本文選題：涎腺腫瘤 + 趨化因子受體CXCR3��； 參考：《昆明醫(yī)科大學(xué)》2016年碩士論文

【摘要】：[目的]探討趨化因子受體CXCR3及其相關(guān)MircoRNAs在涎腺腫瘤組織中的表達(dá),分析趨化因子受體CXCR3與相關(guān)MircoRNAs、微血管密度(MVD)的相關(guān)性以及其在涎腺腫瘤發(fā)生發(fā)展中的作用。[方法](1)選取術(shù)后口腔頜面部涎腺腫瘤組織石蠟包埋標(biāo)本47例,其中涎腺腺樣囊性癌28例,腮腺多形性腺瘤19例,并以19例腮腺多形性腺瘤配對(duì)的腫瘤組織旁的正常腮腺組織為對(duì)照。(2)對(duì)納入研究的頜面部涎腺腫瘤組織常規(guī)行病理切片HE染色,以進(jìn)行病例確診和組織分型。(3) EnVision免疫組織化學(xué)法檢測(cè)趨化因子受體CXCR3的表達(dá)。光鏡下閱片,以胞質(zhì)或胞漿內(nèi)出現(xiàn)棕黃色顆粒為陽(yáng)性著色,每張切片隨機(jī)觀察5個(gè)高倍視野,半定量積分法判斷結(jié)果。具體標(biāo)準(zhǔn)為：①陽(yáng)性細(xì)胞數(shù)：≤5%為0分,6%～25%為1分,26%～50%為2分,51%～75%為3分,75%為4分；②陽(yáng)性強(qiáng)度(以多數(shù)細(xì)胞判定)：淡黃色為1分,黃色為2分,棕黃色為3分。以①、②兩者的乘積判斷陰陽(yáng)性等級(jí)：0分為陰性(-),1～4分為弱陽(yáng)性(+),5～8分為陽(yáng)性(++),9～12分為強(qiáng)陽(yáng)性(+++)。兩人雙盲法觀察切片,若兩人結(jié)果相差3分則重新判斷。(4)微血管密度的測(cè)量采用CD34免疫組織化學(xué)染色,在光鏡下以細(xì)胞胞膜、胞漿呈棕黃色或黃色顆粒著色判讀為CD34陽(yáng)性染色,每張切片隨機(jī)觀察3個(gè)高倍視野,應(yīng)用Image-pro Plus 6.0專業(yè)圖像分析軟件對(duì)其進(jìn)行半定量分析。(5)應(yīng)用Targetscan軟件預(yù)測(cè)與趨化因子受體CXCR3相關(guān)的MircoRNAs,選取其中與CXCR3相關(guān)度較高的miR-486-3p、 miR-149-3p及miR-122-3p進(jìn)行表達(dá)分析。對(duì)石蠟包埋的各例組織進(jìn)行連續(xù)切片,抽提其RNA并質(zhì)檢后,采用Realtime PCR方法進(jìn)行擴(kuò)增檢測(cè)miR-486-3p、 miR-149-3p及miR-122-3p在組織中的表達(dá)量,分析它們與趨化因子受體CXCR3表達(dá)的相關(guān)性。(7)數(shù)據(jù)統(tǒng)計(jì)學(xué)分析采用SPSS20.0統(tǒng)計(jì)軟件進(jìn)行。[結(jié)果](1)腮腺組織存在CXCR3表達(dá)。正常腮腺組CXCR3表達(dá)結(jié)果為陰性(-)占52.63%,弱陽(yáng)性(+)占31.58%,陽(yáng)性(++)占10.53%,強(qiáng)陽(yáng)性(+++)占5.26%。多形性腺瘤組CXCR3表達(dá)結(jié)果為陰性(-)占5.26%,弱陽(yáng)性(+)占5.26%,陽(yáng)性(++)占52.63%,強(qiáng)陽(yáng)性(+++)占36.85%。腺樣囊性癌組CXCR3表達(dá)結(jié)果為陰性(-)占7.14%,弱陽(yáng)性(+)占17.86%,陽(yáng)性(++)占17.86%,強(qiáng)陽(yáng)性(+++)占57.14%。正常腮腺組、多形性腺瘤組及腺樣囊性癌組中CXCR3表達(dá)強(qiáng)度差異有統(tǒng)計(jì)學(xué)意義(P0.001)；進(jìn)一步行Z檢驗(yàn),除多形性腺瘤組和腺樣囊性癌組無(wú)統(tǒng)計(jì)學(xué)差異,其余各組間均有統(tǒng)計(jì)學(xué)差異(P0.05)。(2)正常腮腺組微血管密度為(0.28+0.07),多形性腺瘤組和涎腺腺樣囊性癌組微血管密度分別為(0.42±0.12)和(0.49±0.09)。經(jīng)統(tǒng)計(jì)學(xué)分析,三組組織微血管密度差異均有統(tǒng)計(jì)學(xué)差異(P0.001),進(jìn)一步行LSD-t檢驗(yàn),三組組間差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。經(jīng)Spearman相關(guān)性分析發(fā)現(xiàn)組織中微血管密度(MVD)與趨化因子受體CXCR3的表達(dá)存在正相關(guān)關(guān)系(r=0.419, P0.001)。(3)RealtimePCR檢測(cè)發(fā)現(xiàn),miR-486-3p在正常腮腺組中陽(yáng)性率為68.4%,在多形性腺瘤組中為31.6%,在腺樣囊性癌組中為32.1%,三組間比較表達(dá)差異有統(tǒng)計(jì)學(xué)意義(P=0.025),進(jìn)一步行Z檢驗(yàn),除多形性腺瘤組和腺樣囊性癌組無(wú)統(tǒng)計(jì)學(xué)差異,其余各組間均有統(tǒng)計(jì)學(xué)差異(P0.05)。miR-149-3p在正常腮腺組中陽(yáng)性率為47.4%,在多形性腺瘤組中為26.3%,在腺樣囊性癌組中為39.3%,三組間比較表達(dá)差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.400)。miR-122-3p在正常腮腺組中陽(yáng)性率為11%,在多形性腺瘤組中為11%,在腺樣囊性癌組中10.7%,三組間比較表達(dá)差異無(wú)統(tǒng)計(jì)學(xué)意義(P=1.00)。經(jīng)Spearman相關(guān)性分析發(fā)現(xiàn)組織中miR-486-3p與趨化因子受體CXCR3的表達(dá)存在負(fù)相關(guān)關(guān)系(r=-0.302,P=-0.013)。[結(jié)論]趨化因子受體CXCR3在涎腺腫瘤組織中的表達(dá)明顯高于正常組織,而miR-486-3p的表達(dá)水平明顯低于正常組織,CXCR3的表達(dá)與miR-486-3p的表達(dá)呈負(fù)相關(guān)關(guān)系而與微血管密度(MVD)呈正相關(guān)關(guān)系,提示miR-486-3p的下調(diào)可能提高CXCR3表達(dá)從而誘導(dǎo)涎腺腫瘤血管生成,促進(jìn)腫瘤發(fā)生發(fā)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.

【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R739.8

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