The Expression Research Of Chemokine Receptor CXCR3 And Its Related MircoRNAs In Salivary Gland Tumor Tissues

[Objective]To investigate the expression of chemokine receptor CXCR3 and MircoRNAs in salivary gland tumor tissues. Analysis the relationship between chemokine receptor CXCR3 with MircoRNAs, micro vessel density (MVD) and the effects in the tumor of maxillofacial salivary gland.[Methods] (1) Select 47 cases specimens of paraffin-embedded tumor tissue of salivary gland in oral and maxillofacial, including 28 cases of adenoid cystic carcinoma,19 cases pleomorphic adenoma, and 19 cases of parotid pleomorphic adenoma pairing with normal parotid gland to control. (2) The salivary gland tumor tissues were examined with haematoxylin eosin staining. (3) The expressions of chemokine receptor CXCR3 were detected by EnVision immunohistochemical method. The brown granules in cell membrane or cytoplasm were considered positive stained cell; Fiver fields in each sample were observed randomly with microscope to determine expression level of CXCR3. The criteria was:① The number of positive cells≤5% scored 0 points,6%～25% scored 1 points,26%～50% scored 2 points, 51%～75% scored 3 points,>75% scored 4 points; ② Positive intensity:Yellowish scored 1 point, yellow scored 2 points, brown scored 3 points. To product ① with ② to judge the level of positive or negative:0 point scored negative (-),1-4 points scored weakly positive (+),5-8 points scored positive (++),9-12 points scored strong positive (+++). (4) The micro vascular density was measured by immunohistochemical staining of CD34. Under microscope, the cell was considered positive staining of CD34 when there were yellow or brown yellow particles in its membrane or cytoplasm. Semi quantity of microvascular density was analyzed with Image analysis software (Image-pro Plus 6.0). (5) The MircoRNAs related to chemokine receptor CXCR3 were predicted by the target-scan software,there was closed correlation between CXCR3 and miR-486-3p, miR-149-3p and miR-122-3p. (6) The paraffin embedded tissues were serially sectioned and extracted RNA. The expression levels of miR-486-3p, miR-149-3p and miR-122-3p in the tissues were detected by Real-time PCR. (7) The relationships between CXCR3 and miR-486-3p, miR-149-3p and miR-122-3p were analyzed with SPSS 20.0 statistical software.[Results](1) There was CXCR3 expression in parotid gland. In normal parotid group, the negative (-) expression of CXCR3 was 52.63%, weak positive (+) was 31.58%, positive (++) was 10.53%, strong positive (+++) was 5.26%. In pleomorphic adenoma, the negative (-) expression of CXCR3 was 5.26%, weak positive (+) was 5.26%, positive (++) was 52.36%, strong positive (+++) was 36.85%. In adenoid cystic carcinoma group, the negative (-) expression of CXCR3 was 7.14%, weak positive (+) was 17.86%, positive (++) was 17.86%, strong positive (++ +) was 57.14%. There were significant differences in the expression intensity of CXCR3 in normal parotid gland, polymorphic adenoma group and adenoid cystic carcinoma group (P<0.001); Through Z test, in addition to polymorphic adenoma and adenoid cystic carcinoma group, there were significant differences between the other groups (P<0.05). (2) The microvessel density of normal parotid gland was (0.28 ± 0.07), the group of polymorphic adenoma was (0.42±0.12) and adenoid cystic carcinoma was (0.49±0.09). The microvessel density between these three groups,there were significant differences (P<0.001), the difference was significant between three groups by Z test (P<0.05). Through Spearman correlation analysis revealed that there was a positive correlation between microvessel density (MVD) with the expression of chemokine receptor CXCR3 (r=0.419, P<0.001). (3) Realtime PCR detection revealed that the positive rate of miR-486-3p in normal parotid group was 68.4%(13/19). In pleomorphic adenoma group, the positive rate was 31.6% (6/19). In adenoid cystic carcinoma group,the positive rate was 32.1% (9/28). There were significant differences between two groups (P=0.025). in addition to the polymorphic adenoma and adenoid cystic carcinoma group, there were significant differences in another groups (P<0.05); The positive rate of MiR-149-3p in normal parotid group was 47.4% (9/19). In pleomorphic adenoma was 26.3% (5/19). In adenoid cystic carcinoma group the positive rate was 39.3%(11/28). There were no difference between each group (P=0.400). The positive rate of MiR-122-3pwas 11% (2/19), in normal parotid group. In pleomorphic adenoma group was The positive rate of MiR-122-3p11%(2/19). In adenoid cystic carcinoma group, the positive rate was 10.7%(3/28). There were no differences between each group(P=1.00). Spearman correlation analysis revealed that there was a negative correlation between the expression of miR-486-3p and chemokine receptor CXCR3(r=-0.302, P=0.013).[Conclusions] The expression of chemokine receptor CXCR3 in salivary gland was higher than in normal tissues significantly, However, the expression of miR-486-3p was lower than the normal tissue significantly; The expression of CXCR3 showed a negative correlation with miR-486-3p and show positive correlation with Microvessel density (MVD), Above all revealed that down-regulation of miR-486-3p may enhanced the expression of CXCR3 and to induce angiogenesis in salivary gland and promote the tumor occurrence and development.