Downregulation of KCNMA1 in mice accelerates auditory hair cells senescence via ferroptosis.

KCNMA1 encodes the K+ potassium channel α-subunit that plays a significant role in the auditory system. Our previous studies indicated that KCNMA1 is associated with age-related hearing loss(AHL). However, the detailed mechanism of KCNMA1 involvement in auditory age-related degradation has not been fully clarified. Therefore, we explored the expression of KCNMA1 in the peripheral auditory of 2-month-old and 12-month-old mice by Western blotting and immunofluorescence. The results of animal experiments showed that KCNMA1 expression was decreased in 12-month-old mice compared with 2-month-old mice, whereas the ferroptosis level was increased. To verify the role of KCNMA1 in AHL, we downregulated KCNMA1 in HEI-OC1 cells by transfecting shRNA. After downregulation, the ferroptosis level was increased and the aging process was accelerated. Furthermore, the aging process was affected by the expression of ferroptosis. In conclusion, these results revealed that KCNMA1 is associated with the aging process in auditory hair cells by regulating ferroptosis, which deepens our understanding of age-related hearing loss.

[Evaluation of the efficacy of endoscopic posterior nasal neurectomy with pharyngeal neurectomy of allergic rhinitis combined with chronic rhinosinusitis with nasal polyps].

Objective:To investigate the effect of posterior nasal neurectomy（PNN） with pharyngeal neurectomy （PN） on chronic sinusitis with nasal polyps （CRSwNP）complicated with perennial allergic rhinitis （PAR）. Methods:83 patients with perennial allergic rhinitis combined with chronic group-wide sinusitis with nasal polyps who attended our hospital from July 2020 to July 2021 were selected. All patients underwent conventional functional endoscopic sinusitis surgery（FESS）+ nasal polypectomy. Patients were divided according to whether they underwent PNN+PN. 38 cases in the experimental group underwent FESS combined with PNN+PN; 44 cases in the control group underwent conventional FESS alone. All patients underwent the VAS, RQLQ, and MLK before treatment, and at 6 months and 1 year after surgery. Meanwhile, other relevant data were collected and the preoperative and postoperative follow-up data were collected and analyzed to assess the differences between the two groups. Results:The total postoperative follow-up period was 1 year. The recurrence rate of nasal polyps at 1 year postoperatively and the nasal congestion VAS score at 6 months postoperatively were not statistically significant in the two groups（P>0.05）. However, the patients in the experimental group had statistically significantly lower effusion and sneezing VAS scores, MLK endoscopy scores and RQLQ scores at 6 months and 1 year postoperatively, and nasal congestion VAS scores at 1 year postoperatively compared to the control group（P<0.05）. Conclusion:For patients with perennial AR complicated with CRSwNP, the combination of the PNN+PN in FESS can significantly improve the short-term curative effect, and PNN+PN is a safe and effective surgical treatment.

Increased expression of ST2 on regulatory T cells is associated with cancer associated fibroblast-derived IL-33 in laryngeal cancer.

Interleukin (IL)- 33 plays an essential role in regulatory T cell (Treg)-mediated immunosuppression in cancers and underlies the crosstalk between Tregs and the tumor microenvironment. However, the phenotypic characteristics of subset Tregs modulated by IL-33 and its association with the tumor microenvironment are not fully understood. This study aimed to examine the expression of ST2, the receptor of IL-33, on Tregs in tumors and to evaluate their association with cancer associated fibroblasts (CAFs) and reciprocal influences on the prognosis of laryngeal cancer. Our results showed that increased numbers of Tregs were found in laryngeal tumor tissues. Tregs in stromal IL-33-positive tumor tissues demonstrated significantly higher expression of ST2 than those in IL-33- or adjacent nontumor tissues. ST2-expressing Tregs exhibited upregulation of Ki67 and CTLA4 compared with their ST2- negative counterparts. Furthermore, IL-33 in the tumor microenvironment was mainly derived from fibroblasts. ST2 expression on Tregs was correlated with the number of IL-33-positive CAFs. High ST2 expression on Tregs, combined high ST2 on Tregs and the presence of IL-33 expressing CAFs was associated with worse survival outcomes in laryngeal cancer. This study indicated that increased expression of ST2 on Tregs is associated with microenvironmental IL-33 signaling derived from CAFs in laryngeal cancer, unraveling the special role of Tregs and fibroblasts in modulating IL-33/ST2 involved immune-evasive tumor microenvironment.

Caffeine Induces Autophagy and Apoptosis in Auditory Hair Cells via the SGK1/HIF-1α Pathway.

Caffeine is being increasingly used in daily life, such as in drinks, cosmetics, and medicine. Caffeine is known as a mild stimulant of the central nervous system, which is also closely related to neurologic disease. However, it is unknown whether caffeine causes hearing loss, and there is great interest in determining the effect of caffeine in cochlear hair cells. First, we explored the difference in auditory brainstem response (ABR), organ of Corti, stria vascularis, and spiral ganglion neurons between the control and caffeine-treated groups of C57BL/6 mice. RNA sequencing was conducted to profile mRNA expression differences in the cochlea of control and caffeine-treated mice. A CCK-8 assay was used to evaluate the approximate concentration of caffeine. Flow cytometry, TUNEL assay, immunocytochemistry, qRT-PCR, and Western blotting were performed to detect the effects of SGK1 in HEI-OC1 cells and basilar membranes. In vivo research showed that 120 mg/ kg caffeine injection caused hearing loss by damaging the organ of Corti, stria vascularis, and spiral ganglion neurons. RNA-seq results suggested that SGK1 might play a vital role in ototoxicity. To confirm our observations in vitro, we used the HEI-OC1 cell line, a cochlear hair cell-like cell line, to investigate the role of caffeine in hearing loss. The results of flow cytometry, TUNEL assay, immunocytochemistry, qRT-PCR, and Western blotting showed that caffeine caused autophagy and apoptosis via SGK1 pathway. We verified the interaction between SGK1 and HIF-1α by co-IP. To confirm the role of SGK1 and HIF-1α, GSK650394 was used as an inhibitor of SGK1 and CoCl2 was used as an inducer of HIF-1α. Western blot analysis suggested that GSK650394 and CoCl2 relieved the caffeine-induced apoptosis and autophagy. Together, these results indicated that caffeine induces autophagy and apoptosis in auditory hair cells via the SGK1/HIF-1α pathway, suggesting that caffeine may cause hearing loss. Additionally, our findings provided new insights into ototoxic drugs, demonstrating that SGK1 and its downstream pathways may be potential therapeutic targets for hearing research at the molecular level.

Nicotine induced ototoxicity in rat cochlear organotypic cultures.

Epidemiological evidence has shown that smoking is associated with an increased risk of hearing loss. However, the underlying mechanisms regarding the impact of nicotine on the cochlea remain unclear. This study aimed to investigate the cytotoxic effects of nicotine on cochlear cells using cultured cochlear basilar membranes. Cochlear basilar membranes were isolated from newborn rats, cultured, and treated with 1-100 ng/mL nicotine for 48 h. Cuticular plates and stereocilia bundle staining were used to evaluate hair cell (HC) loss. Spiral ganglion neuron and acoustic nerve fiber staining were assessed to evaluate cochlear neural injury. Scanning electron microscopy and transmission electron microscopy imaging were employed to examine cochlear ultrastructural changes. Our results showed that compared to spiral ganglia and nerve fibers, HCs are more susceptible to nicotine-induced toxicity. HC loss was more severe in the basal turn than in the middle and apical turns, while nerve fibers and spiral ganglion cells were morphologically maintained. Ultrastructural changes revealed disordered and damaged stereocilia, swelling and decreased mitochondrial density, swelling, and degranulation of the endoplasmic reticulum. Our results suggest that nicotine causes HCs' degeneration and loss and may have implications for smoking-related hearing loss.

Puerarin alleviates the ototoxicity of gentamicin by inhibiting the mitochondria­dependent apoptosis pathway.

Gentamicin (GM) is a commonly used antibiotic, and ototoxicity is one of its side effects. Puerarin (PU) is an isoflavone in kudzu roots that exerts a number of pharmacological effects, including antioxidative and free radical scavenging effects. The present study investigated whether PU could protect against GM­induced ototoxicity in C57BL/6J mice and House Ear Institute­Organ of Corti 1 (HEI­OC1) cells. C57BL/6J mice and HEI­OC1 cells were used to establish models of GM­induced ototoxicity in this study. Auditory brainstem responses were measured to assess hearing thresholds, and microscopy was used to observe the morphology of cochlear hair cells after fluorescent staining. Cell viability was examined with Cell Counting Kit­8 assays. To evaluate cell apoptosis and reactive oxygen species (ROS) production, TUNEL assays, reverse transcription­quantitative PCR, DCFH­DA staining, JC­1 staining and western blotting were performed. PU protected against GM­induced hearing damage in C57BL/6J mice. PU ameliorated the morphological changes of mouse cochlear hair cells and reduced the apoptosis rate of HEI­OC1 cells after GM­mediated damage. GM­induced ototoxicity may be closely related to the upregulation of p53 expression and the activation of endogenous mitochondrial apoptosis pathways, and PU could protect cochlear hair cells from GM­mediated damage by reducing the production of ROS and inhibiting the mitochondria­dependent apoptosis pathway.

Downregulation of Cav3.1 T-type Calcium Channel Expression in Age-related Hearing Loss Model.

OBJECTIVE: Age-related hearing loss (AHL), characterized by degeneration of cochlea structures, is the most common sensory disorder among the elderly worldwide. The calcium channel is considered to contribute to normal hearing. However, the role of the T-type voltage-activated calcium channel, Cav3.1, remains unclear in AHL. Here, we investigate the age-related change of Cav3.1 expression in the cochlea and D-gal-induced senescent HEI-OC1 cells. METHODS: Cochleae from C57BL/6 mice at 2 months and 12 months of age were assessed. Senescence in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells was induced by D-gal treatment. The immunofluorescence technique was employed to investigate the distribution of Cav3.1 in vivo and in vitro. Quantitative assessment was achieved by Western blotting and real-time PCR. RESULTS: In comparison with 2-month-old animals, 12-month old C57BL/6 mice exhibited great loss of hair cells and elevated auditory brainstem threshold. The Cav3.1 was located in hair cells, spiral ganglion cells, lateral walls, and the expression of Cav3.1 protein and mRNA decreased in the aged cochleae. D-gal-induced senescence assay confirmed the down-regulation of Cav3.1 expression in senescent HEI-OC1 cells. CONCLUSION: Our results show that age-related down-regulated expression of Cav3.1 in the cochleae is associated with AHL and may contribute to the pathogenesis of AHL.

Endonasal Transpterygoid Surgery for Adult Mature Teratoma in the Sphenoid Bone.

ABSTRACT: Teratomas are germline tumors commonly composed of multiple cell types derived from embryonic germ cell layers. Teratomas in head and neck region are exceptionally rare and present during the neonatal and infantile period. We describe a male adult with a mature teratoma originating from sphenoid body. A 24-year-old male patient presented with left-sided intermitted headache and facial numbness. Radiographic imaging showed a 3 cm × 2.3 cm mass with heterogeneous density in the sphenoid region. The endoscopic sphenoid sinus opening surgery was performed through endonasal transpterygoid approach. The final pathologic diagnosis was confirmed as mature teratoma. The patient has been followed-up for 18 months without recurrence. We suggest endonasal transpterygoid approach could be an effective and safe treatment for patients with mature teratoma in the sphenoid bone.

Serum level and clinical significance of vitamin E in children with allergic rhinitis.

BACKGROUND: Allergic rhinitis (AR) is one of the most prevalent allergic diseases in children. This study aimed to investigate the association between serum concentrations of vitamin E and AR to determine if the vitamin E level is correlated with the occurrence and severity of AR. METHODS: A total of 113 children were enrolled in this cross-sectional study. Sixty-five children in the outpatient group were diagnosed with AR, and 48 healthy children were recruited as controls. All subjects underwent serum vitamin E (adjusted for total cholesterol and triglycerides) measurements. Serum to total IgE (tIgE), the five most common allergen-specific IgE (sIgE) levels and skin prick test (SPT) were measured in children with AR. The severity of AR was assessed with the nasal symptoms score, and the situation of exposure to passive smoking were inquired. RESULTS: Serum vitamin E levels were significantly lower in the AR group than in the normal children (P < 0.001). A significant negative correlation was observed between serum vitamin E levels and sIgE as well as the SPT grade. Serum vitamin E levels were also inversely related to the nasal symptoms score; however, statistical significance was not found. CONCLUSIONS: A significantly lower vitamin E level was found in children with AR. Lower serum vitamin E levels may have correlation with the occurrence of AR in children. However, serum vitamin E levels were not statistically correlated with the severity of AR.

lncRNA SOX2-OT regulates laryngeal cancer cell proliferation, migration and invasion and induces apoptosis by suppressing miR-654.

Laryngeal carcinoma is the most common type of malignant tumor in the head and neck. Long non-coding RNAs (lncRNAs) serve crucial roles in numerous biological processes. The present study aimed to investigate the role of lncRNA SOX2-OT in laryngeal cancer and to reveal the underlying mechanisms. Reverse transcription-quantitative PCR assays were used to measure the expression levels of SOX2-OT in the laryngeal cell lines. Furthermore, cell proliferation, apoptosis, migration and invasion were assessed by CCK-8, flow cytometry, wound healing and Transwell assays, respectively. Western blot assay was performed to detect the protein expressions. In addition, a dual-luciferase reporter assay was performed to confirm the direct interaction between SOX2-OT and microRNA (miR)-654. The data demonstrated that SOX2-OT level were significantly increased in the laryngeal cell lines. Furthermore, SOX2-OT silencing markedly promoted apoptosis and suppressed the proliferation, migration and invasion of TU-177 cells. A dual-luciferase reporter assay revealed that miR-654 was a direct target of SOX2-OT. Moreover, downregulation of miR-654 could attenuate cell apoptosis and accelerate cell proliferation, migration and invasion in TU-177 cells. In summary, the present study reported that knockdown of SOX2-OT could suppress cell proliferation, migration and invasion, and induce apoptosis in laryngeal cancer by targeting miR-654.

[Analysis of the effect of endoscopic surgery combined with conventional surgery in Neuroblastoma].

Objective:The aim of this study is to evaluate the efficacy of endoscopic surgery and conventional surgery combined with radiotherapy in the treatment of Neuroblastoma. Method:Forty-three patients with olfactory neuroblastoma undergoing surgery combined with radiotherapy were retrospectively analyzed. The patients were divided into endoscopic surgery and conventional surgery. All patients received postoperative radiotherapy at a dose of 60-70 Gy, the 5-year survival rate and local recurrence time of the two groups were compared, and the therapeutic effects of endoscopic surgery and traditional surgery were compared. Result:Through survival analysis, the 5-year overall survival rates of the traditional surgery group and the endoscopic surgery group were 50% and 58% （P=0.560）, the local recurrence rates were 44% and 48% （P=0.288）, and the mean recurrence time was 5.6 months and 12.5 months （P=0.032）. Conclusion:There was no difference between endoscopic surgery and conventional surgery combined with radiotherapy in the treatment of Neuroblastoma, and the time of local recurrence was significantly prolonged. In early Neuroblastoma, endoscopic sinus surgery may be superior to open surgery in terms of efficacy and patient survival.

Knockdown of YAP inhibits growth in Hep-2 laryngeal cancer cells via epithelial-mesenchymal transition and the Wnt/ß-catenin pathway.

BACKGROUND: Yes-associated protein (YAP) plays a crucial role in tumour development and it is the main effector of the Hippo signalling pathway. However, the mechanism underlying YAP downregulation in laryngeal cancer is still unclear. In our previous study, we found that YAP, compared with adjacent tissues, was expressed higher in laryngeal cancer and was also closely associated with histological differentiation, TNM stage and poor prognosis. METHODS: In this study, we attempted to determine whether silenced YAP could downregulate human laryngeal carcinoma Hep-2 cells progression. YAP was downregulated in Hep-2 cells by shRNA, and the malignant ability of Hep-2 was assessed in vitro and in vivo. RESULTS: In vitro, CCK-8, colony formation and wound healing assays showed that downregulation of YAP significantly reduced the rates of proliferation, migration, and invasion in Hep-2 cells. Downregulation of YAP distinctly induced G2/M cycle arrest and increased the rate of apoptosis. Accordingly, western blot assay suggested that the expression of DKK1, vimentin and ß-catenin was significantly decreased after YAP downregulated treatment, thereby indicating that YAP mediated the EMT programme and the Wnt/ß-catenin signalling pathway in carcinoma of the larynx. Furthermore, silencing of YAP suppressed Hep-2 cell tumourigenesis and metastasis in vivo. CONCLUSION: In summary, our findings demonstrated the proliferation of YAP downregulation and the invasion of Hep-2 cells via downregulating the Wnt/ß-catenin pathway in vitro and in vivo, suggesting that YAP may provide a potential therapeutic strategy for the treatment of laryngeal cancer.

Elevated expression of yes­associated protein is associated with the malignant status and prognosis of laryngeal squamous cell carcinoma.

Previous studies have demonstrated that elevated yes­associated protein (YAP) expression is associated with tumor aggression and poor prognosis in various types of human cancer. However, the clinicopathological significance and the prognostic value of YAP in laryngeal squamous cell carcinoma (LSCC) is unknown. The aim of the present study was to identify the expression pattern and prognostic significance of YAP in patients with LSCC. YAP mRNA and protein expression levels were examined in fresh and archived LSCC samples using the reverse transcription­quantitative polymerase chain reaction, immunohistochemistry (IHC) and western blotting. The association between YAP expression levels with the malignant status and prognosis of patients with LSCC was analyzed. Upregulated protein and mRNA expression levels of YAP were detected in LSCC tissues compared with paired healthy surgical margin tissues. Positive expression of YAP was identified in 84/121 (69.4%) LSCC tissues and in 4/30 (13.3%) healthy surgical margin tissues by IHC. Positive YAP protein expression was significantly associated with clinical stage, TNM classification, lymph node metastasis and differentiated degree. Patients with positive YAP expression exhibited a significantly decreased overall survival time compared with patients with negative YAP expression (P=0.0002). Multivariate analysis indicated that the level of YAP expression was an independent prognostic factor for poor survival in patients with LSCC (P=0.012). In conclusion, the expression level of YAP was significantly increased in LSCC and associated with the malignant status of LSCC. Therefore, YAP may represent a novel biomarker for predicting the prognosis of patients with LSCC.

Knockdown of ST6Gal-I increases cisplatin sensitivity in cervical cancer cells.

BACKGROUND: Sialyltransferase I (ST6Gal-I) is an enzyme involved in tumor metastasis that processes sialic acid precursors into their mature form, enabling them to regulate gene expression. However, the effect of ST6Gal-I on the biological behavior of cancer cells remain unclear. This study was the first to demonstrate the influence of ST6Gal-I on cisplatin sensitivity in cervical cancer cells. METHODS: Knockdown of ST6Gal-I was performed by shRNA and HeLa cells combination with cisplatin were tested. RESULTS: We showed that down-regulation of ST6Gal-I promoted cell apoptosis and inhibited proliferation and invasion in cervical cancer cells. Knockdown of ST6Gal-I by RNA interference increased the sensitivity of HeLa cells to cisplatin in vitro, and reduced tumor volume and suppressed subcutaneous tumor growth in response to cisplatin treatment in a xenograft mouse model in vivo. CONCLUSIONS: The results provide new information that ST6Gal-I plays an important role in several biological or pathological processes including drug resistance in cervical cancer and may be a potential therapeutic target to improve the response to chemotherapy in cervical cancer patients.

Downregulation of inwardly rectifying potassium channel 5.1 expression in C57BL/6J cochlear lateral wall.

Age-related hearing loss (AHL) is one of the most common sensory disorders among elderly persons. The inwardly rectifying potassium channel 5.1 (Kir5.1) plays a vital role in regulating cochlear K(+) circulation which is necessary for normal hearing. The distribution of Kir5.1 in C57BL/6J mice cochleae, and the relationship between the expression of Kir5.1 and the etiology of AHL were investigated. Forty C57BL/6J mice were randomly divided into four groups at 4, 12, 24 and 52 weeks of age respectively. The location of Kir5.1 was detected by immunofluorescence technique. The mRNA and protein expression of Kir5.1 was evaluated in mice cochleae using real-time polymerase-chain reactions (RT-PCR) and Western blotting respectively. Kir5.1 was detected in the type II and IV fibrocytes of the spiral ligament in the cochlear lateral wall of C57BL/6J mice. The expression levels of Kir5.1 mRNA and protein in the cochleae of aging C57BL/6J mice were down-regulated. It was suggested that the age-related decreased expression of Kir5.1 in the lateral wall of C57BL/6J mice was associated with hearing loss. Our results indicated that Kir5.1 may play an important role in the pathogenesis of AHL.

Down-regulation of the large conductance Ca(2+)-activated K(+) channel expression in C57BL/6J cochlea.

CONCLUSION: The large conductance Ca(2+)-activated K(+ )channels (BK) expression is decreased in the cochleae of age-related hearing loss (AHL) mice. BK channel may be associated with AHL. OBJECTIVE: AHL is the most common among elderly persons. BK channels act as sensors for membrane voltage and intracellular Ca(2+ )and are essential for hearing. To investigate the distribution of BK channel in the cochleae of C57BL/6J mice, and the relationship between the expression of BK channel and the etiology of AHL. METHODS: BK expression was studied in the cochleae of C57BL/6J mice at various ages (4, 12, 26, 52 weeks). The expressions of BK at the protein and mRNA levels were detected by immunofluorescence technique, western blot and quantitative real time PCR. RESULTS: In comparison with 4-week-old mice, BK expressions in the cochleae at 12, 26 and 52 weeks of age were significantly and gradually decreased at both the protein and the mRNA levels. The immunofluorescence technique showed the BK channel was located in the hair cells and cells of the spiral ganglion, spiral ligament and stria vascularis and its expression also decreased with aging.

17-DMAG induces Hsp70 and protects the auditory hair cells from kanamycin ototoxicity in vitro.

Heat shock protein 70 (Hsp70) has been known to be able to play a protective role in the cochlea. The aim of this study was to investigate whether geldanamycin hydrosoluble derivative 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG) has the ability to induce Hsp70 up-regulation to protect hair cells from kanamycin-induced ototoxicity in vitro. The organ of Corti (OC) explants were isolated from mice at postnatal day 3-5. Then, the explants were exposed to kanamycin with or without pre-incubation with 17-DMAG. The expression of Hsp70 was assessed by reverse transcription-quantitative polymerase chain reaction, ELISA, and immunofluorescent staining. The surviving hair cells were examined by phalloidin labeling and were counted. We found that Hsp70 expression in the explants after pre-incubation with 17-DMAG was significantly increased at both mRNA and protein levels. Immunofluorescent staining showed that Hsp70 was mainly located in the auditory hair cells. Compared with kanamycin group, the loss of hair cells was inhibited significantly in 17-DMAG+kanamycin group. Our study demonstrated that 17-DMAG induces Hsp70 in the hair cells, and has a significant protective effect against kanamycin ototoxicity in vitro. 17-DMAG has the possibility to be a safe and effective anti-ototoxic drug.

Age-related change in the expression of NKCC1 in the cochlear lateral wall of C57BL/6J mice.

CONCLUSIONS: Na(+)-K(+)-2Cl(-) co-transporter isoform 1 (NKCC1) mRNA and protein decrease with increasing age in the cochlear lateral wall of C57BL/6J (C57) mice. The down-regulation of NKCC1 may influence the K(+) transport efficiency and the homeostasis of ion transport cells, and cause the irreversible damage of cochlear cells in old C57 mice. Our results indicate that NKCC1 may play an important role in the pathogenesis of age-related hearing loss (AHL). OBJECTIVES: The aim of the present study was to investigate the relationship between the functional expression of NKCC1 transporter and the etiology of AHL. METHODS: C57 mice were used and randomly divided into four groups according to age (4 weeks, 14 weeks, 26 weeks, and 52 weeks). Immunofluorescence technique, quantitative real-time PCR, and western blot were applied to detect the expression of NKCC1 in the cochlear lateral wall of C57 mice at the various ages. RESULTS: In all four groups, the expression of NKCC1 was observed in the stria vascularis and type II fibrocytes of the spiral ligament. Also, the expression of NKCC1 appeared to decrease with age at both the transcriptional level and the protein level.

Expression of HMGA1 and ezrin in laryngeal squamous cell carcinoma.

CONCLUSION: The overexpression of HMGA1 or Ezrin may contribute to the carcinogenesis, development, and metastasis of laryngeal squamous cell carcinoma (LSCC). OBJECTIVE: To investigate the expression of HMGA1 and Ezrin in LSCC and analyze their clinical significance. METHODS: The expression of HMGA1 and Ezrin was analyzed by immunohistochemistry (IHC) in 50 cases of LSCC. Thirty cases of laryngeal polyp and 30 cases of atypical hyperplasia of larynx were studied as controls. The expression of HMGA1 and Ezrin was analyzed by real-time PCR and by Western blot in 30 cases of LSCC; samples from adjacent normal epithelial tissues in 30 cases were studied as controls. RESULTS: (1) IHC revealed that the positive rate of HMGA1 protein was 68.0% (34/50), 53.3% (16/30), and 13. 3% (4/30) in LSCC, atypical hyperplasia of larynx, and laryngeal polyp (p < 0.05), and the positive rate of Ezrin protein was 64.0% (32/50), 50.0% (15/30), and 23.3% (7/30) (p < 0.01), respectively. (2) Real-time PCR demonstrated that the mean relative mRNA expression levels of HMGA1 in LSCC and in normal tissues were 2.41 ± 0.40 and 1.05 ± 0.18, respectively (p < 0.01). The mRNA levels of Ezrin in LSCC and in normal tissues were 1.79 ± 0.27 and 1.04 ± 0.22, respectively (p < 0.05). (3) Western blotting revealed that the mean relative protein expression levels of HMGA1 in LSCC and in normal tissues were 1.73 ± 0.60 and 0.35 ± 0.17, respectively (p < 0.01). The protein levels of Ezrin in LSCC and in normal tissues were 1.82 ± 0.77 and 0.42 ± 0.20, respectively (p < 0.01).