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Inhibition of STAT3 expression by siRNA suppresses growth and induces apoptosis in laryngeal cancer cells

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Acta Pharmacologica Sinica 2005年第3期26卷 377-383页

作者： Li-fang GAO~1 De-qi XU~2 Lian-ji WEN~3 Xing-yi ZHANG~4 Yue-ting SHAO~1 Xue-jian ZHAO~(1,5)1 Department of Pathophysiology,Basic School of Medicine,Jilin University,Changchun 130021,China 2 Laboratory of Enteric and Sexually Transmitted Diseases,Center for Biologics Evaluation and Researeh,Food and Drug Administration,Bethesda MD,20892,USA 3 Departments of ENT4 Thoracic and Cardiovascular Surgery,the Second Hospital of Jilin University,Changchun 130021,China Department of Pathophysiology Basic School of Medicine Jilin University Changchun China Laboratory of Enteric and Sexually Transmitted Diseases Center for Biologics Evaluation and Research Food and Drug Administration Bethesda USA Departments of ENT Changchun China Thoracic and Cardiovascular Surgery the Second Hospital of Jilin University Changchun China

Aim:To determine the inhibitory effect of the synthetic STAT3 siRNA on theexpression of STAT3 gene in human laryngeal cancer cell lines Hep2 and to inves-tigate the effect of STAT3 siRNA on growth and apoptosis in Hep2 ***:A pair of DNA templates coding siRNA against STAT3-mRNA was synthesized toreconstruct plasmid of pSilencer1.0-U6 ***2 cells were trans-fected with RPMI-1640 media (untreated),plasmid (empty),and STAT3 siRNA,*** blot and Western blot analysis of STAT3 and pTyr-STAT3expression in Hep2 cells and Western blot analysis of Bcl-2 expression in the Hep2cell was performed 72 h after ***,flow cytometry,and AO/EBassay were used for determination of cells proliferation and apoptosis in ***:pTyr-STAT3 was markedly expressed in untreated Hep2 cells andthe vector-treated Hep2 cells,whereas pTyr-STAT3 expression was significantlyreduced in STAT3 siRNA-transfected Hep2 cells,indicating that STAT3 siRNAinhibited the activity of *** of Hep2 cells with STAT3 siRNAsignificantly inhibited STAT3 expression at both mRNA and protein level in Hep2cells and the inhibition was characterized by time-dependent ***-ment of Hep2 cells with STAT3 siRNA resulted in dose-dependent growth inhibi-tion of Hep2,this significantly increased apoptotic cell rate,and decreased Bcl-2expression level in Hep2 ***3 siRNA had an effect on induction of eitherearly or late stage ***:This study demonstrates that STAT3siRNA effectively inhibits STAT3 gene expression in Hep2 cells leading to growthsuppression and induction of apoptosis in Hep2 *** use of siRNA tech-nique may provide a novel therapeutic approach to treat laryngeal cancer andother malignant tumors expressing constitutively activated STAT3.

关键词： STAT3 RNA interference laryngeal neoplasms apoptosis

Effect of DRB on the Biological Characteristics of Human laryngeal Carcinoma Hep-2 Cell Line

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第1期27卷 104-106页

作者：王建亭龚树生付勇薛秋红陈广理刘英鹏 Department of Otolaryngology Union Hospital Tongji Medical College Huazhong University of Science and Technology

In order to study the effect of 5, 6-Dichloro-l-13-D-ribofuranosyl-benzimidazole （DRB） on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry （FCM） and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was （0.68±0.19）%, （1.95±0.12）%, （8.51±0.26）%, （11.26±0.17）% and （14.99±0.32）%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.

关键词： protein-serine-threonine kinases 5,6-Dichloro-1-β-D-ribofuranosyl- benzimidazole laryngeal neoplasms Hep-2 cell line

Effect of three tongue needles acupoints Lianquan(CV23) and Hegu(LI4) combined with swallowing training on the quality of life of laryngeal cancer patients with dysphagia after surgery

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Journal of Traditional Chinese Medicine 2022年第4期42卷 617-621页

作者： ZHU Xuewei LIU Minghui ZONG Minru CHEN Qianqian WANG Jianfeng Department of otolaryngology head and neck surgery China-Japan Union Hospital of Jilin UniversityChangchun 130033China Department of Chinese traditional Medicine China-Japan Union Hospital of Jilin UniversityChangchun 130033China Department of rehabilitation China-Japan Union Hospital of Jilin UniversityChangchun 130033China Department of Radiotherapy China-Japan Union Hospital of Jilin UniversityChangchun 130033China

OBJECTIVE:To evaluate the effect of acupuncture therapies administered in combination with swallowing training on the quality of life of laryngeal cancer patients with dysphagia after ***:Seventy-one postoperative patients with laryngeal cancer participated in this *** patients diagnosed with swallowing dysfunction by video fluoroscopic swallowing examination(VFSE)were randomly divided into experimental group(n=36)and control group(n=35).Patients in both groups were provided swallowing training and rehabilitation *** in the experimental group were additionally provided with acupuncture *** patients were evaluated using VFSE and MD Anderson dysphagia inventory(MDADI)and Quality of Life Questionnaire-core 30(QLQ-c30)score immediately after surgery and three months ***:The effective rate of 97.1%(n=35)and the complete remission rate of 36.1%(n=13)in the experimental group were higher than those in the control group of 60%(n=21)and 14.3%(n=5)(Plaryngeal cancer patients with dysphagia after surgery.

关键词： laryngeal neoplasms deglutition disorders acupuncture swallowing training quality of life

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ANALYSIS OF C-HA-RAS GENE AMPLIFICATION AND MUTATION IN laryngeal CARCINOMA

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Chinese Medical Sciences Journal 1995年第1期10卷 59-60页

作者：刘世喜林代诚洪邦泰黄光琦 Department of Otolaryngology First Hospital West China University of Medical Sciences Chengdu.

In order to study the altered molecular events during laryngeal carcinogenesis and elucidate the role of Haras oncogene amplification and mutation. we have examined their profile by polymerase chain reaction (PCR) and selective oligonucleotide hybridization. We analyzed the mutational status of codon 12 of Haras in 22 laryngeal carcinomas and 10 normal tissues. and found that 7 of 22 laryngeal carcinomas contained a Ha-ras mutation at codon 12. The frequency of mutation was 32%. None of the normal tissues revealed mutation. Moreover. no amplification was found in cancers when compared to the normal. Ourfindings indicated that the activated Ha-ras gene existed in laryngeal carcinoma. and activation of the Haras gene by mutation at codon 12 might play a key role in laryngeal carcinogenesis.

关键词： laryngeal neoplasms C-Ha-ras gene amplification gene mutation

EXTRACAPSULAR SPREAD IN IPSILATERAL NECK METASTASIS:AN IMPORTANT PROGNOSTIC FACTOR IN laryngeal CANCER

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Chinese Medical Sciences Journal 2006年第2期21卷 86-89页

作者： Bin Liu Chao Guan Wen-yue Ji Zi-min Pan Department of Otolaryngology the First Affiliated Hospital China Medical University Shenyang 110001

Objective To evaluate the impact of extracapsular spread （ECS） in ipsilateral neck metastasis on prognosis and its related factors in laryngeal cancer. Methods The study included 184 patients who underwent laryngectomy and simultaneous radical or modified radical neck dissection between January 1994 and December 1997 for laryngeal cancer. All of them had a complete 5-year follow-up. We used transparent lymph node detection and continuous slicing method on all neck dissection specimens. Kaplan-Meier model was used for survival analysis and the log-rank test was used to assess significance. Reults We found pathological neck metastases in 80 patients. Among them, 26 cases （32. 5% ） had ECS in ipsilateral neck. ECS incidence increased with advanced pathological N （pN） stages （pNl 3.7%, pN2a 25.0%, pN2b 50. 0%, and pN2c 55.6% ; P =0. 001）. ECS incidence also increased with number of positive nodes （1 positive node 8.6%, 2 positive nodes 33.3%, 3 and more positive nodes 66. 7% ; P 〈0. 001 ）. Incidences of contralateral neck metastases and ipsilateral neck recurrence in patients with ECS were higher than those in patients without ECS （46.2% vs. 24. 1%, P=0.046; 34.6% vs. 7.4%, P =0.002）. The 5-year survival rate of patients with ECS was significantly lower than that of patients without ECS （23.1% vs. 57.4%, P =0. 013）. Conclution ECS is an important prognostic factor in laryngeal cancer. Patients with ECS have a higher incidence of contralateral neck metastasis, so bilateral neck dissection should be selected.

关键词： laryngeal neoplasms lymphatic metastasis extracapsular spread prognosis

Prognostic Value of Ki67 and VE6F Expression in laryngeal Squamous Cell Carcinoma

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Chinese Journal of Clinical Oncology 2006年第1期3卷 20-24页

作者： Dawei Sun Yanjun Wang Weijia Kong Banghua Liu Department of Otolaryngology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

OBJECTIVE To investigate the prognostic value of measuring Ki67 and VEGF expression in laryngeal squamous cell carcinoma (LSCC). METHODS The expression of Ki67 and VEGF in 32 LSCC tissues was studied by immunohistochemical staining. Of these cases, 5 recurred (recurrent group), 3 cases metastasized (metastatic group), 8 cases died (deceased group) and 24 cased survived (survival group) over a 3 year period of follow-up after their operation. RESULTS The expression of Ki67 and VEGF in the deceased group was higher compared to that in the survival group (P<0.01). Overexpression of Ki67 was found in the recurrent group and in the metastatic group (P< 0.05). VEGF expression was higher in the recurrent group than in the non recurrent group (P<0.05). With Cox-regression of multivariate analysis, Ki67 (RR:3.236; P=0.001), the clinical T stage (RR: 1.382; P=0.029) and metastasis in lymph nodes (RR:0.316; P=0.033) were shown to be independent prognostic factors for survival of LSCC patients. CONCLUSION Ki67 and VEGF expression is related to the prognosis of LSCC. Overexpression of the 2 markers indicated poor prognosis of the disease, a finding which might be helpful for the treatment of laryngocar-cinoma.

关键词： laryngeal neoplasms carcinoma squamous cell Ki67 endothelial growth factors prognosis.

Effects of siRNA specific to the protein kinase CK2α on apoptosis of laryngeal carcinoma cells

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Chinese Medical Journal 2012年第9期125卷 1581-1585页

作者： WANG Jian-ting GONG Shu-sheng Department of Otorhinolaryngology Head and Neck Surgery Beijing Chaoyang Hospital Capital Medical University Beijing 100020 China Department of Otorhinolaryngology Head and Neck Surgery Beijing Tongren Hospital Capital Medical University Beijing 100730 China

Background The relationship between apoptosis and tumors is a major focus in cancer research. RNA interference （RNAi） technology has emerged as a very potent tool to generate cellular knockdown phenotypes of a desired gene. The aim of this study was to explore the effect of siRNA specific to the protein casein kinase 2α （CK2α） on apoptosis of laryngeal carcinoma cells and to explore possible mechanisms.Methods An siRNA expression plasmid specific to CK2α, psiRNA-hH1neo-CK2α, and a non-specific siRNA expression plasmid, psiRNA-hH1neo-cont, were constructed and transfected into Hep-2 cells by a lipofectamine method. The mRNA and protein levels of CK2α in transfected cells were determined by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting analysis. Apoptosis was measured by Annexin V-fluorescein isothiocyanate （FITC）/ propidium iodide （PI） double-staining methods. The morphological changes to Hep-2 cells were observed under transmission electron microscopy （TEM）. The levels of Bcl-2 and Bax proteins were measured by Western blotting analysis.Results Levels of CK2α mRNA and protein were significantly decreased in the psiRNA-hH1neo-CK2α group compared to the other groups （P 〈0.05）. The apoptotic rate of the psiRNA-hH1neo-CK2α transfected group was significantly higher compared to that in the untransfected group and the siRNA-hH1neo-cont transfected group （25.66%±0.83%, 3.66%±0.43%, and 5.18%±0.22%）（P 〈0.05）. Compared with the untransfected group and the siRNA-hH1neo-cont transfected group, the psiRNA-hH1neo-CK2α transfected group presented with classical ultrastructural features of apoptosis, such as karyopyknosis, chromatic agglutination adjacent to the nuclear membrane, and apoptotic bodies. Compared with the other two groups, the level of Bcl-2 protein in the psiRNA-hH1neo-CK2α transfected group was decreased （0.20±0.09 vs. 0.72±0.16, 0.56±0.11, P 〈0.01）, while the Bax protein level was increased （0.81±0.17 vs. 0.26±0.12, 0.33±0.1

关键词： RNA interference small interfering RNA laryngeal neoplasms apoptosis casein kinase 2

Reconstruction of laryngeal function in subtotal laryngectomy with preservation of monoarytenoid cartilage

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Journal of Medical Colleges of PLA(China) 2002年第4期17卷 293-295页

作者：阮炎艳陈文弦 Department of Otolaryngology Tangdu Hospital Fourth Military Medical University Xi'an 710038 China

Objective: To evaluate subtotal laryngectomy with preservation of monoarytenoid cartilage to re-construct laryngeal function in the treatment of advanced laryngeal cancer. Methods: We retrospectively re-viewed 48 patients with advanced laryngeal cancer (T3 or T4 on the primary site) treated by subtotal laryngec-tomy with preservation of monoarytenoid cartilage to reconstruct laryngeal function. Eighteen of them under-went neck dissection. Ipsilateral recurrent laryngeal nerve was preserved during this surgery. The cricoid cartilage was anastomosed to hypopharynx accordingly. Results: The overall 3- and 5-year survival rates were 81.25% (39/45) and 66% 67% (32/48), respectively. All patients had good phonation and swallowing function after surgery. Decannulation rate was 93. 75%. Conclusion: Subtotal laryngectomy with preserva-tion of monoarytenoid cartilage is satisfactory for treatment of elected advanced laryngeal cancers. The effect of this surgery is more satisfactory than that of the near-total laryngectomy (Pearson''s technic).

关键词： laryngeal neoplasms laryngectomy arytenoid cartilage survival rate decannulation rate

LOSS OF HETEROZYGOSITY ON CHROMOSOME 13 IN SQUAMOUS CELL CARCINOMAS OF THE LARYNX

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Chinese Journal of Cancer Research 1998年第1期10卷 48-50页

作者：白素娟张学王筠孙开来费声重

Objective: To locate lost region of tumor suppressor gene on chromosome 13q in squamous cell carcinoma of the larynx (LSCC) and to provide clues and evidence for discovering and locating new suppressor gene. Methods: Loss of heterozygosity (LOH) on chromosome 13q was analyzed in 58 LSCC patients by microsatellite polymorphic sequences in loci D13S765 (13q13), RB120 (13q142), D13S133 (13q143) and D13S318 (13q21) on chromosome 13 by PCR. Results: There weren't any LOH on chromosome 13q in 3 cases with preinvasive LSCC. Fortyfive percentage (24/53) of the 53 invasive LSCC cases showed LOH at one or more loci on chromosome 13q region. The highest percentage of LOH on chromosome 13q was 52% (22/53) at D13S765 locus. Conclusion: The deletion region on chromosome 13q was located near by D13S765 locus which is centromeric to RB1. In this region there is suppressor gene, which is related to the genesis and development of LSCC, possibly including RB1. The inactivation of these suppressor genes may be related to the genesis and development of invasive LSCC.

关键词： laryngeal neoplasms Human chromosomes Chromosome deletion Tumor suppressor genes Polymerase chain reaction.