趋化因子受体CXCR3及其相关MircoRNAs在涎腺肿瘤组织中的表达研究

发布时间：2018-05-06 11:57

本文选题：涎腺肿瘤 + 趋化因子受体CXCR3　；参考：《昆明医科大学》2016年硕士论文

【摘要】：[目的]探讨趋化因子受体CXCR3及其相关MircoRNAs在涎腺肿瘤组织中的表达,分析趋化因子受体CXCR3与相关MircoRNAs、微血管密度(MVD)的相关性以及其在涎腺肿瘤发生发展中的作用。[方法](1)选取术后口腔颌面部涎腺肿瘤组织石蜡包埋标本47例,其中涎腺腺样囊性癌28例,腮腺多形性腺瘤19例,并以19例腮腺多形性腺瘤配对的肿瘤组织旁的正常腮腺组织为对照。(2)对纳入研究的颌面部涎腺肿瘤组织常规行病理切片HE染色,以进行病例确诊和组织分型。(3) EnVision免疫组织化学法检测趋化因子受体CXCR3的表达。光镜下阅片,以胞质或胞浆内出现棕黄色颗粒为阳性着色,每张切片随机观察5个高倍视野,半定量积分法判断结果。具体标准为：①阳性细胞数：≤5%为0分,6%～25%为1分,26%～50%为2分,51%～75%为3分,75%为4分；②阳性强度(以多数细胞判定)：淡黄色为1分,黄色为2分,棕黄色为3分。以①、②两者的乘积判断阴阳性等级：0分为阴性(-),1～4分为弱阳性(+),5～8分为阳性(++),9～12分为强阳性(+++)。两人双盲法观察切片,若两人结果相差3分则重新判断。(4)微血管密度的测量采用CD34免疫组织化学染色,在光镜下以细胞胞膜、胞浆呈棕黄色或黄色颗粒着色判读为CD34阳性染色,每张切片随机观察3个高倍视野,应用Image-pro Plus 6.0专业图像分析软件对其进行半定量分析。(5)应用Targetscan软件预测与趋化因子受体CXCR3相关的MircoRNAs,选取其中与CXCR3相关度较高的miR-486-3p、 miR-149-3p及miR-122-3p进行表达分析。对石蜡包埋的各例组织进行连续切片,抽提其RNA并质检后,采用Realtime PCR方法进行扩增检测miR-486-3p、 miR-149-3p及miR-122-3p在组织中的表达量,分析它们与趋化因子受体CXCR3表达的相关性。(7)数据统计学分析采用SPSS20.0统计软件进行。[结果](1)腮腺组织存在CXCR3表达。正常腮腺组CXCR3表达结果为阴性(-)占52.63%,弱阳性(+)占31.58%,阳性(++)占10.53%,强阳性(+++)占5.26%。多形性腺瘤组CXCR3表达结果为阴性(-)占5.26%,弱阳性(+)占5.26%,阳性(++)占52.63%,强阳性(+++)占36.85%。腺样囊性癌组CXCR3表达结果为阴性(-)占7.14%,弱阳性(+)占17.86%,阳性(++)占17.86%,强阳性(+++)占57.14%。正常腮腺组、多形性腺瘤组及腺样囊性癌组中CXCR3表达强度差异有统计学意义(P0.001)；进一步行Z检验,除多形性腺瘤组和腺样囊性癌组无统计学差异,其余各组间均有统计学差异(P0.05)。(2)正常腮腺组微血管密度为(0.28+0.07),多形性腺瘤组和涎腺腺样囊性癌组微血管密度分别为(0.42±0.12)和(0.49±0.09)。经统计学分析,三组组织微血管密度差异均有统计学差异(P0.001),进一步行LSD-t检验,三组组间差异均有统计学意义(P0.05)。经Spearman相关性分析发现组织中微血管密度(MVD)与趋化因子受体CXCR3的表达存在正相关关系(r=0.419, P0.001)。(3)RealtimePCR检测发现,miR-486-3p在正常腮腺组中阳性率为68.4%,在多形性腺瘤组中为31.6%,在腺样囊性癌组中为32.1%,三组间比较表达差异有统计学意义(P=0.025),进一步行Z检验,除多形性腺瘤组和腺样囊性癌组无统计学差异,其余各组间均有统计学差异(P0.05)。miR-149-3p在正常腮腺组中阳性率为47.4%,在多形性腺瘤组中为26.3%,在腺样囊性癌组中为39.3%,三组间比较表达差异无统计学意义(P=0.400)。miR-122-3p在正常腮腺组中阳性率为11%,在多形性腺瘤组中为11%,在腺样囊性癌组中10.7%,三组间比较表达差异无统计学意义(P=1.00)。经Spearman相关性分析发现组织中miR-486-3p与趋化因子受体CXCR3的表达存在负相关关系(r=-0.302,P=-0.013)。[结论]趋化因子受体CXCR3在涎腺肿瘤组织中的表达明显高于正常组织,而miR-486-3p的表达水平明显低于正常组织,CXCR3的表达与miR-486-3p的表达呈负相关关系而与微血管密度(MVD)呈正相关关系,提示miR-486-3p的下调可能提高CXCR3表达从而诱导涎腺肿瘤血管生成,促进肿瘤发生发展。
[Abstract]:[Objective] to investigate the expression of chemokine receptor CXCR3 and its associated MircoRNAs in salivary gland tumor tissue, and to analyze the correlation between chemokine receptor CXCR3 and related MircoRNAs, microvascular density (MVD) and its role in the occurrence and development of salivary gland tumors. [method] (1) to select paraffin embedded specimens of oral and maxillofacial salivary gland tumor after operation (47) For example, 28 cases of salivary adenoid cystic carcinoma, 19 cases of parotid pleomorphic adenoma, and normal parotid gland adjacent to the tumor tissues of 19 parotid pleomorphic adenomas were compared. (2) routine pathological section of HE staining of the salivary gland tumor tissue of the study was performed to make the diagnosis and tissue typing. (3) EnVision immunohistochemical method. The expression of chemokine receptor CXCR3 was detected. Under the light microscope, the brown yellow granules were stained in cytoplasm or cytoplasm. Each slice was randomly observed with 5 high times of visual field and semi quantitative integral method to determine the results. The specific criteria were: (1) the number of positive cells was 0, 6% to 25% was 1, 26% to 50% was 2, 51% ~ 75% was 3, 75% was 4. (2) positive intensity (determined by most cells): light yellow was 1, yellow was 2, and Brown was 3. To judge Yin and Yang by the product of the two, 0 is negative (-), 1~4 is weak positive (+), 5~8 is positive (+ +), 9~12 is strong positive (+ + +). Two people double blind method to observe slice, if two people difference 3 is rejudged 3. (4) the microvascular density was measured by CD34 immunohistochemical staining. The cell membrane was used under the light microscope, the cytoplasm was stained with brown yellow or yellow granules, and the CD34 positive staining was read. Each slice was randomly observed 3 high times of visual field, and the Image-pro Plus 6 professional image analysis soft parts were used to semi quantitative analysis. (5) the application of Targetscan software The MircoRNAs related to chemokine receptor CXCR3 was predicted, and miR-486-3p, miR-149-3p and miR-122-3p, which had higher correlation with CXCR3, were selected for the expression analysis. The samples of paraffin embedded tissues were sectioned continuously, and the RNA was extracted and the Realtime PCR method was used to detect miR-486-3p, miR-149-3p and miR-122-3p. Expression in the tissue and the correlation between them and the expression of chemokine receptor CXCR3. (7) statistical analysis of data was carried out by SPSS20.0 statistical software. [results] (1) there was CXCR3 expression in parotid gland. The expression of CXCR3 in parotid gland was negative (-) 52.63%, weak positive (+) accounted for 31.58%, positive (+ +) accounted for 10.53%, and strong positive (+ + +) accounted for more than 5.26%. The expression of CXCR3 in the formatting adenoma group was negative (-) 5.26%, weak positive (+) accounted for 5.26%, positive (+ +) accounted for 52.63%, strong positive (+ + +) in the 36.85%. adenoid cystic carcinoma group was negative (-) 7.14%, weak positive (+) accounted for 17.86%, positive (+ +) accounted for 17.86%, strong positive (+ + +) accounted for the normal parotid gland group of 57.14%., pleomorphic adenoma group and adenoid cystic carcinoma group C The difference of XCR3 expression intensity was statistically significant (P0.001); there was no statistical difference between the pleomorphic adenoma group and the adenoid cystic carcinoma group (P0.05). (2) the microvascular density in the normal parotid group was (0.28+0.07), and the microvascular density in the pleomorphic adenoma group and the salivary adenoid cystic carcinoma group was (0.42). The differences of microvascular density in the three groups were statistically different (P0.001). The difference between the three groups was statistically significant (P0.001). The difference between the three groups was statistically significant (P0.05). The correlation between the microvascular density (MVD) and the expression of chemokine receptor CXCR3 was found to be positively correlated with the Spearman correlation analysis (r=0.4). 19, P0.001) (3) RealtimePCR detection found that the positive rate of miR-486-3p in the normal parotid group was 68.4%, 31.6% in the pleomorphic adenoma group, 32.1% in the adenoid cystic carcinoma group, and the difference between the three groups was statistically significant (P=0.025), and the Z test was further performed with no statistical difference between the pleomorphic adenoma group and the adenoid cystic carcinoma group. The positive rate of.MiR-149-3p in the normal parotid group was 47.4%, 26.3% in the pleomorphic adenoma group and 39.3% in the adenoid cystic carcinoma group. There was no statistical difference between the three groups (P=0.400), the Zhongyang sex rate was 11% in the normal parotid group and 11% in the pleomorphic adenoma group, and in the adenoid cystic carcinoma group. There was no significant difference in the expression of the 10.7% and three groups in the cancer group (P=1.00). The expression of miR-486-3p and chemokine receptor CXCR3 in the tissue was negatively correlated (r=-0.302, P=-0.013). [Conclusion] the expression of chemokine receptor CXCR3 in salivary gland tumor tissues was significantly higher than that of normal tissue, while miR-486-3p was significantly higher than that of normal tissue. The expression level of the CXCR3 was significantly lower than that of the normal tissue. The expression of miR-486-3p was negatively correlated with the microvascular density (MVD), suggesting that the downregulation of miR-486-3p might increase the expression of CXCR3 and induce the angiogenesis of salivary tumor and promote the development of the tumor.

【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R739.8

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