The Expression Of Twist,e-cadherin And N-cadherin In Laryngeal Squamous Cell Carcinoma And The Correlation

Objective:The laryngeal squamous cell carcinoma(LSCC)is one of the malignant neoplasm in otolaryngology and head and neck surgery,with a increasing tendency recently.As same as other maligament tumors,the LSCC is concerned with related genetic expression which caused by various factors. The Twist gene protein is a highly conserved transcription factor that belongs to the family of basic helix-loop-helix proteins,and it plays an important role in the embryogenesis.It has been shown that Twist is a key regulator of epithelial-mesenchymal transition(EMT) and responsible for the invasion and metastasis of tumor.The aim is to investigate the expression of Twist,E-cadherin and N-cadherin gene in laryngeal squamous cell carcinoma,to analyze their role in the tumorigenesis, progression, invasion and metastasis as well as their correlation.Methods:1 Immunohistochemistry was employed to measure the expression of Twist, E-cadherin and N-cadherin in 49 specimens of LSCC tissue and 20 specimens of adjacent tissue.2 RT-PCR was employed to measure the expression of Twist,E-cadherin and N-cadherin mRNA in 49 specimens of LSCC tissue and 20 specimens of adjacent tissue.3 Statistics:SPSS 13.0 was applied to analyze the results of experiment.P≤0.05 indicating significant difference.P≤0.01 indicating very significant difference.Results:1 Expression of Twist and relation with clinical parameter1.1 Immunohistochemistry was employed to measure the expression of Twist in 49 specimens of LSCC tissue and 20 specimens of adjacent tissue.The positive expression rate of Twist protein in LSCC tissues was 61.22% (30/49),while in the specimens of adjacent tissue was 25.00%(5/20).There was very significant difference between the two groups(P<0.01).The expression of Twist gene protein in LSCC tissues was significantly higher than that in the specimens of adjacent tissues.Expression of Twist was correlated with lymph node metastasis(the expression in N+ is stronger than N0,P<0.05),clinical stage(the expression inⅢ,Ⅳis stronger thanⅠ,Ⅱ,P<0.01),but that wasn't correlated with pathological grading of tumor,patients'clinical classification and age(P>0.05).1.2 RT-PCR was employed to measure the expression of Twist mRNA in 49 specimens of LSCC tissues and 20 adjacent tissues.In LSCC tissues and adjacent tissues,the expression of Twist mRNA were (0.93±0.39)and (0.57±0.24),there is statistic significance between them,the former was stronger than the latter (P<0.01).Expression of Twist mRNA in LSCC tissues was correlated with lymph node metastasis(the expression in N+ is stronger than N0,P<0.05),clinical stage(the expression inⅢ,Ⅳis stronger thanⅠ,Ⅱ, P<0.05),but that wasn't correlated with pathological grading of tumor,patients'clinical classification and age(P>0.05).2 Expression of E-cadherin and relation with clinical parameter2.1 Immunohistochemistry was employed to measure the expression of E-cadherin in 49 specimens of LSCC tissues and 20 adjacent tissues.The positive expression rate of E-cadherin protein in LSCC tissues was 51.02% (25/49),while in the adjacent tissues was 100.00%(20/20).There was significant difference in the positive rates of E-cadherin protein between LSCC tissues and adjacent tissues (P<0.01).Expression of E-cadherin in LSCC tissues was correlated with pathological grading of tumor(the expression in G1,G2is stronger than G3,P<0.05),lymph node metastasis(the expression in N0 is stronger than N+,P<0.01),clinical stage(the expression inⅠ,Ⅱis stronger thanⅢ,Ⅳ,P<0.01),but that wasn't correlated with patients'clinical classification and age(P>0.05).2.2 RT-PCR was employed to measure the expression of E-cadherin mRNA in 49 specimens of LSCC tissues and 20 adjacent tissues.In LSCC tissues and the adjacent tissues,the expression of E-cadherin mRNA were (0.64±0.30) and (0.93±0.33), the former was significantly lower than the latter(P<0.01). Expression of E-cadherin was correlated with pathological grading of the tumor(the expression in G1,G2is stronger than G3,P<0.01),lymph node metastasis(the expression in N0 is stronger than N+,P<0.01) and clinical stage(the expression inⅠ,Ⅱis stronger thanⅢ,Ⅳ,P<0.05),but wasn't correlated with patients'clinical classification and age(P>0.05).3 Expression of N-cadherin and relation with clinical parameter3.1 Immunohistochemistry was employed to measure the expression of N-cadherin in 49 specimens of LSCC tissues and 20 adjacent tissues.The positive expression rate of N-cadherin protein in LSCC tissues was 73.47% (36/49),that in the adjacent tissues was 40.00%(8/20).There was significant difference in the positive rates of N-cadherin protein between the two groups(P<0.05).Expression of N-cadherin in LSCC tissues was correlated with lymph node metastasis(the expression in N+ is stronger than N0, P<0.01),clinical stage(the expression inⅢ,Ⅳis stronger thanⅠ,Ⅱ,P<0.01) and age(the expression in≤60 years old is stronger than >60 years old,P<0.05),but wasn't correlated with pathological grading of the tumor and patients'clinical classification(P>0.05).3.2 RT-PCR was employed to measure the expression of N-cadherin mRNA in 49 specimens of LSCC tissues and 20 adjacent tissues.In LSCC tissues and the adjacent tissues,the exprssion of N-cadherin mRNA were 0.70±0.21 and 0.36±0.11,the former was significantly higher than the latter (P<0.01).Expression of N-cadherin mRNA in LSCC tissues was correlated with lymph node metastasis(the expression in N+ is stronger than N0, P<0.01),clinical stage(the expression inⅢ,Ⅳis stronger thanⅠ,Ⅱ,P<0.01) and age(P<0.05),but wasn't correlated with pathological grading of tumor and patients'clinical classification(P>0.05).4 Their correlations in mRNA and protein:Both mRNA and protein,There were negative correlations between Twist and E-cadherin expression (r=-0.760,P<0.01;r=-0.662,P<0.01) and between N-cadherin and E-cadherin (r=-0.615,P<0.01;r=-0.558,P<0.01) expression in LSCC tissues.There was a positive correlation between Twist and N-cadherin in LSCC tissues (r=0.850, P<0.01;r=0.649, P<0.01).Conclusions:1 In the level of mRNA and protein,the higher expression of Twist and N-cadherin and the lower expression of E-cadherin in LSCC were correlated with tumorigenesis and progession of laryngeal carcinoma.2 The higher expression of Twist and N-cadherin and the lower expression of E-cadherin in LSCC were correlated with malignant biological behavior, lymph node metastasise and clinical stage in laryngeal carcima.These proteins may play an important role in the processes of invasion and metastasia of laryngeal carcima.3 There were negative correlations between Twist and E-cadherin expression and positive correlation between Twist and N-cadherin expression in LSCC tissues,Twist is related to invasion and metastasis of laryngeal carcinoma by suppressing E-cadherin expression and promot N-cadherin expression.