SLIT2、SOX1及JAM3基因甲基化水平在宫颈癌发生中的临床意义

发布时间：2018-04-27 11:55

本文选题：宫颈癌 + 宫颈上皮内病变　；参考：《天津医科大学》2017年硕士论文

【摘要】：目的:应用焦磷酸测序技术定量检测高危型人乳头瘤病毒感染的不同宫颈病变程度的脱落细胞中SLIT2、SOX1和JAM3基因的甲基化水平,比较各基因在各组间甲基化水平的差异,分析其在宫颈癌发生、发展过程中的作用,并探究它们在宫颈癌早期筛查中的临床价值。方法:收取由HCII证实的Hr-HPV感染并经组织病理学诊断的不同宫颈病变程度的脱落细胞,包括正常组45份,宫颈上皮内病变I级(CINI)47份,宫颈上皮内病变II/III级(CINII/III)70份,宫颈癌组85份。采用亚硫酸氢盐修饰结合焦磷酸测序技术检测宫颈脱落细胞中SLIT2、SOX1和JAM3基因的甲基化水平,评价它们对于高级别宫颈癌前病变及宫颈癌的诊断价值。使用SPSS20.0软件对资料进行处理和分析,甲基化的平均水平用平均数±标准差(`X±S)表示,应用单因素方差分析处理数据,组间多重比较采用LSD方法,制作受试者的工作特征曲线(receiver operating characteristic curve,ROC),并计算曲线下面积(AUC),取得最佳诊断阈值。结果:1.正常宫颈组、CINI组、CINII/III组以及宫颈癌组的脱落细胞标本中SLIT2甲基化水平分别为(5.29±1.29)%、(5.75±2.57)%、(11.65±6.74)%、(25.22±14.63)%,组间差异有统计学意义(F=66.72,P0.001)。SOX1甲基化水平分别为(5.78±1.17)%、(5.35±1.36)%、(13.17±8.42)%、(32.30±16.38)%,组间差异有统计学意义(F=96.81,P0.001)。JAM3甲基化水平分别为(3.84±1.84)%、(3.96±1.79)%、(13.78±7.06)%、(25.58±13.09)%,组间差异具有统计学意义(F=92.77,P0.001)。结果显示各基因的宫颈癌组甲基化水平明显高于其他各组(P0.001),并且甲基化水平随宫颈病变程度的加重而升高。2.定量检测SLIT2、SOX1和JAM3基因甲基化水平诊断宫颈癌的ROC曲线下面积分别为0.914、0.953和0.925。经ROC曲线下面积计算获得最佳SLIT2基因甲基化水平的界定值为14.08%,敏感性为74.0%,特异性为90.0%。当SOX1界定值为17.25%,敏感性为85.0%,特异性为90.0%。当JAM3基因界定值为16.13%,敏感性为81.0%,特异性为90.0%。同样,诊断CINII/III+的ROC曲线下面积分别为0.940、0.933和0.982。计算获得最佳SLIT2基因甲基化水平的界定值为7.08%,敏感性为86.5%,特异性为88.0%。当SOX1界定值为9.58%,敏感性为83.2%,特异性为100.0%。当JAM3界定值为10.37%,敏感性为85.8%,特异性为100.0%。3.将SLIT2、SOX1及JAM3三基因进行两两联合,发现SLIT2/SOX1诊断宫颈癌的敏感性和特异性分别为82.9%和88.0%,CINII/III+的敏感性和特异性为85.8%和90.8%;SLIT2/JAM3诊断宫颈癌的敏感性和特异性分别为87.1%和86.7%,CINII/III+的敏感性和特异性为90.0%和89.7%;SOX1/JAM3诊断宫颈癌的敏感性和特异性分别为82.4%和91.0%,CINII/III+的敏感性和特异性为90.0%和92.9%。结论:1.在宫颈癌中存在抑癌基因SLIT2、SOX1及JAM3的高甲基化现象,并且甲基化水平随着宫颈病变程度的加重而升高;2.定量检测抑癌基因SLIT2、SOX1及JAM3的甲基化水平对于诊断宫颈癌及高级别癌前病变具有较高的敏感性和特异性;3.联合基因检测对于宫颈癌及高级别癌前病变具有较高的诊断效能;4.焦磷酸测序技术具有操作简单、准确性高、重现性好、高通量且低成本的优点,适用于宫颈癌的早期筛查。
[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.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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