NOTCH3: A Potential Predictive Biomarker in Malignant Glioma

Purpose: Malignant gliomas have poor prognosis resulting mainly from high level of cell proliferation, invasion and angiogenesis. This hallmark of glioma makes the surgery difficult and increase the incidence of tumor recurrence and resistance to cancer therapy. Identification of novel biomarkers that are critical elements in glioma pathogenesis may help individualize therapy and predict prognosis. NOTCH signaling pathway is commonly deregulated in glioma. Using Genome-wide exploration, we previously identified amplification at the NOTCH3 locus. This amplification was associated with high level of NOTCH3 transcripts, NOTHC3 protein content and VEGFA transcripts. In our current study, we analyzed the effect of NOTCH3 knockdown on cell proliferation, cell migration and VEGFA expression. 

Methods: NOTCH3 specific shRNA lentivirus was used to knockdown NOTCH3. NOTCH3 knockdown was confirmed using RT-PCR, QReal-time PCR and westernblot. The effect of knockdown on cell proliferation, cell migration and VEGFA expression was analyzed using MTT cell proliferation assay, wound healing assay and QReal-time PCR, respectively. 

Results: Notch3 locus amplification associated with worse outcome compared to tumors with non-amplified locus (P=0.00098, 10 vs 28 months median survival, log-rank test). NOTCH3 knockdown significantly reduced cell proliferation, cell migration and VEGF expression compared to the control.

Conclusion: Our results support NOTCH3 prognostic and predictive biomarker role of NOTCH3 in high grade glioma.