| Objective:The carcinoma of larynx is one of the malignant neoplasms in otolaryngology and head neck surgery. Its incidence increased significantly in recent years. The occurrence, development and metastasis of laryngeal cancer is a process by multi-factor, multi-stage and multi-gene is involved, meanwhile by the activation of oncogenes, deactivation of tumor suppressor genes and apoptogenes uncontrolled interactions. This study through the detection CTHRC1-mRNA and protein in the throat squamous cell carcinoma (LSCC) tissue of expression, analysis of the CTHRC1expression and the clinical and pathological factors and the relationship between clinical significance. A preliminary study on the its significance in the development of laryngeal cancer, provide theory basis for the early diagnosis of laryngeal cancer and gene therapy.Methods:experimental subject:Materials from the second hospital of tianjin medical university in February2008to October2013of otolaryngology-head and neck surgery hospitalized patients with laryngeal cancer surgery, selection of laryngeal squamous carcinoma specimens of62cases were confirmed by pathology of squamous cell carcinoma, Use the international association of clinical stage (UICC) TNM classification staging standard published in2002phase Ⅰ, Ⅱ,42cases of20patients with stage Ⅲ glottis in33cases, glottis29cases. Of the glottis type Ⅰ, Ⅱ throat throat part resection, phase Ⅲ line complete excision throat or complete cut method, type Ⅰ, Ⅱ line above the glottis throat, for partial nephrectomy or horizontal partial laryngectomy phase Ⅲ throat subtotal resection or full resection,15cases obtained the glottis and the glottis cancer near normal laryngeal mucosa as control take edge distance carcinoma (5mm), confirmed by pathology in inflammation or normal mucosa, The organization of laryngeal squamous carcinoma specimens of paraffin.In order to verify the results of immunohistochemical, we collected20cases of the glottis and the glottis laryngeal squamous carcinoma tissue and adjacent5mm above the normal epithelial tissue (confirmed by pathology, no tumor tissue infiltration) of frozen specimen extraction mRNA (-80℃).18cases of male, female in2cases, high-differentiation in17cases,3cases of low differentiation,12cases Ⅰ, Ⅱ,8cases Ⅲ,5cases with lymph node metastasis, no lymph node metastasis15cases, all patients has not been preoperative radiation therapy and chemical therapy, clinical data integrity.1.Immunohistochemistry(S-P two-step methods)was employed tomeasure the expression of CTHRC1in62specimens of laryngeal squamous cell carcinoma tissue and15specimens of adjacent tissue, And detection of CTHRC1expression differences in whether there is a lymph node metastases,and Statistical analysis of their correlation with the clinical pathological factors.2.RT-PCR and Western-blotting were employed to measure the expression of CTHRC1mRNA and protein in20frozen specimens of laryngeal squamous cell carcinoma tissue and20specimens of adjacent tissue. Statistical analysis and validate their relevance to clinical pathological factors.3.Statistics analysis:SPSS for19.0software analyzes the data. The Pearson’s Chi-Square test analysis CTHRC1of the expression and throat squamous cell carcinoma patients with clinical pathological factors, P<0.05is considered statistically significant, P<0.01indicating very significant difference.Results:1.Expression of CTHRC1and relation with clinical parameter1.1Immunohistochemistry:was employed to measure the expression of CTHRC1in62specimens of laryngeal squamous cell carcinoma tissue and15specimens of adjacent tissue. the positive expression rate in Cytoplasm respectively were79.03%(49/62)and13.33%(2/15), The former is obviously higher than the latter, the two have very significant difference statistically significance (P<0.01), CTHRC1in laryngeal squamous carcinoma tissue protein expression is significantly higher than normal laryngeal mucosa tissue adjacent to carcinoma. CTHRC1express in laryngeal squamous cell carcinoma of the presence or not lymph node metastases:the CTHRC1positive expression rate in lymph node metastases was92%(23/25), the positive expression rate in no lymph node metastasis was70.27%(26/37),The positive expression rate of lymph node metastasis group is significantly higher than without lymph node metastasis (P<0.05), Protein Expression of CTHRC1was correlated with lymph node metastasis,clinical stage(P<0.05), but that wasn’t correlated with pathological grading of tumor,patients’clinical classification and age (P>0.05).1.2RT-PCRmethod:RT-PCR method for detection of20cases of laryngeal squamous carcinoma tissues and20cases of normal laryngeal mucosa tissue adjacent to carcinoma of CTHRC1mRNA expression of mRNA of laryngeal squamous carcinoma tissues of the average grey value was0.93+/-0.15, and paired normal laryngeal tissue CTHRC1mRNA in the average grey value of0.29+/-0.23, CTHRC1mRNA expression in laryngeal squamous carcinoma tissue is significantly higher than normal laryngeal mucosa tissue adjacent to carcinoma, both have very significant difference statistically significance (P<0.01)(FIG.1,2); In laryngeal squamous carcinoma CTHRC1mRNA expression and lymph node metastasis (P0.05) and clinical stage (P<0.05), and has nothing to do with tumor differentiation, sex and age groups (P>0.05).Conclusions:1.In laryngeal squamous carcinoma tissues, CTHRC1has high expression, prompt its may be associated with the incidence of laryngeal squamous carcinoma.2.CTHRC1in laryngeal squamous carcinoma tissue is high expression and associated with lymph node metastasis, clinical stage, but with tumor pathology classification, gender and age group has nothing to do, May be associated with tumor invasion and metastasis.3.CTHRC1may be related to laryngeal squamous carcinoma (HCC) development, invasion and metastasis, is expected to become the prognosis of laryngeal squamous carcinoma new therapeutic targets and indicators... |
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