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Identification of proteins involved in neural progenitor cell targeting of gliomas.

Staflin K, Zuchner T, Honeth G, Darabi A, Lundberg C - BMC Cancer (2009)

Bottom Line: Moreover, we have developed in vitro assays to mimic the antitumor effect seen in vivo.The addition of antibodies against proteins selected by gene and protein expression analysis either increased or decreased the proliferation rate of the glioma cell lines in vitro.These results suggest that these identified factors might be useful starting points for performing future experiments directed towards a potential therapy against malignant gliomas.

View Article: PubMed Central - HTML - PubMed

Affiliation: CNS Gene Therapy Unit, Dept Experimental Medical Science, Lund University, Lund, Sweden. kstaflin@scripps.edu

ABSTRACT

Background: Glioblastoma are highly aggressive tumors with an average survival time of 12 months with currently available treatment. We have previously shown that specific embryonic neural progenitor cells (NPC) have the potential to target glioma growth in the CNS of rats. The neural progenitor cell treatment can cure approximately 40% of the animals with malignant gliomas with no trace of a tumor burden 6 months after finishing the experiment. Furthermore, the NPCs have been shown to respond to signals from the tumor environment resulting in specific migration towards the tumor. Based on these results we wanted to investigate what factors could influence the growth and progression of gliomas in our rodent model.

Methods: Using microarrays we screened for candidate genes involved in the functional mechanism of tumor inhibition by comparing glioma cell lines to neural progenitor cells with or without anti-tumor activity. The expression of candidate genes was confirmed at RNA level by quantitative RT-PCR and at the protein level by Western blots and immunocytochemistry. Moreover, we have developed in vitro assays to mimic the antitumor effect seen in vivo.

Results: We identified several targets involved in glioma growth and migration, specifically CXCL1, CD81, TPT1, Gas6 and AXL proteins. We further showed that follistatin secretion from the NPC has the potential to decrease tumor proliferation. In vitro co-cultures of NPC and tumor cells resulted in the inhibition of tumor growth. The addition of antibodies against proteins selected by gene and protein expression analysis either increased or decreased the proliferation rate of the glioma cell lines in vitro.

Conclusion: These results suggest that these identified factors might be useful starting points for performing future experiments directed towards a potential therapy against malignant gliomas.

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Validation of protein expression. Immunocytochemical analysis of protein expression of (a-f) Follistatin, (g-l) CD81, (m-r) AXL, Scale bars indicate 50 μm. (s) total protein expression analysis of the proteins Gas6 and CXCL1 comparing gliomas (N25, N29, N32) and NPC (HiB5, ST14A, RN33B). As well as RT-PCR analysis of activin expression in the tumors.
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Figure 3: Validation of protein expression. Immunocytochemical analysis of protein expression of (a-f) Follistatin, (g-l) CD81, (m-r) AXL, Scale bars indicate 50 μm. (s) total protein expression analysis of the proteins Gas6 and CXCL1 comparing gliomas (N25, N29, N32) and NPC (HiB5, ST14A, RN33B). As well as RT-PCR analysis of activin expression in the tumors.

Mentions: Genes highly expressed in the NPC displaying antitumor effects (HiB5/ST14A), according to RNA and protein levels (Figure 3, table 2), were follistatin, Growth arrest specific 6 (Gas6) and ADP-ribosylation factor domain 1 (Arfd1). Expressed genes found in both the tumor group and the NPC with antitumor effect were Camk2, CD81, Kai1, TPT1 and AXL. These genes were not expressed at all or less expressed by the control embryonic cell line RN33B which does not display any antitumor effects. Eef1a was differentially expressed in the different types of tumors and NPC.


Identification of proteins involved in neural progenitor cell targeting of gliomas.

Staflin K, Zuchner T, Honeth G, Darabi A, Lundberg C - BMC Cancer (2009)

Validation of protein expression. Immunocytochemical analysis of protein expression of (a-f) Follistatin, (g-l) CD81, (m-r) AXL, Scale bars indicate 50 μm. (s) total protein expression analysis of the proteins Gas6 and CXCL1 comparing gliomas (N25, N29, N32) and NPC (HiB5, ST14A, RN33B). As well as RT-PCR analysis of activin expression in the tumors.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2713262&req=5

Figure 3: Validation of protein expression. Immunocytochemical analysis of protein expression of (a-f) Follistatin, (g-l) CD81, (m-r) AXL, Scale bars indicate 50 μm. (s) total protein expression analysis of the proteins Gas6 and CXCL1 comparing gliomas (N25, N29, N32) and NPC (HiB5, ST14A, RN33B). As well as RT-PCR analysis of activin expression in the tumors.
Mentions: Genes highly expressed in the NPC displaying antitumor effects (HiB5/ST14A), according to RNA and protein levels (Figure 3, table 2), were follistatin, Growth arrest specific 6 (Gas6) and ADP-ribosylation factor domain 1 (Arfd1). Expressed genes found in both the tumor group and the NPC with antitumor effect were Camk2, CD81, Kai1, TPT1 and AXL. These genes were not expressed at all or less expressed by the control embryonic cell line RN33B which does not display any antitumor effects. Eef1a was differentially expressed in the different types of tumors and NPC.

Bottom Line: Moreover, we have developed in vitro assays to mimic the antitumor effect seen in vivo.The addition of antibodies against proteins selected by gene and protein expression analysis either increased or decreased the proliferation rate of the glioma cell lines in vitro.These results suggest that these identified factors might be useful starting points for performing future experiments directed towards a potential therapy against malignant gliomas.

View Article: PubMed Central - HTML - PubMed

Affiliation: CNS Gene Therapy Unit, Dept Experimental Medical Science, Lund University, Lund, Sweden. kstaflin@scripps.edu

ABSTRACT

Background: Glioblastoma are highly aggressive tumors with an average survival time of 12 months with currently available treatment. We have previously shown that specific embryonic neural progenitor cells (NPC) have the potential to target glioma growth in the CNS of rats. The neural progenitor cell treatment can cure approximately 40% of the animals with malignant gliomas with no trace of a tumor burden 6 months after finishing the experiment. Furthermore, the NPCs have been shown to respond to signals from the tumor environment resulting in specific migration towards the tumor. Based on these results we wanted to investigate what factors could influence the growth and progression of gliomas in our rodent model.

Methods: Using microarrays we screened for candidate genes involved in the functional mechanism of tumor inhibition by comparing glioma cell lines to neural progenitor cells with or without anti-tumor activity. The expression of candidate genes was confirmed at RNA level by quantitative RT-PCR and at the protein level by Western blots and immunocytochemistry. Moreover, we have developed in vitro assays to mimic the antitumor effect seen in vivo.

Results: We identified several targets involved in glioma growth and migration, specifically CXCL1, CD81, TPT1, Gas6 and AXL proteins. We further showed that follistatin secretion from the NPC has the potential to decrease tumor proliferation. In vitro co-cultures of NPC and tumor cells resulted in the inhibition of tumor growth. The addition of antibodies against proteins selected by gene and protein expression analysis either increased or decreased the proliferation rate of the glioma cell lines in vitro.

Conclusion: These results suggest that these identified factors might be useful starting points for performing future experiments directed towards a potential therapy against malignant gliomas.

Show MeSH
Related in: MedlinePlus