【作者】 余勐；

【导师】 张富春；

【作者基本信息】 新疆大学 ， 动物学， 2006， 硕士

【摘要】 宫颈癌是一类严重威胁妇女生命健康的疾病,宫颈癌的发病率和死亡率都很高,并有逐年上升的趋势。高危型人乳头状瘤病毒(human papilloma- virus,HPV)与宫颈癌发病关系密切,在80-90%的宫颈癌组织中可检测到HPV16和/或HPV18的存在,HPV16和HPV18被认为是妇女宫颈癌的主要病原。流行病学资料表明, HPV感染率很高;高危型HPV存在变异株,其致癌性有改变。新疆南部维吾尔妇女是宫颈癌的高发区,患病率达459～590/10万。其发病与HPV16感染密切相关,宫颈癌组织中HPV16的检出率达77.6-82.6%。目前普遍认为病毒癌蛋白E6、E7在病毒诱导癌症形成过程中起着至关重要的作用,而HPV早期基因的表达受HPV URR(upstream regulatory region,URR)调控,许多转录因子可通过与URR结合调控E6、E7基因的表达,URR突变可改变URR的调控功能,影响病毒癌蛋白E6、E7的表达。为了探讨新疆南部地区维吾尔族妇女宫颈癌组织中人乳头瘤病毒16型上游调控区的变异及其与新疆维吾尔族妇女宫颈癌高发之间的关系,从新疆南部地区维吾尔族妇女宫颈癌组织DNA中PCR扩增HPV16 URR DNA片段,PCR产物直接测序或克隆后测序,分析维吾尔族妇女宫颈癌组织HPV16 URR DNA多态性;然后将URR亚克隆至PGL3-Basic中,转染SiHa和HeLa细胞,48小时后检测荧光素酶活性以评估URR突变造成的转录活性的变化情况。测序结果表明:新疆株URR与原型HPV16 URR比较,新疆株HPV16 URR核苷酸多处发生变异。随后的荧光素酶活性分析证明,突变的新疆株URR转录活性均比德国标准株URR的高,其中新疆株xju-7URR区位于YY1结合位点的7792(C→T)突变和xju-4URR区位于YY1和E2结合区之间7449(T→C)突变,可显著提高启动子的转录活性。成组t检验表明,更多还原

【Abstract】 Cervical cancer is the disease threatened women’s life and health seriously, with the increase of its high suffering and death rate annually. High-risk HPVs (human papillomavirus, HPV) is closely linked to the development of cervical carcinomas. HPV16 and HPV18 are considered to be the major pathogenies, as the HPV16 and/or HPV18 found in tested cervical carcinomatous tissues is 80-90%. The data of cervical cancer epidemiology indicated that the incidence of HPV infection was high, there were variant strains of high-risk HPVs with altered cancer-causing potency. The incidence of cervical cancer was high among Uygur women living in southern Xinjiang and morbidity was 459~590/10,000. The suffering of cervical carcinoma was intimately associated with the infection of HPV16, as HPV16 finding rate in cervical carcinomatous tissues was 77.6- 82.6%. It is generally considered that the virulent oncoproteins E6/E7 plays critical roles in the formation course of HPV-induced cancer. The expression of HPV early gene was regulated by HPV URR, and the expression of HPV E6/E7 genes were regulated by many transcriptional factors via binding to URR. And mutations in HPV URR can alter its regulation function, thus impacting the expression of E6/E7 oncoproteins.In order to investigate the relationship between the mutations in HPV16 type upstream regulatory region(URR) of cervical cancerous biopsies from Uygur women and high incidence of cervical carcinoma among Uygur women in southern Xinjiang, total DNA was extracted from cervical cancerous biopsies. Using these DNA as更多还原