本文選題：HPV16 + E2基因　； 參考：《北京協(xié)和醫(yī)學(xué)院》2014年碩士論文

【摘要】：本研究選取了HPV16型感染的宮頸癌病例樣本(部分含癌旁組織),對(duì)E6、E7基因進(jìn)行測(cè)序分析,分別構(gòu)建進(jìn)化樹,用實(shí)時(shí)定量PCR的方法檢測(cè)DNA水平上E2/E6值以判斷病毒基因組的存在狀態(tài)及E2、E6、E7mRNA水平,并分析E6mRNA的剪接情況,研究結(jié)果如下： 1.E6和E7的建進(jìn)化樹結(jié)果顯示只有亞洲變異型和歐洲變異型沒(méi)有發(fā)現(xiàn)非洲1型、非洲2型、亞-美型和北美變異型。檢測(cè)到T178G-A647G-T846C共突變,其與宮頸癌臨床分期沒(méi)有相關(guān)性(x2=0.046,P0.05)。 2.對(duì)73例HPV16型病毒感染的樣本進(jìn)行E2和E6基因DNA水平的檢測(cè),用比值分析來(lái)判定病毒的整合情況。結(jié)果顯示45例(61.64%)為游離狀態(tài),18例(24.66%)為混合狀態(tài),10例(13.70%)為整合狀態(tài)。HPV整合事件與宮頸癌臨床分期(x2=0.006,P0.05)和宮頸癌組織學(xué)類型(x2=0.623,P0.05)等臨床資料之間沒(méi)有相關(guān)性。T178G-A647G-T846C共突變也與病毒的存在狀態(tài)不相關(guān)(x2=3.696,P0.05)。 3.分析E6基因的剪接變化發(fā)現(xiàn),這些樣本中E6的剪接異構(gòu)體主要以E6*I為主(P0.0001),部分樣本檢測(cè)不到E6*I和E6,且E6和E6*I的含量是呈正相關(guān)的(r=0.6335,P0.001)。病毒的整合事件(F=0.0667,P0.05)和T178G-A647G-T846C共突變(P0.05)均不影響E6剪接變化。E6剪接變化與宮頸癌的臨床分期沒(méi)有相關(guān)性(F=0.6399,P0.05)。 4.E2、E6、E7三個(gè)基因的轉(zhuǎn)錄本在這些樣本中的相對(duì)含量不同(F=18.38,P0.001)：E6和E7的轉(zhuǎn)錄水平均大于E2的轉(zhuǎn)錄水平(P0.001,P0.001),E6和E7的轉(zhuǎn)錄水平間無(wú)顯著性差異(P0.05)。E2基因的轉(zhuǎn)錄水平在病毒游離狀態(tài)高于混合狀態(tài)(p0.01),游離狀態(tài)高于整合狀態(tài)(P0.05)；E6(p0.01)和E7基因(p0.001)的轉(zhuǎn)錄水平都是在病毒整合狀態(tài)高于混合狀態(tài)(P0.01,P0.001)和游離狀態(tài)(P0.05,P0.05)。在宮頸癌不同的臨床分期中,E2、E6和E7基因的轉(zhuǎn)錄水平均無(wú)顯著性差異(P0.05)。T178G-A647G-T846C共突 變不影響E2、E6、E7基因轉(zhuǎn)錄本的含量(P0.05)。 結(jié)論：在本研究涉及的HPV16感染的宮頸癌臨床樣本范圍內(nèi),圍繞HPV16型的E2、E6和E7三個(gè)癌癥相關(guān)基因所進(jìn)行的有關(guān)檢測(cè)和分析中,雖然發(fā)現(xiàn)這些基因在某些方面存在差異或相關(guān)性,但是沒(méi)有發(fā)現(xiàn)與宮頸癌臨床分期相關(guān)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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 趙艷琴;;宮頸脫落細(xì)胞學(xué)和HPV檢測(cè)應(yīng)用于宮頸癌診斷的價(jià)值分析[J];局解手術(shù)學(xué)雜志;2011年06期

2 鄒新慧;葛頌;;基因樹沖突與系統(tǒng)發(fā)育基因組學(xué)研究[J];植物分類學(xué)報(bào);2008年06期

3 陳瑾歆;陳建業(yè);李云祥;;內(nèi)參照基因在實(shí)時(shí)熒光定量RT-PCR檢測(cè)中的應(yīng)用[J];中國(guó)老年學(xué)雜志;2011年14期

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