宫颈癌不同病程组织中HPV16型E2、E6、E7基因的相关研究

发布时间：2018-06-29 09:59

本文选题：HPV16 + E2基因　；参考：《北京协和医学院》2014年硕士论文

【摘要】：本研究选取了HPV16型感染的宫颈癌病例样本(部分含癌旁组织),对E6、E7基因进行测序分析,分别构建进化树,用实时定量PCR的方法检测DNA水平上E2/E6值以判断病毒基因组的存在状态及E2、E6、E7mRNA水平,并分析E6mRNA的剪接情况,研究结果如下： 1.E6和E7的建进化树结果显示只有亚洲变异型和欧洲变异型没有发现非洲1型、非洲2型、亚-美型和北美变异型。检测到T178G-A647G-T846C共突变,其与宫颈癌临床分期没有相关性(x2=0.046,P0.05)。 2.对73例HPV16型病毒感染的样本进行E2和E6基因DNA水平的检测,用比值分析来判定病毒的整合情况。结果显示45例(61.64%)为游离状态,18例(24.66%)为混合状态,10例(13.70%)为整合状态。HPV整合事件与宫颈癌临床分期(x2=0.006,P0.05)和宫颈癌组织学类型(x2=0.623,P0.05)等临床资料之间没有相关性。T178G-A647G-T846C共突变也与病毒的存在状态不相关(x2=3.696,P0.05)。 3.分析E6基因的剪接变化发现,这些样本中E6的剪接异构体主要以E6*I为主(P0.0001),部分样本检测不到E6*I和E6,且E6和E6*I的含量是呈正相关的(r=0.6335,P0.001)。病毒的整合事件(F=0.0667,P0.05)和T178G-A647G-T846C共突变(P0.05)均不影响E6剪接变化。E6剪接变化与宫颈癌的临床分期没有相关性(F=0.6399,P0.05)。 4.E2、E6、E7三个基因的转录本在这些样本中的相对含量不同(F=18.38,P0.001)：E6和E7的转录水平均大于E2的转录水平(P0.001,P0.001),E6和E7的转录水平间无显著性差异(P0.05)。E2基因的转录水平在病毒游离状态高于混合状态(p0.01),游离状态高于整合状态(P0.05)；E6(p0.01)和E7基因(p0.001)的转录水平都是在病毒整合状态高于混合状态(P0.01,P0.001)和游离状态(P0.05,P0.05)。在宫颈癌不同的临床分期中,E2、E6和E7基因的转录水平均无显著性差异(P0.05)。T178G-A647G-T846C共突变不影响E2、E6、E7基因转录本的含量(P0.05)。结论：在本研究涉及的HPV16感染的宫颈癌临床样本范围内,围绕HPV16型的E2、E6和E7三个癌症相关基因所进行的有关检测和分析中,虽然发现这些基因在某些方面存在差异或相关性,但是没有发现与宫颈癌临床分期相关的因素。
[Abstract]:In this study, the samples of cervical cancer infected with HPV16 (including some tissues adjacent to cancer) were selected, and the E6 E7 gene was sequenced, and the evolutionary tree was constructed. The E _ 2 / E _ 6 value at DNA level was detected by real-time quantitative PCR to determine the status of virus genome and the level of E _ 2 / E _ (6) E _ (7) mRNA, and the splicing of E _ (6) mRNA was analyzed. The results are as follows: 1. The results of E6 and E7 show that only Asian and European variants have not found African type 1, African type 2, sub- American variant and North American variant. The comutation of T178G-A647G-T846C was detected, and there was no correlation between T178G-A647G-T846C and the clinical stage of cervical cancer (x20.046). The DNA levels of E2 and E6 genes were detected in 73 samples of HPV16 infection, and the integration of the virus was determined by ratio analysis. The results showed that there was no correlation between 45 cases (61.64%) as free state, 18 cases (24.66%) as mixed state, 10 cases (13.70%) as integrated state. HPV integration event and clinical stage of cervical cancer (x20.006p0.05) and cervical carcinoma histological type (x20.623p0.05). T178G-A647G-T846C mutation was also not correlated. The presence status of the virus was not related (x2 + 3.696C P 0.05). By analyzing the splicing changes of E6 gene, it was found that the splicing isomer of E6 was mainly E6TI (P0.0001), while the E6 and E6 were not detected in some samples, and the contents of E6 and E6NI were positively correlated (rr 0.6335N P0.001). The integrative events of the virus (FN0.0667P05) and the co-mutation of T178G-A647G-T846C (P0.05) did not affect the changes of E6 splicing. There was no correlation between the changes of E6 splicing and the clinical stage of cervical cancer (F0. 6399P 0.05). 4. The relative content of the transcripts of the three genes of E2E2A647G-T846C in these samples was different (F0. 18.38 / P0.001). The transcription level of E6 and E7 was higher than that of E2 (P0.001 / P0.001). There was no significant difference between the transcription level of E6 and E7 (P0.05). The transcription level of E2 gene was higher in the free state than in the mixed state (p0.01), and the free state was higher than that in the integrated state (P0.05). The transcriptional levels of E6 (p0.01) and E7 (p0.001) were both higher in the integrated state than in the mixed state (P0.01-P0.001) and in the free state (P0.05P 0.05). There was no significant difference in the transcription level of E2E6 and E7 genes in different clinical stages of cervical cancer (P0.05). The cooccurrence of T178G-A647G-T846C did not affect the content of E2OE6E7 gene transcripts (P0.05). Conclusion: in the clinical samples of cervical cancer infected with HPV16 in this study, the detection and analysis of the three cancer-related genes E2E6 and E7 of HPV16 were carried out, although there were differences or correlations between these genes in some aspects. However, no factors related to clinical staging of cervical cancer were found.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【参考文献】