Suppressing UVRAG Induces Radiosensitivity by Triggering Lysosomal Membrane Permeabilization in Hypopharyngeal Squamous Cell Carcinoma.

INTRODUCTION: Radiotherapy is one of the most important methods in the treatment of patients with hypopharyngeal squamous cell carcinoma (HSCC). However, radioresistance will be developed after repeated irradiation. Among many key factors contributing to radioresistance, enhanced autophagy is recognized as one of the most important. The ultraviolent irradiation resistance-associated gene (UVRAG) is reported to be a crucial gene involved in the process of autophagy. Here, we test whether UVRAG has effect on the radioresistance of HSCC. METHODS: HSCC cell line Fadu cells were treated with irradiation to test levels of autophagy. Tumor tissues from primary and recurrent HSCC patients were tested by immunohistochemistry. Then, we knocked down UVRAG to test its role in cell growth and the malignant behaviors. Response of cells to treatment was examined using LDH release assay, immunofluorescence, Western blot analysis and colony formation. RESULTS: We found that irradiation induced autophagy in Fadu cells. Immunohistochemistry of primary and irradiated HSCC tumor tissues showed that UVRAG was upregulated after irradiation treatment. Inhibiting UVRAG with siRNA interfered cell growth, cell cycle, malignant behaviors and autophagic flux in Fadu cells. Knocking down UVRAG increased DNA damage and cell death induced by irradiation. Finally, we found that inhibiting UVRAG induced lysosomal membrane permeabilization, which contributed to radiosensitization of Fadu cells. CONCLUSION: Our findings supported the oncogenic properties of UVRAG in HSCC and inhibiting UVRAG increased radiosensitivity in HSCC by triggering lysosomal membrane permeabilization. Therefore, UVRAG might be a promising target in the treatment of HSCC.

Identification of Multiple Cryptococcal Fungicidal Drug Targets by Combined Gene Dosing and Drug Affinity Responsive Target Stability Screening.

UNLABELLED: Cryptococcus neoformans is a pathogenic fungus that is responsible for up to half a million cases of meningitis globally, especially in immunocompromised individuals. Common fungistatic drugs, such as fluconazole, are less toxic for patients but have low efficacy for initial therapy of the disease. Effective therapy against the disease is provided by the fungicidal drug amphotericin B; however, due to its high toxicity and the difficulty in administering its intravenous formulation, it is imperative to find new therapies targeting the fungus. The antiparasitic drug bithionol has been recently identified as having potent fungicidal activity. In this study, we used a combined gene dosing and drug affinity responsive target stability (GD-DARTS) screen as well as protein modeling to identify a common drug binding site of bithionol within multiple NAD-dependent dehydrogenase drug targets. This combination genetic and proteomic method thus provides a powerful method for identifying novel fungicidal drug targets for further development. IMPORTANCE: Cryptococcosis is a neglected fungal meningitis that causes approximately half a million deaths annually. The most effective antifungal agent, amphotericin B, was developed in the 1950s, and no effective medicine has been developed for this disease since that time. A key aspect of amphotericin B's effectiveness is thought to be because of its ability to kill the fungus (fungicidal activity), rather than just stop or slow its growth. The present study utilized a recently identified fungicidal agent, bithionol, to identify potential fungicidal drug targets that can be used in developing modern fungicidal agents. A combined protein and genetic analysis approach was used to identify a class of enzymes, dehydrogenases, that the fungus uses to maintain homeostasis with regard to sugar nutrients. Similarities in the drug target site were found that resulted in simultaneous inhibition and killing of the fungus by bithionol. These studies thus identify a common, multitarget site for antifungal development.

[Anatomical measurements and clinical applications through facial recess approach].

OBJECTIVE: To provide the anatomic data for the correlated otologic microsurgery by the microdissection of temporal bone through facial recess approach. METHOD: Sixteen human temporal bones of eight adult cadaveric heads were dissected under surgical microscope through facial recess approach, and the relative anatomic structures were observed and measured, such as the bony entrance of facial recess approach, facial nerve, stapes, round window, round window niche, pyramidal eminence, cochleariform process, etc. The data were analyzed statistically. RESULT: The width of the bony entrance of facial recess approach was (2.94 +/- 0.32) mm, the height was (8.83 +/- 0.84) mm, the depth was (3.51 +/- 0.17) mm. The distances from stapes to tympanic segment of facial nerve, mastoid segment of facial nerve, round window, cochleariform process and anterior ligament of malleus were (1.38 +/- 0.21) mm, (6.94 +/- 0.47) mm, (3.60 +/- 0.55)mm, (2.23 +/- 0.33)mm, (4.93 +/- 0.61) mm, respectively. The distances from pyramidal eminence to tympanic segment of facial nerve, mastoid segment of facial nerve, round window, round window niche and cochleariform process were (1.05 +/- 0.09) mm, (5.63 +/- 0.41) mm, (3.01 +/- 0.34) mm, (3.29 +/- 0.44) mm, (4.13 +/- 0.51) mm, respectively. The distances from round window to cochleariform process and tympanic segment of facial nerve were (5.11 +/- 0.61) mm and (3.97 +/- 0.61) mm. The distances from round window niche to tympanic segment of facial nerve and mastoid segment of facial nerve were (4.13 +/- 0.38) mm and (7.28 +/- 0.29) mm. CONCLUSION: The facial recess approach played an important role in modern otologic microsurgery. The position of anatomical structure was constant relatively, including short crus of incus, stapes, pyramidal eminence and cochleariform process, etc. These could be used as reference marks for otologic microsurgery.