The Expression And Significance Of EphA2, HoxA1 And C-myc In Laryngeal Carcinoma

Objective: The genesis and development of The carcinoma of larynx concern changes of regulatory genes and signal transduction. Eph receptors and their ligands Ephrins can regulate cell growth and development. As a regulatory gene family, Hox gene not only regulate generation and different- iation of the embryo, but also regulate generation and differentiation of the cells in adult. The anomal-expression of the Hox gene induces anomal-appearance of an individual, Furthermore, it can induce malignant transformation in cells. The c-myc oncogene is one of neucleoprotein gene, It plays an important role in tumorigenesis, malignant transformation of the cells, differentiation and Apoptosis of the cells, and so on. The expression of EphA2 can be regulated by many genes such as ras, p53, HoxA1, HoxB1, Estrogen receptor and c-myc. In our study, our aim is to investigate the correlation among the expression of EphA2, Hoxa1 and c-myc and the development of the cancer of larynx. We try to explore genesis mechanism of laryngeal carcinoma at the level of molecular biology.Methods: 1. 40 laryngeal carcinoma fresh samples were analyzed and meanwhile 40 para-carcinoma tissues (to cutting margin>0.5cm) and 15 normal laryngeal mucosa samples were also studied as controls. All subjects are taken from the patients who accepted operation treatment in the Fourth Hospital of HeBei Medical University from 2007.1 to 2008.12. All tumor tissues are confirmed to be squamous-cell carcinoma; the para- carcinoma tissues and normal mucosa are confirmed to be inflammatory or normal mucosa pathologically. 2.We measured the expression of EphA2, Hoxa1 and c-myc protein with the Flow Cytometere (Epics-XLⅡ). Fluorescence Index was defined as the quantitative expression index of EphA2, Hoxa1 and c-myc protein. The FI value more than 1.0 should be considered as positive expression, otherwise to be negative expression. 3. We measured the expression of EphA2, Hoxa1 and c-myc protein by immunohistochemical staining,. Positive stain of EphA2 protein was located in cytoplasm and membrane, Positive stain of Hoxa1 protein was located in cytoplasm. The scoring scheme was based on both percentage of positive cell and staining intensity."0"score was considered as normal expression. more than 0 score was considered as abnormal expression. Positive stain of c-myc was located in cytoplasm and nuclei. more than 1 score was considered as abnormal expressions. 4. Statistic analysis was performed using the SPSS 11.5 for windows software, P≤0.05 indicating significant difference, P≤0.01 indicating very significant difference .Results: FCM:1 The quantitative (FI=1.4877) and qualitative (77.5%) expression of EphA2 protein in laryngeal carcinoma tissues is obviously higher than those in para-carcinoma(1.2178, 55%)and in normal laryngeal mucosa(0.9999, 40%)tissues respectively, (P<0.05) (χ2=4.528, P=0.033); (P<0.05) (χ2=5.369, P=0.021); There is no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues. In laryngeal carcinoma, the quantitative and qualitative expression of EphA2 protein with positive metastasis(1.8264, 100%) is remarkably higher than those with negative metastasis(1.3592,67.9%) respectively, (P=0.002) (Fisher's exact test, P=0.037); in clinical stageⅢ,Ⅳ(1.6757, 91.7%) higher than in clinical stageⅠ,Ⅱ(1.2579, 56.3%) (P=0.002) (Fisher's exact test, P=0.018); in pathological gradeⅢthe quantitative expression(1.8110) higher than in pathological gradeⅠ(1.3871), (P < 0.05), but there is no significant difference between pathological gradeⅠandⅡ(1.7675), neither pathologicalⅡandⅢ. There is no significant difference among the qualitative expression in pathological gradeⅠ(72.4%),Ⅱ(85.7%) andⅢ(100%). The expression of EphA2 protein isn't significantly related to patients'clinical classification, tumor size, smoking history, patients'age .2 The quantitative (FI=1.1810) and qualitative (90%) expression of Hoxa1 protein in laryngeal carcinoma tissues is obviously higher than those in para-carcinoma (0.9743,27.5%) and in normal laryngeal mucosa tissues (0.9999, 26.7%) respectively, (P<0.05) (χ2=32.2373,P<0.005); (P<0.05) (χ2=21.9841,P<0.005); There is no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues. There is no significant difference between the quantitative expression of Hoxa1 protein with positive metastasis(1.1823) and those with negative metastasis(1.1225), the qualitative expression (91.7%, 89.3%) has no difference; The quantitative expression has no significant difference among clinical stageⅢ,Ⅳ(1.1941) and clinical stageⅠ,Ⅱ(1.1291), but the qualitative expression of clinical stageⅢ,Ⅳis obviously higher than those in clinical stageⅠ,Ⅱ(P=0.02);there is no significant difference between pathological gradeⅠandⅡandⅢ. The expression of Hoxa1 protein isn't significantly related to patients'clinical classification, tumor size, smoking history, patients'age .3 The quantitative (FI=1.1741) and qualitative (72.5%) expression of c-myc protein in laryngeal carcinoma tissues is obviously higher than those in para-carcinoma (0.9895,45%) and in normal laryngeal mucosa tissues(1.0000, 40%) respectively, (P < 0.05) (χ2 =6.240 , P=0.012); (P < 0.05) (χ2=4.979,P=0.026); There is no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues. the quantitative expression of c-myc protein with positive metastasis(1.1125) is lower than those with negative metastasis(1.2136), (t=2.681,P=0.011), There has significant difference between the qualitative expression(85.7%, 41.7%) (Fisher's exact test, P=0.008); There is no significant difference between clinical stageⅢ,Ⅳ(1.1713 , 75%) and clinical stageⅠ,Ⅱ(1.1663, 68.8%); there is no significant difference among pathological gradeⅠ(1.1676,75.9%)andⅡ(1.1914,71.4%)andⅢ(1.2051,50%). The expression of c-myc protein isn't significantly related to patients'clinical classification, tumor size, smoking history, patients'age .4 There is multiple linear regression relationship in the expression of EphA2, Hoxa1 and c-myc. In laryngeal carcinoma, and it is a positive correlation. The multiple linear regression equation is Y=0.983+0.739X1-0.323X2(P=0.000)。Immunohistochemistry1. The percentage of abnormal expression of EphA2 in laryngeal carcinoma(80%) is significantly higher than that(40%) in laryngeal mucous(P=0.008). so is between clinical stageⅢ,Ⅳand clinical stageⅠ,Ⅱ(P=0.004). But there is no relationship between EphA2 and other clinical factors such as size of tumor, metastasis.2 The percentage of abnormal expression of HoxA1 in laryngeal carcinoma(82.5%) is significantly higher than that(33.3%) in laryngeal mucous(P=0.001). so is between clinical stageⅢ,Ⅳ(95.8%) and clinical stageⅠ,Ⅱ(62.5%) (P=0.011). But there is no relationship between HoxA1 and other clinical factors such as size of tumor, metastasis . 3 The percentage of abnormal expression of c-myc in laryngeal carcinoma(77.5%) is significantly higher than that(20%) in laryngeal mucous(P=0.000). so is between metastasis and non-metastasis(P=0.012). But there is no relationship between c-myc and other clinical factors such as size of tumor, clinical stage.4 Spearman rank correlation showes the abnormal expression of EphA2 is related to that of HoxA1 (rs=0.420, P=0.007), so is between EphA2 and c-myc (rs=-0.395 , P=0.012).Conclusions:1 EphA2, Hoxa1 and C-myc protein express in all laryngeal carcinoma, para-carcinoma mucosa and normal mucosa tissues.2 The quantitative and qualitative expression of EphA2, Hoxa1 and C-myc protein in laryngeal carcinoma tissues is obviously higher than those in para-carcinoma and in normal laryngeal mucosa tissues respectively. There is no significant difference between the expression of para-carcinoma and normal laryngeal mucosa tissues.3 The expression of EphA2 isn't related to patients'clinical classification, tumor size, smoking history, patients'age, but related to lymph nodes metastasis, clinical stage and pathological grade. The expression of Hoxa1 isn't related to patients'clinical classification, tumor size, smoking history, patients'age, lymph nodes metastasis and pathological grade. but related to clinical stage. The expression of C-myc isn't related to patients'clinical classification, tumor size, smoking history, patients'age, clinical stage and pathological grade, but related to lymph nodes metastasis.4 There is multiple linear regression relationship in the expression of EphA2, Hoxa1 and C-myc In laryngeal carcinoma, it is a positive correlation. The expression of Hoxa1 increases along with that of EphA2. The expression of C-myc decreases along with accrescence that of EphA2.5 The expression of EphA2, Hoxa1 and C-myc may contribute to the carcinogenesis and development of laryngeal carcinoma.