Abstract
BackgroundRadiotherapy, chemotherapy, and surgery have made crucial strides in glioblastoma treatment, yet they often fail; thus, new treatment and new detection methods are needed. Aberrant expression of Nanos3 has been functionally associated with various cancers. Here, we sought to identify the clinical significance and potential mechanisms of Nanos3 in human glioblastoma.MethodsNanos3 expression was studied in nude mouse glioblastoma tissues and glioblastoma cell lines by immunohistochemistry, Western blot, and RT-PCR. Clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 gene editing assay was performed to generate the Nanos3 knockdown glioblastoma cell lines. The effects of Nanos3 on glioblastoma cells proliferation, migration, invasion, chemoresistance, germ cell characteristics, and tumor formation were analyzed by CCK8, transwell, cell survival experiments and alkaline phosphatase staining in vitro and in nude mouse models in vivo. Correlation between the expression of stemness proteins and the expression of Nanos3 was evaluated by Western blot.ResultsWe found that Nanos3 was strongly expressed in both glioblastoma cell lines and tissues. Western blot and sequencing assays showed that the Nanos3 knockdown glioblastoma cell lines were established successfully, and we discovered that Nanos3 deletion reduced the proliferation, migration, and invasion of glioblastoma cells in vitro (P < 0.05). Nanos3 knockdown enhanced the sensitivity of glioblastoma cells to doxorubicin (DOX) and temozolomide (TMZ) (P < 0.05), and Nanos3+/− glioblastoma cell lines did not show the characteristics of the germline cells. In addition, Nanos3 deletion inhibited subcutaneous xenograft tumor growth in vivo (P < 0.001). Moreover, the oncogenesis germline protein levels of CD133, Oct4, Ki67, and Dazl decreased significantly in glioblastoma cells following Nanos3 knockdown.ConclusionsBoth in vitro and in vivo assays suggest that Nanos3, which is a cancer-germline gene, initiates the tumorigenesis of glioblastoma via acquiring the oncogenesis germline traits. These data demonstrate that ectopic germline traits are necessary for glioblastoma growth.
Highlights
Radiotherapy, chemotherapy, and surgery have made crucial strides in glioblastoma treatment, yet they often fail; new treatment and new detection methods are needed
Nanos3 expression is upregulated in both glioblastoma cell lines and glioblastoma tissues To evaluate whether cancer-germline genes are upregulated in human glioblastoma, Room temperature (RT)-PCR experiments, Western blot assays and immunohistochemistry staining were performed to detect the expression of germlineassociated genes, namely, Nanos3, in human glioblastoma cell lines and glioblastoma tissues (Fig. 1)
RT-PCR showed that Nanos3 was generally overexpressed in glioblastoma multiforme (GBM) cells and the mouse glioblastoma tissues compared with its expression in normal human astrocytes (NHA) cells and the normal mouse brain (Fig. 1a, c)
Summary
Radiotherapy, chemotherapy, and surgery have made crucial strides in glioblastoma treatment, yet they often fail; new treatment and new detection methods are needed. Aberrant expression of Nanos has been functionally associated with various cancers. We sought to identify the clinical significance and potential mechanisms of Nanos in human glioblastoma. The regular care for glioblastoma patients includes surgery followed by radiation and chemotherapy; Dox and TMZ are the conventional. Zhang et al Cancer Cell Int (2020) 20:197 chemotherapeutic drugs for clinical treatment of glioblastoma [4, 5]. Recurrence after regular therapy is unavoidable and results in a high mortality for glioblastoma patients [7, 8]. Therapeutic resistance, recurrence, and poor prognosis have been a global problem for glioblastoma, and little is known about the genetic mechanisms involved in its pathogenesis. Unraveling the mechanism of glioblastoma pathogenesis is essential to the diagnosis, treatment, and prevention of glioblastoma
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