Study On The Prediction Of TIMP-1 And NDRG3 Related To The Prognosis Of Laryngeal Squamous Cell Carcinoma

Laryngeal squamous cell carcinoma(LSCC)is a common type of head and neck malignancy.LSCC accounts for approximately 25% of all types of head and neck cancers.In recent years,the incidence rate of LSCC has consistently increased,and there is a trend of low age.Nevertheless LSCC treatments have been developed,a satisfactory therapeutic outcome has not been achieved yet.All of these indicate that LSCC is a serious suffering to human health.The etiology of LSCC is unknown.For the influence factors,like drinking,smoking,tumor TNM staging,tumor differentiation,cervical lymph node metastasis,operation and chemotherapy,while some are controversia.It would be meaningful if we introduce modem biotechnology into LSCC biological factor to predict morbidityof LSCC,explore new anti-LSCC biotherapy,estimateprognosis of LSCC.The first part: Upregulated TIMP-1 correlates with poor prognosis of laryngeal squamous cell carcinomaObjective: The tissue inhibitor of metalloproteinase-1(TIMP-1)is an endogenous inhibitor of matrix metalloproteinases(MMPs).TIMP-1 regulates the activity of MMPs,which are enzymes involved in the degradation of the extracellular matrix(ECM);hence,TIMP-1 may be involved in cancer invasion and metastasis,and potential biomarker of various types of human cancers.Several studies have also reported that TIMP-1 is highly expressed in LSCC,as well as tumor growth and invasion.However,the association between TIMP-1 expression and the clinical features of laryngeal squamous cell carcinoma(LSCC)is barely investigated.Materials and Methods: In this study,a total of 109 paraffin-embedded LSCC tissue samples and 28 control samples were collected from the archives of the Department of Pathology,the Affiliated Hospital of Nantong University.Among the included 109 samples,107 were male,and 2 were female.The patients aged 42-87 years.There were 51 patients with well-differentiated LSCC,53 patients with moderate-differentiated LSCC,5 patients with poor-differentiated LSCC.Totally 13 patients were in the stage I,51 in the stage II,40 in the stage III,and 5 in the stage IV.19 cases with positive lymph node metastasis and negative metastasis in 90 cases.There were 37 patients who died during the follow-up period,6 patients with local recurrence,1 patients with liver metastases,and 1 patients with pulmonary metastases.Totally 74 cases of LSCC with partial laryngectomy,35 cases of LSCC with total laryngectomy.There were 77 cases of LSCC treated with radiotherapy after surgery,and 32 patients were not.12 samples of fresh LSCC tissues and matched tumor-adjacent tissues were collected from the Department of Otolaryngology-Head and Neck Surgery,Yiji Shan Hospital of Wannan Medical College.Fresh tissues were taken from the operation,immediately placed in liquid nitrogen,and then stored in a cryogenic freezer.In this present study,one-step quantitative reverse transcription-polymerase chain reaction(q RT-PCR)and western blotting tests with 12 fresh-frozen LSCC samples and matched tumor-adjacent tissues,and immunohistochemistry(IHC)analysis in 109 LSCC cases and 28 control samples were performed to investigate the TIMP-1m RNA and protein expression.Results:Study showed that the TIMP-1 m RNA and protein expression levels were significantly higher in LSCC than in the corresponding non-cancerous tissues(p<0.05).TIMP-1 protein expression in LSCC was associated with tumor differentiation(p=0.012)and overall survival(p=0.043).Kaplan-Meier method and Cox multifactor analysis suggested that positive lymph node metastasis(p=0.029)and high TIMP-1 protein expression(p=0.008)were significantly associated with the poor survival of patients with LSCC.Conclusion: High TIMP-1 expression may serve as an independent prognostic marker of LSCC.TIMP-1 overexpressionwas correlated with an aggressive malignant phenotype of LSCC.TIMP-1 may be identified as a prognostic marker of LSCC.The second part : High expression of NDRG3 associates with positive lymph node metastasis and unfavourable overall survival in laryngeal squamous cell carcinoma.Objective: The human N-myc downstream-regulated gene family(NDRG)has recently been classified as a novel type of Myc-repressed gene,named by the inhibition of Myc family cell transformation and transcription,and is involved in numerous cellular activities,including cell proliferation,apoptosis,differentiation and pathogenesis.NDRG3 belongs to the NDRG family,is believed to play important roles in human cancer.Materials and Methods: In this study,the clinical parameters of 109 LSCC cases and 28 cases of corresponding non-cancerous tissues were identical with the first part,10 samples of fresh LSCC tissues and corresponding non-cancerous tissues were collected from the Department of Otolaryngology-Head and Neck Surgery,Yiji Shan Hospital of Wannan Medical College.In this present study,q RT-PCR and western blotting tests with 10 fresh-frozen LSCC tissues and corresponding non-cancerous tissues and IHC analysis in 109 LSCC cases and 28 cases of corresponding non-cancerous tissues were performed to investigate the NDRG3 m RNA and protein expression.Results:Study demonstrated that NDRG3 m RNA and protein expression levels were statistically higher in LSCC tissues than that in non-cancerous tissues(all p<0.05).IHC data showed that the NDRG3 protein expression was remarkably correlated with lymph node metastasis(p=0.043).Univariate and multivariate survival analysis and Kaplan-Meier method implied that high NDRG3 expression(p=0.004),lymph node metastasis(p=0.044)and TNM stage(p=0.020)were independently associated with the unfavourable overall survival of patients with LSCC.Conclusion: NDRG3 overexpression was correlated with an aggressive malignant phenotype of LSCC.NDRG3 may be identified as a novel biomarker predicting the prognosis of LSCC.