Chen-Yu Chien

Calreticulin, an endoplasmic reticulum-resident protein, is highly expressed and essential for cell proliferation and migration in oral squamous cell carcinoma

OBJECTIVES Oral squamous cell carcinoma (OSCC) has emerged as one of the major malignant tumors of the head and neck cancers. However, the molecular mechanism behind tumorigenesis of OSCC is not fully understood. The aim of this study was to investigate the role of calreticulin (CRT), an endoplasmic reticulum-resident protein, in OSCC cells. MATERIALS AND METHODS Sixteen paired samples of tumor and non-cancerous matched tissue (NCMT), six OSCC cell lines and normal human oral keratinocytes (NHOKs), and oral tissue microarray were used to reveal the expression of CRT by Western blotting and immunohistochemistry. Later, shRNA-mediated stable knockdown of CRT in OSCC cells was generated. The knockdown cell line was used to analyze cell proliferation, colony formation, anchorage-independent growth and cell migration in vitro. RESULTS CRT was differentially expressed in fresh tumor samples and six OSCC cell lines but not adjacent NCMTs and NHOKs. In oral tissue microarray, we showed that there was positive CRT staining in the vast majority of tumor cases (99/103), in sharp contrast to that in NCMT cases (29/92) (p<0.001). Stable knockdown of CRT in oral cancer cells resulted in significantly reduced growth rate, colony-forming capacity and anchorage-independent growth. This may be attributed to the induction of G0/G1 cell cycle arrest when CRT was depleted in the cells. Both horizontal and vertical movements of the CRT-knockdown stable line were markedly impaired. The phosphorylation levels of focal adhesion kinase (FAK), paxillin and ERK1/2 and the activity of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) were decreased in the CRT-knockdown cells. These results suggest that CRT can regulate oral cancer cell migration through activation of the FAK signaling pathway accompanied with proteolytic degradation of the extracellular matrix (ECM) by MMP-2 and MMP-9. CONCLUSION Together, this study has defined a novel biological role for CRT in oral cancer. CRT is a potential biomarker and may contribute to the malignant phenotypes of OSCC cells.

Serum 25-hydroxyvitamin D levels are lower in chronic rhinosinusitis with nasal polyposis and are correlated with disease severity in Taiwanese patients.

Background Vitamin D deficiency is reported to be associated with increased incidence of allergic airway diseases and is correlated with the severity of asthma. This study was designed to determine if serum Vitamin D level is lower in chronic rhinosinusitis with nasal polyposis (CRSwNP) patients and if low serum Vitamin D level is correlated with the severity of CRSwNP. Methods New CRSwNP patients undergoing elective endoscopic sinus surgery were recruited. Patients with malignancies or asthma were excluded. Twenty chronic rhinosinusitis without nasal polyposis (CRSsNP) patients were used as control. Demographic characteristic information was collected. The severity of CRSwNP was assessed with the Lund-Mackay (LM) score and polyp grading system. Vitamin D status was assessed by measuring circulating 25-hydroxyvitamin D (25OHD) by using commercial chemiluminescence immunoassay. Data were stratified by factors known to affect serum 25OHD, including sex, race, and body mass index. Results Serum 25OHD levels (ng/mL ± SD) were significantly lower in patients with CRSwNP (21.4 ± 5.7) than in those with CRSsNP (28.8 ± 6.2; p < 0.001). The incidences of vitamin D deficiency (<20 ng/mL) in CRSwNP and CRSsNP patients were 45.5 and 6.3%; however, the incidences of vitamin D insufficiency (20–30 ng/mL) in these patients were 50.0 and 62.5%, respectively. A significantly negative relationship was found between serum 25OHD level and polyp grade (r = −0.63; p = 0.001), indicating lower serum 25OHD was associated with higher polyp grade. Serum 25OHD was inversely related to both LM score and total IgE level as well; however, statistical significance was not found. Conclusion A significantly lower vitamin D level was found in a group of Taiwanese CRSwNP patients, which revealed an association with greater nasal polyp size. Serum vitamin D levels could be added to the routine workup of patients suffering from CRS and these data could be used to potentially help determine the disease severity.

Association of Polymorphisms of Heat Shock Protein 70 with Susceptibility to Noise-Induced Hearing Loss in the Taiwanese Population

Noise-induced hearing loss (NIHL) is the major cause of adult sensorineural hearing loss. It is a complex disease caused by the interaction of environmental and genetic factors. Previous studies found that heat shock proteins (HSPs) were associated with the development of NIHL. Specifically, polymorphisms in the heat shock protein 70 (HSP70) gene family are associated with a susceptibility to NIHL. In this study, three single nucleotide polymorphisms (SNPs) of the HSP70 family (SNP1: rs2075800; SNP2: rs1043618; SNP3: rs2763979) were genotyped in 349 noise-exposed Taiwanese workers. The subjects were categorized into noise-susceptible (NS; n = 27) and general susceptibility (GS; n = 322) groups by the change of a 4K-weighted audiometric average in an interval of 5 years. The G/C genotype of SNP2 was found to be associated with NIHL susceptibility (adjusted OR = 2.634; 95% CI = 1.096–6.328). No significant association was found for SNP1 and SNP3 with NIHL susceptibility. Analysis of haplotypes composed of these three SNPs revealed a significant association between NIHL susceptibility and haplotype CCC (OR = 2.197; 95% CI = 1.110–4.370). In conclusion, the genetic polymorphisms in the HSP70 genes seem to be associated with the individual’s susceptibility to NIHL in the Taiwanese population. These findings could be used as a reference in the understanding and prevention of NIHL.

Heat Shock Protein 70 Gene Polymorphisms in Sudden Sensorineural Hearing Loss

Background: Heat shock proteins protect cells and tissues against different types of damage. Previous studies have revealed that the serum level of heat shock protein 70 (HSP70) increases in sudden sensorineural hearing loss (SSNHL) patients. We hypothesized that genetic variants of the HSP70 gene are associated with susceptibility to SSNHL. Methods: We conducted a case-control study with 160 SSNHL cases and 178 controls. Three tagging single nucleotide polymorphisms (SNPs) were selected. The genotypes were determined using TaqMan technology. Hardy-Weinberg equilibrium was tested for each SNP, and genetic effects were evaluated according to three inheritance modes. A haplotype analysis was also performed. Results: All three SNPs were in Hardy-Weinberg equilibrium. The CT genotype of rs2075800 exhibited an adjusted odds ratio of 0.59 (95% confidence interval 0.37–0.94; p = 0.027). The T allele of SNP rs2075800 was associated with SSNHL under the dominant model (p = 0.019; odds ratio 0.59). Haplotype analysis of the three SNPs demonstrated that the haplotype TGC (rs2075800/rs1043618/rs2763979) was statistically significant (p = 0.0137). Conclusions: These results suggest that HSP70 gene polymorphisms influence the susceptibility to the development of SSNHL in the Taiwanese population.

Metabolic Syndrome Increases the Risk of Sudden Sensorineural Hearing Loss in Taiwan A Case-Control Study

Objective Sudden sensorineural hearing loss has been reported to be associated with diabetes mellitus, hypertension, and hyperlipidemia in previous studies. The aim of this study was to examine whether metabolic syndrome increases the risk of sudden sensorineural hearing loss in Taiwan. Study Design A case-control study. Setting Tertiary university hospital. Subjects and Methods We retrospectively investigated 181 cases of sudden sensorineural hearing loss and 181 controls from the Department of Otorhinolaryngology, Kaohsiung Medical University Hospital, in southern Taiwan from 2010 to 2012, comparing their clinical variables. We analyzed the relationship between metabolic syndrome and sudden sensorineural hearing loss. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III with Asian modifications. The demographic and clinical characteristics, audiometry results, and outcome were reviewed. Results Subjects with metabolic syndrome had a 3.54-fold increased risk (95% confidence interval [CI] = 2.00-6.43, P < .01) of having sudden sensorineural hearing loss compared with those without metabolic syndrome, after adjusting for age, sex, smoking, diabetes mellitus, hypertension, and hyperlipidemia. With increases in the number of metabolic syndrome components, the risk of sudden sensorineural hearing loss increased (P for trend <.01). Vertigo was associated with a poor outcome (P = .02; 95% CI = 1.13~5.13, adjusted odds ratio = 2.39). The hearing loss pattern may influence the outcome of sudden sensorineural hearing loss (P < .01). Conclusion These results suggest that metabolic syndrome is an independent risk factor for sudden sensorineural hearing loss in Taiwan. Vertigo and total hearing loss were indicators of a poor outcome in sudden sensorineural hearing loss.

Chronic obstructive pulmonary disease predicts chronic rhinosinusitis without nasal polyps: A population-based study.

Background Few studies have investigated the relationship between chronic obstructive pulmonary disease (COPD) and chronic rhinosinusitis without nasal polyps (CRSsNP) outcomes. The aim of this study was to investigate the association between COPD and the risk of CRSsNP in a large national sample. Methods Patients 15 years or older with a new primary diagnosis of COPD (International Classification of Diseases, Ninth edition [ICD-9], 491, 492, 494, and 496) between 2000 and 2007 were identified from the National Health Insurance Research Database of Taiwan. The patients were compared with sex-, age-, residence-, and insurance premium–matched controls, and both groups were followed up until the end of 2008 for instances of CRSsNP, defined as ICD-9 codes CRS (473, 473.0, 473.1, 473.2, 473.3, 473.8, and 473.9), excluding NP (471, 471.0, 471.1, 471.8, and 471.9). Competing risk-adjusted Cox regression analyses were applied after adjusting for sex, age, residence, insurance premium, steroid use (topical or systemic), hyperlipidemia, diabetes, hypertension, coronary artery disease, hospital admission days, and mortality. Results We included 34,029 cases and 34,029 matched controls in this study. Among the 68,058 subjects, 569 developed CRSsNP during a mean (standard deviation [SD]) follow-up period of 5.0 years (SD 2.2 years). COPD was an independent predictor of CRSsNP in the fully adjusted model (hazard ratio = 3.24; 95% CI = 2.65-3.96; p < 0.01). Conclusion COPD was associated with an increased risk of CRSsNP in this study population, independent of a number of potential confounding factors.

Vitamin D decreases the secretion of eotaxin and RANTES in nasal polyp fibroblasts derived from Taiwanese patients with chronic rhinosinusitis with nasal polyps

Eosinophils are important inflammatory cells involved in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). Vitamin D and its derivatives, in addition to their classic role as regulators of electrolytes homeostasis, have modulatory effects in immunological and inflammatory responses. Such properties suggest that vitamin D might also play a role in inflammatory airway diseases such as CRSwNP. In this study, we investigated the effect of vitamin D derivatives (calcitriol and tacalcitol) on the secretion of eotaxin and Regulated on Activation, Normal T Cell Expressed and Secreted (RANTES), the two major eosinophil chemoattractants, in fibroblasts derived from the polyps of Taiwanese CRSwNP patients. Patients diagnosed with eosinophilic CRSwNP but without malignancies or asthma and undergoing elective endoscopic sinus surgery were recruited. Three primary fibroblast cultures were established using the polyp specimens obtained from these patients. The third to eighth passages of the fibroblasts were used for in vitro studies. Nasal polyp‐derived fibroblasts were stimulated with IL‐1β (10 ng/mL) for 24 hours, followed by replacement with media alone or with calcitriol or tacalcitol (10μM) and incubation for another 24 hours. After the treatments, the levels of secreted eotaxin and RANTES were evaluated by ELISA assays. The results showed that IL‐1β could substantially stimulate the secretion of eotaxin (p < 0.01) and RANTES (p < 0.01) in nasal polyp‐derived fibroblasts. More importantly, this stimulatory effect was significantly suppressed by adding calcitriol (p ≤ 0.002 for eotaxin and p ≤ 0.008 for RANTES) or tacalcitol (p ≤ 0.009 for eotaxin and p ≤ 0.02 for RANTES). Therefore, the inhibitory effect of vitamin D derivatives on eotaxin and RANTES secretion might shed light not only on the disease mechanism, but also on the potential use of vitamin D in pharmacotherapy of Taiwanese patients with CRSwNP.

Expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 in recurrent chronic rhinosinusitis with nasal polyposis

Matrix metalloproteinase (MMP) is involved in the upper airway remodeling process. We hypothesized that MMP had an additive effect on the formation of recurrent nasal polyp. We also investigated the association between the functional promoter polymorphism of MMPs and the intensity of labeling index. Expressions of MMP‐2 and MMP‐9 were assessed via immunohistochemical staining and compared between different groups, including recurrent nasal polyps, nonrecurrent nasal polyps, and control nasal mucosa. Two promoter functional single‐nucleotide polymorphisms (rs3918242 for MMP‐9 and rs243865 for MMP‐2) were selected to correlate with staining intensity. Expression of MMP‐9 was significantly enhanced in gland for recurrent nasal polyp (p = 0.016) and nonrecurrent nasal polyp (p = 0.005) compared to the control. MMP‐2 positivity was significantly increased in surface epithelium for recurrent nasal polyp (p = 0.004) compared to the control (p = 0.061). However, there was no significant difference in MMP‐9 and MMP‐2 expressions between recurrent and nonrecurrent nasal polyps. Genetic polymorphism of MMP‐2 and MMP‐9 functional promoters was not associated with the intensity of labeling index. These results suggested that up‐regulation of MMP‐9 in gland and MMP‐2 in surface epithelium was characteristic of both recurrent and nonrecurrent nasal polyps. Pathogenesis of recurrent nasal polyps may involve a mechanism other than MMP.

A Comparative Study of Endoscopic and Microscopic Approach Type 1 Tympanoplasty for Simple Chronic Otitis Media.

OBJECTIVE Tympanoplasty is a common surgery for chronic otitis media. We analyzed the results of endoscopic and microscopic approaches for type 1 tympanoplasty in patients with simple chronic otitis media. MATERIALS AND METHODS We evaluated the records of 95 patients (100 ears) who underwent type 1 tympanoplasty from 2011 to 2014. Group 1 underwent tympanoplasty with a microscopic approach (50 ears), and Group 2 underwent tympanoplasty with an endoscopic approach (50 ears). The epidemiological profiles and postoperative results, including hearing gain, duration of surgery, perioperative nausea or vomiting, and graft success rate, were reviewed. RESULTS The epidemiological profiles and preoperative hearing status were similar in both groups. Postoperatively, both groups had equal improvements in hearing and air-bone gap as well as equal perforation rates. However, the endoscopic group suffered less perioperative nausea or vomiting and had a shorter operative time. CONCLUSION The endoscopic approach for tympanoplasty offers superior visualization and shorter operative time than conventional surgery, in addition to equal hearing outcomes and perforation rates. Furthermore, observations of fewer tissue injuries, better cosmetic outcomes, and lesser perioperative nausea and vomiting suggest that the endoscopic approach is a better choice for surgery.

Asthma associated with chronic rhinosinusitis: a population-based study.

Few studies have investigated the relationship between asthma and chronic rhinosinusitis (CRS). The present study investigated the association between asthma and the risk of CRS in a large national sample.