Targeting EZH2 depletes LMP1-induced activated regulatory T cells enhancing antitumor immunity in nasopharyngeal carcinoma

Objective: Regulatory T cells (Tregs) are critical factors that impair antitumor immunity. Epstein–Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) is one of the most pathogenic factors in nasopharyngeal carcinoma (NPC). However, the role of EBV-encoded LMP1 in regulating Treg generation in NPC remains unclear. Materials and Methods: The in vitro stability of activated Tregs (aTregs) influenced by LMP1 was analyzed by flow cytometry. The inhibitory effects of LMP1-HONE1 antigen-induced aTregs on tumor-associated antigen (TAA)-specific T cells were analyzed in vitro and in vivo. Finally, the expression of LMP1, Foxp3, and enhancer of zeste homolog 2 (EZH2) were analyzed in samples from 86 NPC patients by immunohistochemistry and immunofluorescence. Results: LMP1 upregulated the expression of EZH2, which increased the stability of aTregs in vitro. EZH2 inhibitor, DZnep, depleted LMP1-HONE1 antigen-induced aTregs in vitro and led to potent TAA-specific T cell antitumor immunity in vivo. In NPC tissues, LMP1 expression level was positively correlated with the number of EZH2+ Tregs, which was positively correlated with clinical stage and overall survival. Conclusions: EZH2 is essential for maintaining the stability and inhibitory functions of aTregs that are induced by EBV-encoded LMP1 in NPC

Lateral endoscopic transorbital approach to middle skull base: anatomical and surgical practice study / 中华耳鼻咽喉头颈外科杂志

Objective@#To investigate the anatomical and surgical approaches to middle cranial fossa through orbital lateral wall under endoscope.@*Methods@#Cadaveric formalin fixed specimens and fresh colored silicone injected specimens were used for this study. All anatomic technical measurements were performed under 0° and 30° endoscope and infrared rays navigation. The surgical approach was designed with the bony opening on the lateral wall of orbit through which the lateral side of the middle cranial fossa could be directly entered under endoscope. One case of recurrent meningioma was performed through this surgical approach. SPSS 20.0 software was used for statistical analysis.@*Results@#The approach can directly enter the middle cranial fossa and expose anatomic landmarks including superior orbital fissure, lateral side of cavernous sinus, foramen rotundum, foramen ovale, foramen spinosum, petrosal bone and others as well as Ⅱ, Ⅲ, Ⅳ, Ⅴ, Ⅵ cranial nerves under endoscope. The maximal bony opening through the lateral wall of orbit was measured, with a horizontal diameter of (1.38±0.68) cm, a vertical diameter of (2.02±0.32) cm, a depth of (1.44±0.42) cm from bony opening margin to the dura. The recurrent meningioma involving lateral side of the middle skull base was successfully removed by this surgical approach through lateral wall of orbit.@*Conclusion@#Lateral transorbital endoscopic approach to the lateral side of middle skull base is a safe, feasible, and minimally invasive method, which allows surgeons to directly manipulate diseases involving this area with good visualization and minimal invasion under endoscope.

Clinical application of supraclavicular flap for oncologic reconstruction of hypopharynx and upper esophagus / 中华耳鼻咽喉头颈外科杂志

Objective@#To assess the efficacy of supraclavicular artery island flap (SCAIF) for the reconstruction of hypopharynx and upper esophagus.@*Methods@#The SCAIF procedure on was used in 10 patients, including 8 with hypopharygeal carcinomas, 1 with esophageal carcinoma and neck skin invasion and 1 with hypopharyngeal leiomyosarcoma, at the Otorhinolaryngology Hospital, the First Affiliated Hospital, Sun Yatsen University between December 2015 and June 2017. The sizes of the flaps were measured in (4-8) cm×(5-12) cm. Clinical indexes such as harvesting time and survival were recorded.@*Results@#Harvesting time for SCAIF ranged from 20 to 30 minutes, averaging 26 minutes. Nine flaps survived, one flap had partially necrosis. Functional outcomes were excellent and the donor sites were direct closed without complications.@*Conclusions@#SCAIF is a versatile, reliable, and easily harvested flap, with good cosmetic and functional outcomes for reconstructing the defects of hypopharynx and upper esophagus.

Inhibition of miR-9 expression suppresses proliferation, invasion and mi-gration of nasopharyngeal carcinoma cells / 中国病理生理杂志

AIM:To investigate the effects of down-regulated miR-9 expression on the proliferation , invasion and migration of nasopharyngeal carcinoma (NPC) cells.METHODS:Human NPC CNE1 and CNE2 cells were transfect-ed with the inhibitor of miR-9 by Lipofectamine to down-regulate the expression of miR-9, and the cells transfected with an inhibitor control were also set up .The cell proliferation and cell cycle were evaluated by CCK-8 assay and flow cytometry . The cell invasion and migration abilities were detected by Transwell invasion and wound -healing assays .Immunoblotting was applied to analyze the levels of the proteins .RESULTS:Compared with control group , inhibition of miR-9 expression in the NPC cells by transfection of the miR-9 inhibitor significantly decreased the proliferation ability (P<0.05).The per-centages of the cells in G 0/G1 phase [ CNE2: ( 57.96 ±1.39 )% vs ( 47.93 ±1.76 )%, P<0.05; CNE1: ( 51.24 ± 0.88)%vs (48.29 ±0.39)%, P<0.05] were significantly increased.The migration distances [CNE2: (186.50 ± 7.94)μm vs (247.56 ±15.56)μm, P<0.05;CNE1:(139.06 ±16.73)μm vs (230.66 ±14.27)μm, P<0.01] and the invasion ability of the CNE2 cells (43.00 ±3.17 vs 65.80 ±5.20, P<0.01) were also significantly inhibited .Moreo-ver, the tumor cells transfected with the inhibitors produced lower β-catenin.CONCLUSION:Inhibition of miR-9 expres-sion suppresses the proliferation , invasion and migration of nasopharyngeal carcinoma cells .