| Objective Squamous cell carcinoma of the head and neck is a common malignancy, advanced oral squamous cell carcinoma often violated peritumoral soft tissue, accompanied by neck or distant lymph node metastasis, the study showed recurrence and lymph node metastasis of tongue squamous cell carcinoma prognosis Key factor. Despite the comprehensive treatment based on surgery has made great progress, but the 5-year survival rate is still hovering around 50%, it is difficult to effectively improve. So advanced squamous cell carcinoma of head and neck cancer is still in the field of refractory disease to clarify its occurrence, transfer mechanism, to find new treatments and more attention. In recent years, studies have shown that Bcl-2, Twist1, EMT plays an important role in tumorigenesis, development, in particular invasion and metastasis, but Bcl-2 and Twist1 whether synergy with each other through the mechanism of EMT involved in tongue squamous cell invasion and metastasis even explore less, in the present study Yi investigate Bcl-2, Twist1 expression and colocalization in tongue squamous cell carcinoma in a relationship with tongue squamous cell carcinoma clinical and pathological features and regulation between the EMT.Methods 1.Collection of specimens: the experimental group and 82 cases with tongue squamous cell carcinoma surgical specimens, 24 cases of the control group adjacent carcinoma; by immunohistochemical staining of tongue squamous cell carcinoma surgical specimens Bcl-2, Twist1, calcineurin cad, vimentin, cadherin expression; Bcl-2, Twist1 and neural cadherin, vimentin, protein, calcium relationship between the ?2 test or Fisher exact test clinical and pathologic features of tongue squamous cell adhesion. Application of SPSS 16.0 statistical software for statistical analysis, count data using the x2 or Fisher's exact test and Spearman correlation analysis statistical row. 2.Western blotting, Bcl-2, Twist1, EMT-associated factor expression; co-precipitation method to detect Bcl-2, Twist1 mutual phase was the case through immunization; immunofluorescence method to detect Bcl-2, Twist1 of localization.Result 1. Immunohistochemistry Results: Bcl-2?Twist1 and EMT-related factors and clinical pathology correlation between its expression in squamous cell carcinoma of the tongue: Immunohistochemical staining showed that Bcl-2 expression localized in the cytoplasm and nucleus. Twist1 expression located in the nucleus, the expression of vimentin positive signals localized in the cytoplasm, the neural cadherin, cadherin positive signals were localized in the cell membrane, a few expressed in the cytoplasm. 82 cases of oral squamous cell carcinoma in 46 cases of positive expression Twist1, adjacent tissues Twist1 four cases were positive. Twist1 expression in oral squamous cell carcinoma was significantly higher than adjacent tissues(P = 0.001). Bcl-2 expression seen in next 48 cases and six cases of oral squamous cell carcinoma tissue, the expression of Bcl-2 in oral squamous cell carcinoma and adjacent carcinoma are also statistically significant difference(p = 0.005). Bcl-2, Twist1, neural cadherin, vimentin expression in poorly differentiated group and lymph node metastasis group were significantly higher than those in well-differentiated group and non-lymph node metastasis group(Bcl-2: P = 0.040 and P = 0.012; Twist1 : P = 0.013 and P = 0.003; neural cadherin: P = 0.014 and P = 0.044; for vimentin: P = 0.041 and P = 0.012), whereas cadherin expression in well-differentiated and non-lymph node metastasis group was higher than in poorly differentiated group and lymph node metastasis(P <0.05). Bcl-2, Twist1, neural cadherin, vimentin, cadherin in patients with different gender, age, tumor T stage, the expression of the difference was not statistically significant drinking, Spearman correlation analysis Bcl-2, Twist1 factors associated with EMT, Bcl-2 expression and Twist1, neural cadherin, vimentin was positively correlated with the cadherin expression on the contrary; Twist1 expression of Bcl-2, neural cadherin, vimentin expression was positively correlated with calcium adhesive protein expression was negatively correlated. 2. In vitro experimental section: Western blot tongue squamous cell carcinoma TB3.1 of Bcl-2, Twist1, EMT-associated factor expression: Bcl-2, Twist1, neural cadherin and vimentin with prolonged hypoxia significantly upregulated, and calcium cohesin significantly reduced; immunoprecipitation assay Bcl-2, Twist1 mutual phase with, the results show: normal and lack of oxygen conditions were seen in Bcl-2 antibody deposition in the cell lysate Twist1 positive, Twist1 antibody settlement reverse cell lysate verification also found Bcl-2 positive. Thus, Bcl-2, Twist1 tongue cancer in memory in direct interaction, that is, both the formation of Bcl-2 / Twist1 complex; Immunofluorescence Bcl-2, Twist1 protein expression: Bcl-2, Twist1, significantly increased and there is positioned coincident phenomenon after hypoxia.Conclusions 1.Compared with adjacent tissues, Bcl-2, twist1 was highly expressed in squamous cell carcinoma of the tongue, explained Bcl-2, twist1 high expression of the presence of tongue squamous cell. Tongue squamous cell Bcl-2, twist1 expression and histological grade, lymph node metastasis. Tip Bcl-2, twist1 abnormal expression can promote the development of oral squamous cell carcinoma. 2. EMT neural cadherin-associated protein, vimentin, Cadherin in tongue squamous cell carcinoma with lymph node metastasis group and non-metastasis group were significantly different expression, prove EMT can promote lymph node metastasis. Bcl-2, Twist1, neural cadherin, vimentin expression in poorly differentiated group and lymph node metastasis group were significantly higher than those in well-differentiated group and non-lymph node metastasis, and in well-cadherin group and non-lymph node metastasis group group was higher than that in poorly differentiated and lymph node metastasis, Bcl-2, expressing deletion twist1 may participate in tongue squamous cell carcinoma EMT process, and can affect tumor invasion by regulating EMT, transfer, and therefore detect Bcl-2, the expression level of the protein may serve as twist1 the ability to predict lymph node metastasis reference index. 3. Western blotting, Bcl-2, Twist1, EMT-associated factor expression: Bcl-2, Twist1, neural cadherin and vimentin with prolonged hypoxia significantly upregulated, and calcium significantly reduced cohesin further evidence Bcl-2, twist and EMT-associated protein correlation. Co-immunoprecipitation assay Bcl-2, Twist1 phase with each other, the results showed that: under normal and hypoxic conditions were seen in Bcl-2 antibody deposition in the cell lysate Twist1 positive, Twist1 antibody settlement reverse cell lysate same verification appears Bcl-2 positive. Thus, Bcl-2, Twist1 tongue squamous cell memory in direct interaction, that is, both to detect Bcl-2 is formed Bcl-2 / Twist1 complex, immunofluorescence, Twist1 protein expression: hypoxia after Bcl- 2, Twist1, significantly increased and there is positioned coincident phenomenon, these studies suggest, tongue squamous cell Bcl-2 may be the mechanism by certain Twist1 shuttle into the nucleus and forms a complex combination of tongue squamous cell thus contributing to the occurrence of EMT. |
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