Plasma Epstein-Barr Virus MicroRNA BART8-3p as a Diagnostic and Prognostic Biomarker in Nasopharyngeal Carcinoma

Abstract

Background: Nasopharyngeal carcinoma is an Epstein-Barr virus (EBV)-associated tumor that is highly common in southern China. Our previous sequencing data demonstrated that the EBV-encoded microRNA BART8-3p was most upregulated in nasopharyngeal carcinoma (NPC) and was closely associated with the metastasis of NPC. However, the values of plasma BART8-3p in NPC patients have not yet been well characterized.

Material and methods: We quantified plasma BART8-3p expression by quantitative real-time PCR in 205 newly diagnosed NPC patients. Kaplan-Meier analysis was used to compare overall survival (OS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) between the groups.

Results: Plasma pretreatment BART8-3p was highly expressed in NPC patients compared with healthy controls. Pretreatment BART8-3p yielded a 92% predictive value for detecting NPC. Importantly, BART8-3p decreased dramatically after therapy relative to pretreatment levels. High levels of pretreatment or post-treatment BART8-3p were associated with worse OS, DMFS, and LRRFS. Multivariate analysis showed that high pretreatment or post-treatment BART8-3p was an independent unfavorable prognostic marker for OS (HR 3.82, 95% CI 1.77-8.24, P = .001 or HR 2.74, 95% CI 1.27-5.91, P = .010), DMFS (HR 2.82, 95% CI 1.36-5.85, P = .005 or HR 3.27, 95% CI 1.57-6.81, P = .002), and LRRFS (HR 1.94, 95% CI 1.12-3.35, P = .018 or HR 2.03, 95% CI 1.14-3.62, P = .016) in NPC. Subgroup analysis revealed that for patients with locally advanced NPC with high levels of pretreatment BART8-3p (n = 58), more cycles of chemotherapy (≥6 cycles) tended to prolong OS (P = .070). Over 50% (6/11) patients with high levels of post-treatment BART8-3p presented distant metastasis.

Conclusion: Plasma BART8-3p is a promising biomarker for the detection and prognosis of NPC.

Keywords: BART8-3p; Epstein-Barr virus; biomarker; metastasis; nasopharyngeal carcinoma.

Figure 1.

Plasma BART8-3p in healthy controls (HCs) and NPC patients. (A) BART8-3p expression in NPC tissues in another microRNA microarray (GSE36682). (B, C) Levels of plasma BART8-3p in HCs and NPC patients. (D, E, F) Levels of BART8-3p in different NPC tumor stages, node stages and TNM stages. ∗ P < .05, ∗∗ P < .01 and ∗∗∗ P < .001.

Figure 2.

Clinical application of plasma BART8-3p in NPC. (A) ROC curve analysis of BART8-3p for discriminating NPC from healthy controls. (B) Correlation of BART8-3p and EBV DNA load among patients with NPC. (C) Change in BART8-3p in NPC patients after therapy. (D) Evolution of BART8-3p in a locally advanced NPC patient who developed liver metastasis 10 months after completing the entire therapy. ∗∗∗ P < .001.

Figure 3.

Kaplan-Meier estimates of overall survival (OS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) according to pretreatment or post-treatment BART8-3p in 205 NPC patents. Comparison of OS (A), DMFS (B), and LRRFS (C) according to BART8-3p pretreatment (upper row). Comparison of OS (D), DMFS (E), and LRRFS (F) according to BART8-3p post-treatment (lower row).

Figure 4.

Kaplan-Meier estimates of overall survival (OS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) according to pretreatment or post-treatment BART8-3p in 173 locally advanced NPC patents from 205 NPC patients. Comparison of OS (A), DMFS (B), and LRRFS (C) according to pretreatment with BART8-3p (top row). (D, E, F) Comparison of OS, DMFS and LRRFS according to post-treatment BART8-3p (middle row). Comparison of OS (G), DMFS (H), and LRRFS (I) in 58 locally advanced NPC patients with high levels of pretreatment BART8-3p (bottom row).