本文選題：宮頸癌 + 宮頸上皮內(nèi)病變　； 參考：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:應(yīng)用焦磷酸測序技術(shù)定量檢測高危型人乳頭瘤病毒感染的不同宮頸病變程度的脫落細(xì)胞中SLIT2、SOX1和JAM3基因的甲基化水平,比較各基因在各組間甲基化水平的差異,分析其在宮頸癌發(fā)生、發(fā)展過程中的作用,并探究它們在宮頸癌早期篩查中的臨床價(jià)值。方法:收取由HCII證實(shí)的Hr-HPV感染并經(jīng)組織病理學(xué)診斷的不同宮頸病變程度的脫落細(xì)胞,包括正常組45份,宮頸上皮內(nèi)病變I級(CINI)47份,宮頸上皮內(nèi)病變II/III級(CINII/III)70份,宮頸癌組85份。采用亞硫酸氫鹽修飾結(jié)合焦磷酸測序技術(shù)檢測宮頸脫落細(xì)胞中SLIT2、SOX1和JAM3基因的甲基化水平,評價(jià)它們對于高級別宮頸癌前病變及宮頸癌的診斷價(jià)值。使用SPSS20.0軟件對資料進(jìn)行處理和分析,甲基化的平均水平用平均數(shù)±標(biāo)準(zhǔn)差(`X±S)表示,應(yīng)用單因素方差分析處理數(shù)據(jù),組間多重比較采用LSD方法,制作受試者的工作特征曲線(receiver operating characteristic curve,ROC),并計(jì)算曲線下面積(AUC),取得最佳診斷閾值。結(jié)果:1.正常宮頸組、CINI組、CINII/III組以及宮頸癌組的脫落細(xì)胞標(biāo)本中SLIT2甲基化水平分別為(5.29±1.29)%、(5.75±2.57)%、(11.65±6.74)%、(25.22±14.63)%,組間差異有統(tǒng)計(jì)學(xué)意義(F=66.72,P0.001)。SOX1甲基化水平分別為(5.78±1.17)%、(5.35±1.36)%、(13.17±8.42)%、(32.30±16.38)%,組間差異有統(tǒng)計(jì)學(xué)意義(F=96.81,P0.001)。JAM3甲基化水平分別為(3.84±1.84)%、(3.96±1.79)%、(13.78±7.06)%、(25.58±13.09)%,組間差異具有統(tǒng)計(jì)學(xué)意義(F=92.77,P0.001)。結(jié)果顯示各基因的宮頸癌組甲基化水平明顯高于其他各組(P0.001),并且甲基化水平隨宮頸病變程度的加重而升高。2.定量檢測SLIT2、SOX1和JAM3基因甲基化水平診斷宮頸癌的ROC曲線下面積分別為0.914、0.953和0.925。經(jīng)ROC曲線下面積計(jì)算獲得最佳SLIT2基因甲基化水平的界定值為14.08%,敏感性為74.0%,特異性為90.0%。當(dāng)SOX1界定值為17.25%,敏感性為85.0%,特異性為90.0%。當(dāng)JAM3基因界定值為16.13%,敏感性為81.0%,特異性為90.0%。同樣,診斷CINII/III+的ROC曲線下面積分別為0.940、0.933和0.982。計(jì)算獲得最佳SLIT2基因甲基化水平的界定值為7.08%,敏感性為86.5%,特異性為88.0%。當(dāng)SOX1界定值為9.58%,敏感性為83.2%,特異性為100.0%。當(dāng)JAM3界定值為10.37%,敏感性為85.8%,特異性為100.0%。3.將SLIT2、SOX1及JAM3三基因進(jìn)行兩兩聯(lián)合,發(fā)現(xiàn)SLIT2/SOX1診斷宮頸癌的敏感性和特異性分別為82.9%和88.0%,CINII/III+的敏感性和特異性為85.8%和90.8%;SLIT2/JAM3診斷宮頸癌的敏感性和特異性分別為87.1%和86.7%,CINII/III+的敏感性和特異性為90.0%和89.7%;SOX1/JAM3診斷宮頸癌的敏感性和特異性分別為82.4%和91.0%,CINII/III+的敏感性和特異性為90.0%和92.9%。結(jié)論:1.在宮頸癌中存在抑癌基因SLIT2、SOX1及JAM3的高甲基化現(xiàn)象,并且甲基化水平隨著宮頸病變程度的加重而升高;2.定量檢測抑癌基因SLIT2、SOX1及JAM3的甲基化水平對于診斷宮頸癌及高級別癌前病變具有較高的敏感性和特異性;3.聯(lián)合基因檢測對于宮頸癌及高級別癌前病變具有較高的診斷效能;4.焦磷酸測序技術(shù)具有操作簡單、準(zhǔn)確性高、重現(xiàn)性好、高通量且低成本的優(yōu)點(diǎn),適用于宮頸癌的早期篩查。
[Abstract]:Objective: to quantify the methylation level of SLIT2, SOX1 and JAM3 genes in exfoliated cells with different cervical lesions of high risk human papillomavirus infection by pyrosequencing, and to compare the difference of methylation levels between each gene in each group, to analyze its role in the development of cervical cancer, and to explore their role in cervical cancer. Clinical value in early screening. Methods: exfoliated cells from HCII confirmed Hr-HPV infection and histopathological diagnosis of different cervical lesions, including 45 normal groups, 47 I (CINI) in cervical intraepithelial lesion, 70 II/III (CINII/III) in cervical intraepithelial lesion and 85 in cervical cancer group. The methylation level of SLIT2, SOX1 and JAM3 genes in cervical exfoliated cells was detected by pyrosequencing, and their diagnostic value for advanced cervical precancerous lesions and cervical cancer was evaluated. SPSS20.0 software was used to process and analyze the data. The average level of methylation was expressed by mean mean standard deviation (`X + S), and the single factor variance score was applied. The LSD method was used to analyze the data. The receiver operating characteristic curve (ROC) was made and the area under the curve (AUC) was calculated. The optimal diagnostic threshold was obtained. Results: 1. the level of SLIT2 methylation in the exfoliated cells of normal cervix, CINI, CINII/III and cervical cancer group. The difference was (5.29 + 1.29)%, (5.75 + 2.57)%, (11.65 + 6.74)% and (25.22 + 14.63)%. The difference between groups was statistically significant (F=66.72, P0.001).SOX1 methylation level was (5.78 + 1.17)%, (5.35 + 1.36)%, (13.17 + 5.75)%, and the difference between groups was statistically significant (F=96.81, P0.001).JAM3 methylation level respectively %, (13.78 + 7.06)%, (25.58 + 13.09)%, the difference between the groups was statistically significant (F=92.77, P0.001). The results showed that the methylation level of the cervical cancer groups of each gene was significantly higher than that of the other groups (P0.001), and the methylation level increased with the degree of cervical lesions and increased by.2. quantitative detection of SLIT2, the methylation level of SOX1 and JAM3 genes in the diagnosis of cervical cancer. The area under the ROC curve is 0.914,0.953 and 0.925. calculated by the area under the ROC curve respectively. The definition of the optimum SLIT2 gene methylation level is 14.08%, the sensitivity is 74%, the specificity is 90.0%. when SOX1 is defined by 17.25%, the sensitivity is 85%, the specificity is 90.0%. when the JAM3 gene is defined 16.13%, the sensitivity is 81%, and the specificity is 90%. Similarly, the area under the ROC curve for the diagnosis of CINII/III+ was 7.08%, the sensitivity was 7.08%, the sensitivity was 86.5%, the specificity was 88.0%. when SOX1 was defined as 9.58%, the sensitivity was 83.2%, the specificity was 100.0%. when JAM3 was 10.37%, the sensitivity was 85.8%, and the specificity was 100%. .3. combined SLIT2, SOX1, and JAM3 three genes to 22. The sensitivity and specificity of SLIT2/SOX1 for diagnosis of cervical cancer were 82.9% and 88% respectively. The sensitivity and specificity of CINII/III+ were 85.8% and 90.8%. The sensitivity and specificity of SLIT2/JAM3 in the diagnosis of cervical cancer were 87.1% and 86.7% respectively. The sensitivity and specificity of CINII/III+ were 90% and 89. .7%; the sensitivity and specificity of SOX1/JAM3 in the diagnosis of cervical cancer were 82.4% and 91% respectively, and the sensitivity and specificity of CINII/III+ were 90% and 92.9%.. 1. in cervical cancer, there was the hypermethylation of tumor suppressor gene SLIT2, SOX1 and JAM3, and the level of methylation increased with the aggravation of cervical disease; 2. quantitative detection of tumor suppressor gene SL The methylation levels of IT2, SOX1 and JAM3 have high sensitivity and specificity for the diagnosis of cervical and advanced precancerous lesions; 3. combined gene detection has a high diagnostic efficiency for cervical and advanced precancerous lesions; 4. pyrosequencing technology has the advantages of simple operation, high accuracy, good reproducibility, high throughput and low cost. It is suitable for early screening of cervical cancer.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.33

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