Methylation-associated silencing of miR-9-1 promotes nasopharyngeal carcinoma progression and glycolysis via HK2.

Characteristically, cancer cells metabolize glucose through aerobic glycolysis, known as the Warburg effect. Accumulating evidence suggest that during cancer formation, microRNAs (miRNAs) could regulate such metabolic reprogramming. In the present study, miR-9-1 was identified as significantly hypermethylated in nasopharyngeal carcinoma (NPC) cell lines and clinical tissues. Ectopic expression of miR-9-1 inhibited NPC cell growth and glycolytic metabolism, including reduced glycolysis, by reducing lactate production, glucose uptake, cellular glucose-6-phosphate levels, and ATP generation in vitro and tumor proliferation in vivo. HK2 (encoding hexokinase 2) was identified as a direct target of miR-9-1 using luciferase reporter assays and Western blotting. In NPC cells, hypermethylation regulates miR-9-1 expression and inhibits HK2 translation by directly targeting its 3' untranslated region. MiR-9-1 overexpression markedly reduced HK2 protein levels. Restoration of HK2 expression attenuated the inhibitory effect of miR-9-1 on NPC cell proliferation and glycolysis. Fluorescence in situ hybridization results indicated that miR-9-1 expression was an independent prognostic factor in NPC. Our findings revealed the role of the miR-9-1/HK2 axis in the metabolic reprogramming of NPC, providing a potential therapeutic strategy for NPC.

Gemcitabine plus cisplatin versus fluorouracil plus cisplatin as a first-line concurrent chemotherapy regimen in nasopharyngeal carcinoma: a prospective, multi-institution, randomized controlled phase II study.

BACKGROUND: The objective of this study was to evaluate the efficacy and safety of gemcitabine plus cisplatin concurrent chemoradiotherapy (CCRT) in patients with nasopharyngeal carcinoma. METHOD: Patients with NPC were randomly assigned to the gemcitabine plus cisplatin (GP) group or fluorouracil plus cisplatin (PF) group. Primary end-point was disease-free survival (DFS); secondary endpoints: overall survival, distant metastasis-free survival (DMFS), locoregional relapse-free survival, and treatment-related adverse events. RESULTS: Seventy-six patients were prospectively enrolled and the median follow-up time was 41 months (9-61 months). Three-year DFS were similar between the GP and PF groups (73.7% vs. 60.5%, HR 0.66, 95% CI 0.30-1.44; P = 0.30). Distant metastasis was the most common failure form in PF compared with GP (P = 0.034). Three-year DMFS was significantly better in the GP group than PF group (89.5% vs. 71.1%, P = 0.045). Grade 3-4 gastrointestinal toxicities (vomiting and diarrhea) were significantly more common in the PF group; grade 3-4 neutropenia and thrombocytopenia were more common in the GP group. CONCLUSION: Gemcitabine plus cisplatin could be used as an alternative regimen in CCRT for nasopharyngeal carcinoma.

Candidate tumor suppressor ZNF154 suppresses invasion and metastasis in NPC by inhibiting the EMT via Wnt/ß-catenin signalling.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is especially prevalent in southeast Asia and southern China, but its molecular mechanisms remain poorly characterized. DNA methylation is associated with initiation and progression of tumors, including NPC. Through a genome-wide DNA methylation screening approach, we discovered ZNF154, but its methylation status and roles in NPC have not been investigated. METHODS: The methylation status of ZNF154 in NPC was detected with Methylation specific-PCR (MSP) and Quantitative Sequenom MassARRAY. The invasion and migration capacities were examined by wound healing and transwell invasion assays. The role of ZNF154 in NPC metastasis was clarified with experimental metastasis assay in vivo. Western blotting analysis was used to investigate protein changes followed by ZNF154 over-expression. Kaplan-Meier analysis was performed to determine the association between ZNF154 methylation and prognosis in NPC. RESULTS: Compared to immortalized nasopharyngeal tissues and cells, ZNF154 expression was frequently downregulated in NPC tissues and cell lines due to promoter methylation. Demethylation treatment with 5-aza-2-deoxycytidine (5-Aza) restored ZNF154 expression in NPC cell lines. Ectopic overexpression of ZNF154 in NPC cells inhibited cell migration and invasion in vitro and lung nodule formation in an in vivo tumor metastasis assay. Mechanistic investigations suggested ZNF154 inhibits Wnt/ß-catenin signalling pathway activation and prevents the EMT in NPC. Furthermore, Kaplan-Meier analysis showed hypermethylation of the ZNF154 promoter was associated with significantly poorer disease-free survival (P = 0.032) and distant metastasis-free survival (P = 0.040) among patients with locoregionally advanced NPC. CONCLUSIONS: Taken together, these findings define a novel role for ZNF154 as a tumor suppressor in NPC.

Matched analysis of induction chemotherapy plus chemoradiotherapy versus induction chemotherapy plus radiotherapy alone in locoregionally advanced nasopharyngeal carcinoma: a multicenter study.

BACKGROUND: The relative efficacy of induction chemotherapy (IC) followed by concurrent chemoradiotherapy (CCRT) versus IC followed by radiotherapy (RT) alone in locoregionally advanced NPC remains unclear. METHODS: A total of 877 patients with locally advanced NPC who underwent IC/CCRT or IC/RT at four institutions in China between January 2004 and December 2010 were retrospectively assessed. IC was cisplatin-based combination chemotherapy; concurrent chemotherapy, single agent cisplatin. Two-dimensional conventional radiotherapy (2DCRT) was the radiotherapy technique. All patients were matched in an equal ratio using a pair-matched method. Overall survival (OS), disease-free survival (DFS), distant metastasis-free survival (DMFS), locoregional relapse-free survival (LRRFS) and toxicities were assessed. RESULTS: Eligible patients were matched (n = 642; 321 patients per arm) based on eight clinicopathological characteristics. Five-year OS, DFS, DMFS, and LRRFS were 76%, 70%, 86%, and 88% for IC/CCRT and 75%, 70%, 90%, and 91% for IC/RT, respectively. There were no statistically significant survival differences between arms (P>0.05), even in subgroup analysis. In multivariate analysis, treatment (IC/CCRT vs. IC/RT) was not an independent prognostic factor for any survival end-point. Grade 3/4 acute gastrointestinal toxicities (vomiting, nausea) and hematological toxicities (leucopenia/neutropenia, thrombocytopenia and anemia) were significantly more common in the IC/CCRT arm than IC/RT arm during RT. CONCLUSION: Overall, IC/CCRT failed to demonstrate any survival advantage but higher acute toxicities over IC/RT in locoregionally advanced NPC.

[Comparison of anterior segment changes before and after laser peripheral iridectomy by anterior segment optical coherence tomography in eyes with primary acute angle closure glaucoma].

OBJECTIVE: To evaluate the changes of anterior segment configuration after surgical peripheral iridectomy (SPI) in patients with primary acute angle closure glaucoma (PAACG) by using anterior segment optical coherence tomography (AS-OCT). METHODS: This retrospective self control study consisted of thirty-seven eyes of 37 patients with PAACG who were consecutively recruited in Zhongshan Ophthalmic Center. The peripheral anterior synechiae (PAS) of these patients was less than 5 clock time point. Central anterior chamber depth (ACD), angle opening distance (AOD), trabecular iris area (TISA), angle recess area (ARA), anterior chamber width (ACW), anterior chamber volume (ACV), and crystalline lens rise (CLR) were measured using AS-OCT before and one month after SPI. RESULTS: After SPI, AOD (0.125 ± 0.072) µm, TISA (0.091 ± 0.041) mm(2), ARA (0.095 ± 0.042) mm(2), ACA (14.230 ± 2.000) mm(2) and ACV (90.074 ± 16.796) mm(3) were significantly increased compared with before SPI AOD (0.088 ± 0.078) µm, TISA (0.050 ± 0.048) mm(2), ARA (0.059 ± 0.057) mm(2), ACA (12.332 ± 2.457) mm(2), ACV (73.131 ± 16.976) mm(3) (t = -8.015 to 1.066, P = 0.001 to 0.044), respectively. There were no significantly changes in ACD, ACW and CLR (t = -1.505 to 0.516, P = 0.102 to 0.609). CONCLUSIONS: PAACG can be controlled by SPI resulting in an increase of AOD, TISA, ARA, ACA and ACV, but not ACD or CLR.

Damage patterns of retinal nerve fiber layer in acute and chronic intraocular pressure elevation in primary angle closure glaucoma.

AIM: To observe the differences of damage patterns of retinal nerve fiber layer (RNFL) between acute and chronic intraocular pressure (IOP) elevation in primary angle closure glaucoma (PACG) using optical coherence tomography (OCT). METHODS: Twenty-four patients (48 eyes) with unilateral acute PACG (APACG) attack in the 6 months after admission and 36 patients (64 eyes) with chronic PACG (CPACG) were included in this prospective study. For all cases, IOP has been controlled under 21mmHg after treatment. Using stratus OCT, the RNFL thickness was assessed in eyes with PACG within 3 days, 2 weeks, 1, 3 and 6 months after IOP was controlled. Repeated measures ANOVA was used to examine the changes of RNFL thickness at different time after IOP being controlled in both acute attack eyes and unaffected fellow eyes of APACG and eyes with CPACG. RESULTS: The mean RNFL thickness for the APACG-attacked eyes increased significantly within 3 days (121.49±23.84)µm after acute onset and then became thinner along with time [(107.22±24.72)µm at 2 weeks,(93.58±18.37)µm at 1 month, (84.10±19.89)µm at 3 months and (78.98±19.17)µm at 6 months]. In APACG-attacked eyes, there were significant differences of average RNFL thickness at 5 different times after IOP was controlled (P<0.001). In the APACG unaffected fellow eyes and CPACG eyes, there were no significant differences in mean RNFL thickness at 5 different times(F=0.450, P=0.104 in APACG unaffected fellow eyes and F=1.558, P=0.200 in CPACG eyes). There was significant difference for interaction between time periods and groups (F=1.912, P=0.003). CONCLUSION: RNFL damage patterns are different under different IOP elevated courses. In APACG, RNFL was found to be swollen and thickening right after acute attack and then becomes thinning and atrophy along with the time, while RNFL was found to be diffused thinness in CPACG.

Growth responses of in vitro Fusarium oxysporum f. sp. niveum to external supply of tannic acid.

Allelochemicals released from root exudates or decaying residues of plants play diversified roles in ecological interactions of plant-pathogen. The objective of this work was to evaluate the allelopathic effect of an externally supplied tannic acid on soil-borne in vitro Fusarium oxysporum f. sp. niveum. Results showed that the tannic acid decreased the growth of the fungus up to 9.5% at 800 mg l(-1). Conidial germination was reduced by 52.3% in comparison with the control. However, sporulation and mycotoxin production by the fungus were stimulated. The activity of pectinase and proteinase were initially increased and finally decreased with increase in concentrations of tannic acid. Tannic acid served as an ecological allelochemical, repressing the growth of the pathogen.