the number of invading cells. Consistent with the results of the in vitro assays, the xenograft assay, immunohistochemistry (IHC) assay and western blot analysis demonstrated that, in response to CDCA7L inhibition, tumor growth was inhibited, Ki‑67 and CCND1 expression levels were decreased in vivo. On the whole, the results of the current study indicate that CDCA7L is highly expressed in human glioma tissues and that a high CDCA7L expression predicts a poor prognosis of glioma patients. CDCA7L promotes glioma U87 cell growth through CCND1.
Insights
Cell division cycle associated 7 like (CDCA7L) is highly expressed in glioma, promoting tumor growth and poor prognosis. Inhibiting CDCA7L reduces proliferation and invasion, offering a potential therapeutic target for this aggressive cancer.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Genetics
Background:
- Cell division cycle associated 7 like (CDCA7L) is a JPO family protein and a target gene of c-Myc, frequently dysregulated in cancers.
- Limited research exists on CDCA7L's function in glioma, necessitating investigation into its role and prognostic significance.
Purpose of the Study:
- To investigate the expression levels and prognostic value of CDCA7L in glioma.
- To explore the functional role of CDCA7L in glioma cell proliferation, cell cycle progression, and invasion.
Main Methods:
- Utilized TCGA database for expression analysis and prognosis correlation.
- Employed RT-qPCR and Western blot for detecting CDCA7L and CCND1 expression in tissues and cell lines.
- Performed in vitro assays (siRNA transfection, CCK-8, colony formation, EdU, Transwell, flow cytometry) and in vivo xenograft models to assess CDCA7L's functional impact.
Main Results:
- CDCA7L is significantly upregulated in glioblastoma (GBM) tissues and glioma cell lines, correlating with poor patient prognosis.
- Downregulation of CDCA7L via siRNA inhibited U87 glioma cell proliferation, colony formation, and invasion by blocking the G0/G1 cell cycle phase.
- CDCA7L inhibition led to decreased expression of cyclin D1 (CCND1) both in vitro and in vivo, with reduced tumor growth and proliferation markers (Ki-67) in xenograft models.
Conclusions:
- CDCA7L is oncogenic in glioma, acting as a potential biomarker for poor prognosis.
- CDCA7L promotes glioma cell growth and invasion, partly through regulating CCND1 expression and cell cycle progression.
- Targeting CDCA7L represents a promising therapeutic strategy for glioma treatment.
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