| Objective:This research through the detection of tumor suppressor gene P16 methylation level and vascular endothelial growth factor (VEGF-C) expression in tongue squamous carcinoma tissues, research mechanism of tongue cancer lymph node metastases in both correlation and function of susceptible for tongue cancer early diagnosis and decrease the rate of lymph node metastasis provides the theory basis for improve the prognosis.Methods:Using methylation specific polymerase chain reaction technique and immunohistochemical technique,30 cases of lymph node metastasis was detected in tongue cancer group (experimental group) and 30 cases of tongue cancer without lymph node metastases group (control group), the study of tumor suppressor gene P16 methylation in the promoter region of the degree and the P16 protein and the expression of VEGF-C, combined with the clinical pathologic data (gender, age, position, histopathological differentiation of tumor invasion, with and without lymph node metastasis and other factors) were analyzed, and explores the process of tongue cancer lymph metastasis suppressor gene P16 methylation level and the correlation of expression of VEGF-C.Results:_Fluorescent methylation specific polymerase chain reaction Q-MSP part:the experimental group 30 cases of lymph node metastasis tongue cancer tissue, Q-MSP qualitative tests, using methylation primer sequence, reasonable detect p16 CpG island methylation amplification graphics, CP value between 20 to 30, methylation positive rate was 76.66%(23/30), the control group of 30 patients with no lymph node metastasis in tongue cancer tissue, using the same methylation primer sequences for testing, p16 methylation positive rate was 36.66%(11/30), the experimental group and control group in both data comparison difference to be markedly, with statistical significance (P< 0.05);At the same time the use of methylation in primer sequences, the experimental group 30 cases of tongue cancer organization gain amplification figure 63.33% (19/30), the control gain reasonable expansion graph was 26.66%(8/30), CP value between 20 to 30, the experimental group and control group in both data comparison difference to be markedly, with statistical significance (P< 0.05).Immunohistochemical experiment part:the rate of P16 protein expression was 20.00%(6/30), the control rate of P16 protein expression was 56.66%(17/30), between statistically significant (P< 0.05).By computing the correlation coefficient of phi P16 methylation and the relationship between P16 protein expression, the results of r= 0.7841, P16 gene methylation and P16 protein has strong correlation;VEGF-C positive expression rate in the experimental group was 89.38%(26/30),30 cases of lymph node metastasis in tongue cancer of VEGF (-) 4 cases. (+) in 11 cases. (++) 15 cases;VEGF-C positive expression rate in the control group was 25.93%(7/30), VEGF in 30 cases of tongue cancer (-),23 cases. (+) (3 cases), (++) 4 cases, significantly lower than that of the experimental group, so the VEGF-C positive expression rate in tongue cancer lymph node metastasis and without lymph node metastasis tissue between the difference statistically significant (P< 0.05).Different gender, age, and tumor invasion parts of tongue cancer tissues p16 gene methylation expression degree has no statistical significance (P> 0.05), the degree of histological differentiation, presence of lymphatic metastasis of tongue cancer tissue expression of p16 gene methylation degree of statistical significance (P< 0.05);Area of different gender, age, tumor invasion and differentiation degree of tongue cancer tissue VEGF-C expression rate of no statistical significance (P> 0.05), with and without lymph node metastasis of tongue cancer tissue in the expression of VEGF-C rate was statistically significant (P< 0.05).Experimental group by statistical phi correlation coefficient calculation tongue cancer lymph node metastasis tissues of P16 gene and the expression of VEGF-C correlation, the results of r= 0.5749, confirmed that P16 gene and VEGF-C have moderate correlation in tongue cancer lymph node metastasis.Conclusion:1, P16 methylation and the expression of VEGF-C and patients’gender, age, tumor invasion parts, such as clinical pathologic factors have no obvious correlation with cancer tissue differentiation degree, whether the presence of lymph node metastasis is closely related to the factors such as;2, P16 methylation has strong correlation degree and P16 protein expression, P16 methylation is one of the important inducing factors lead to tumorigenesis and development, to participate in the various stages in the process of tumor development;3, the VEGF-C expression rate of lymph node metastasis in tongue cancer tissue is significantly higher than without lymph node metastasis group and high expression of VEGF-C may be important factors lead to the early tongue cancer lymph node metastasis;4, P16 methylation and expression of VEGF-C in tongue cancer lymphatic metastasis has obvious relevance, P16 methylation can be used for early detection and diagnosis of tongue cancer susceptible important judgment of VEGF-C invasive and can be used as a clinical judgment tongue cancer metastasis potential and prognosis of important reference indicator;... |
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