宫颈脱落细胞hTERC基因的表达及其与宫颈上皮内瘤变的关系
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摘要
在世界范围内,子宫颈癌(Cervical Cancer)是严重威胁妇女健康的主要恶性肿瘤之一。发病率居女性恶性肿瘤的第二位,近年来宫颈癌的发病率逐步上升且趋于年轻化,早期筛查宫颈癌前病变可阻断宫颈癌的发生,目前宫颈癌筛查的主要手段是细胞病理学检查和高危型人乳头状瘤病毒(high risk human papillomavirus,HR-HPV)检测。细胞病理学检查敏感性一般在55%～80%左右,但一旦发现异常细胞,特异性可达到90%以上。HPV检测虽敏感性高,但特异性一般在30%左右,这是由于大多数HPV阳性的病人可自然转阴,并不一定进展为宫颈癌。因而,从宫颈癌筛查及早期诊断角度出发,迫切需要其它可靠指标或手段来协助以上检查,使筛查更准确、可靠及更有预测性。
宫颈上皮内瘤变(cervical intraepithelial neoasia,CIN)发展到宫颈浸润癌需要经过一个较长的阶段,并且低级别宫颈上皮内瘤变绝大多数都可自然回退,仅极少部分进展为癌。近年来的研究表明,人类大多数肿瘤细胞,特别是实体肿瘤细胞,常表现为染色体不稳定性(chromosome instability,CI),即染色体数目的改变和染色体结构异常。宫颈细胞由非典型性发育异常向宫颈癌转变的过程中几乎都伴有3号染色体长臂扩增。其中,涉及到的最重要的基因可能是人类染色体端粒酶RNA亚单位(Humantelomerase RNA component,hTERC)基因。该基因的扩增可阻止细胞凋亡,导致肿瘤产生。因此,hTERC基因的异常扩增可能是宫颈癌形成的早期事件。本研究利用荧光原位杂交(fluorescence in situ hybridization,FISH)方法,通过检测正常宫颈及不同级别上皮内病变患者宫颈脱落细胞内hTERC基因的表达,探讨hTERC基因在子宫颈癌发生、发展进程中所起的作用,为子宫颈癌的筛查及早期诊断提供依据。
材料和方法
1.研究对象
选取2008年6月至2009年1月郑州大学第三附属医院妇科门诊和住院患者共100例,选取2008年3月至2009年3月河南省肿瘤医院妇科门诊和住院患者共79例,收集其宫颈脱落细胞标本。纳入标准:所有研究对象均签署知情同意,同时须有组织病理学结果,按照宫颈上皮内瘤样病变诊断处理规范分为CINⅠ、Ⅱ、Ⅲ级,宫颈癌诊断按照《妇产科学》第六版诊断标准。
排除标准:除外先天性遗传病、妊娠、生殖道急性炎症和严重内科疾病患者。
患者年龄21～61岁,平均36.89±8.72岁。所有患者均行阴道镜检查,并经阴道镜下取活检,常规送病理检查。
2.实验方法
采用液基细胞技术TCT(ThinPrep~(?)Cytology Test)方法采集宫颈脱落细胞,按照2001版的TBS(The Bethesda System)系统进行细胞病理学诊断,分为未见上皮内病变或恶性细胞(Negative for Intraepithelial Lesion or Malignancy,NILM),无明确诊断意义的非典型鳞状上皮细胞(Atypical Squamous Cells of Undetermined Signification,ASC-US)、非典型鳞状上皮细胞不除外高度鳞状上皮内病变(Atypical Squamsuscells-Cannot exclude HSIL,ASC-H)、低度鳞状上皮内病变(Low-grade squamousIntraepithelial Lesion,LSIL)、高度鳞状上皮内病变(High-grade squamous IntraepithelialLesion,HSIL)和宫颈癌标本。利用薄层液基细胞学技术,用其剩余标本5～10ml细胞悬液进行FISH操作,检测不同级别上皮内病变及宫颈癌患者脱落细胞内hTERC基因的表达。采用二代杂交捕获(Hybrid Capture 2,HC2)技术方法检测高危型HPV感染情况。
3.统计学分析
所有数据均采用SPSS11.5进行处理,多样本均数比较采用t检验,率和构成比之间的比较采用x~2检验,以α=0.05作为检验水准。两种诊断方法的比较采用ROC曲线分析。
结果
1.细胞学异常组(ASCUS组、LSIL组、HSIL组、鳞状细胞癌组)与正常细胞学对照组比较,hTERC基因阳性率差异有显著统计学意义(P＜0.001)。
2.HSIL组、宫颈癌组hTERC基因阳性率分别显著高于ASCUS组与LSIL组(P＜0.001)。
3.CINⅡ/Ⅲ组和宫颈癌组的hTE、RC阳性率均显著高于正常对照组,(P＜0.001)。且CINⅡ/Ⅲ组和宫颈癌组的阳性率显著高于CINⅠ组,P值分别为0.0004和0.0001。
4.ASC-US、ASC-H、LSIL、HSIL以上组中高危型HPV DNA阳性率分别为54.8%(17/31)、75.0%(3/4)、60.7%(17/28)和92.3%(12/13),HSIL以上组高危型HPVDNA阳性率明显高于ASC-US组(P＜0.05),ASC-US、ASC-H、LSIL之间相比则差异无显著性(P＞0.05)。
5.以组织病理学诊断为金标准,将CINⅡ及其以上诊断定为阳性,利用ROC曲线比较FISH法检测hTERC基因和细胞学结合TBS分级预测CINⅡ/Ⅲ以上患者的敏感度和特异度。以LSIL为界点,细胞学检测CINⅡ及其以上患者的敏感度为76.8%,特异度为93.7%;以阳性细胞＞8%为界点,FISH法检测CINⅡ及其以上患者的敏感度为79.8%,特异度为83.4%。细胞学和FISH法的ROC曲线下面积分别为:0.913和0.862,二者相比无显著性差异。
结论
1.随细胞学病变程度增加,hTERC基因阳性表达率具有增加趋势。
2.端粒酶hTERC基因的扩增与宫颈癌发生、发展具有相关作用。
3.FISH技术检测上皮内病变和细胞学相比在总体的敏感度和特异度上无显著性差异。但用两种方法作为初筛CIN均有较高价值,两者结合将提高宫颈病变的检出率。
4.在高度上皮内瘤变及宫颈癌中,hTERC基因存在高度表达,hTERC基因检测可作为预测HSIL及宫颈癌的独立指标。
5.高危型HPV感染和宫颈上皮内瘤变关系密切,随着病变程度增加HPV感染率逐渐增高,其检测可以作为宫颈癌筛查的重要指标。
Cervical cancer is one of the most serious threat to women's health worldwide and its incidence is on the second position in female reproductive system cancer. Recent years, the incidence had been found increasing even in younger subjects, thus early screening becomes significantly important. Currently, main methods for early screening depend on cytopathology or detecting the expression of high-risk human papillomavirus (HR-HPV). The sensitivity of cytopathology ranges from 55% to 80%, and its specificity can reached to 90% once abnormal cells are detected. Sensitivity of later is higher, but its specificity is about 30%. As majority of low degreed cervical intraepithelial neopasia (CIN) can return to normal and only small amount of cases process to cancer. The transition from CIN to cervical carcinoma (CC) takes a long time, it's crucial to seek more dependable and predictable methods for early diagnosis.
Recently, some studies found that many types of cancer cells, especially those solid tumor exhibit chromosome instability (CI), such as the chromosome number or structure disorder. Amplification at chromosome 3q almost can be found in the transition from atypical dysplasia to cervical carcinoma and the most possible candidate gene is human telomerase RNA component (hTERC), because its amplification can interrupt cell apoptosis and cause tumor, which may be the early event of malignancy formation.
In our study, we use fluorescence in situ hybridization (FISH) to detect the expression of hTERC in cast-off cells and squamous intraepithelial lesion at all grades and Hybrid Capture 2 (HC2) technology to detect high-risk HPV DNA in order to found possible role of hTERC in the carcinogenesis, development of cervical cancer and found better ways for cervical cancer screening and early diagnosis.
Materials and Methods
1. Subjects
One hundred cytologic specimens of cervix were collected from The Third Affiliated Hospital of Zhengzhou University from June 2008 to January 2009 and 79 cytologic specimens of cervix were collected from Cancer Hospital Henan province from March 2008 to March 2009 according to CIN diagnosis standard. Diagnosis criteria for cervical cancer was based on the textbook "Obstetrics and Gynecology"(sixth edition). The exclusive criteria included pregnancy, genital tract acute inflammation and serious medicine diseases. All patients aged 21-61 (average 36.89±8.72) were checked by colposcopy and biopsy, before the operation after signing consent by the patients.
2. Experimental method
Samples were collected by using liquid-based technology ThinPrep (?) Cytology Test, and the diagnosis standard was based on the "The Bethesda System" (2001 edition) and patients who were diagnosed as Negative for Intraepithelial Lesion or Malignancy (NILM), Atypical Squamous Cells of Undetermined Signification (ASC-US), Atypical Squamsus cells-Cannot exclude HSIL (ASC-H), Low-grade squamous Intraepithelial Lesion (LSIL), High-grade squamous Intraepithelial Lesion (HSIL) and cervical cancer were selected. 5-10ml cell suspension from liquid-based cytology remnants were used to detect hTERC expression of cervical intraepithelial lesion at different grades .
3. Statistical analysis
The differences between research group and control cases were analyzed by the Statistical Packages for the Social Sciences, version 11.5 (SPSS, Chicago). Student's t test for means and reliability analysis was employed. P<0.05 was considered statistically significant.
Results
1. Expression of hTERC at patients (ASCUS, LSIL, HSIL, CC) showed significant difference (P<0. 001) compared with control cases.
2. Expression of hTERC in HSIL and cervical cancer group was significantly higher than ASCUS and LSIL group ( P<0. 001).
3. None of the normal case revealed copy number increase of 3q, but CIN II /III lesions and squamous cervical cancer showed extra copies of 3q. The hTERC gene copy numbers in CIN II /III and cervical cancer group were significantly higher than that in control group (P<0. 001), and significantly higher than CIN I (P>0. 001).
4. Positive rates of high-risk HPV at ASC-US, ASC-H, LSIL, HSIL and SCC were 54.8% (17/31), 75.0% (3/4), 60.7% (17/28) and 92.3% (12/13). positive rate of High-risk HPV DNA at HSIL group is significantly higher than that of ASC-US group (P <0.05), but no difference compared with ASC-H and LSIL respectively (P > 0.05).
5. Using ROC curve method to compare the FISH and cytopathology on patients with CIN II/III, followed by the criterion of pathology diagnosis and CIN II/III as positive case. As for LSIL, the sensitivity of cytopathology was 76.8% and specificity was 93.7%. if taking the cutting edge as the positive cell >8%, the sensitivity of FISH was 79.8% and specificity was 83.4%. The area under ROC curve of the cytopathology and FISH were 0.913 and 0.862 and there was no significant difference between them.
Conclusion
1. With the progress of cervical lesions, the amplification of the hTERC shows an increasing trend.
2. The amplification of the hTERC gene plays an important role in carcinogenesis and development of cervical cancer.
3. Even no significant difference was found, FISH is more sensitive than TCT. Combination of the two methods can improve the diagnosis.
4. Expression of hTERC is increased in high-grade squamous intraepithelial lesion and cervix squamous cells carcinoma and FISH can predict HSIL and cervical cancer independently.
5. High-risk HPV infection is positive related to CIN. Test of High-risk HPV infection can be used as an important indicator for cervical cancer screening.
宫颈上皮内瘤变(cervical intraepithelial neoasia,CIN)发展到宫颈浸润癌需要经过一个较长的阶段,并且低级别宫颈上皮内瘤变绝大多数都可自然回退,仅极少部分进展为癌。近年来的研究表明,人类大多数肿瘤细胞,特别是实体肿瘤细胞,常表现为染色体不稳定性(chromosome instability,CI),即染色体数目的改变和染色体结构异常。宫颈细胞由非典型性发育异常向宫颈癌转变的过程中几乎都伴有3号染色体长臂扩增。其中,涉及到的最重要的基因可能是人类染色体端粒酶RNA亚单位(Humantelomerase RNA component,hTERC)基因。该基因的扩增可阻止细胞凋亡,导致肿瘤产生。因此,hTERC基因的异常扩增可能是宫颈癌形成的早期事件。本研究利用荧光原位杂交(fluorescence in situ hybridization,FISH)方法,通过检测正常宫颈及不同级别上皮内病变患者宫颈脱落细胞内hTERC基因的表达,探讨hTERC基因在子宫颈癌发生、发展进程中所起的作用,为子宫颈癌的筛查及早期诊断提供依据。
材料和方法
1.研究对象
选取2008年6月至2009年1月郑州大学第三附属医院妇科门诊和住院患者共100例,选取2008年3月至2009年3月河南省肿瘤医院妇科门诊和住院患者共79例,收集其宫颈脱落细胞标本。纳入标准:所有研究对象均签署知情同意,同时须有组织病理学结果,按照宫颈上皮内瘤样病变诊断处理规范分为CINⅠ、Ⅱ、Ⅲ级,宫颈癌诊断按照《妇产科学》第六版诊断标准。
排除标准:除外先天性遗传病、妊娠、生殖道急性炎症和严重内科疾病患者。
患者年龄21～61岁,平均36.89±8.72岁。所有患者均行阴道镜检查,并经阴道镜下取活检,常规送病理检查。
2.实验方法
采用液基细胞技术TCT(ThinPrep~(?)Cytology Test)方法采集宫颈脱落细胞,按照2001版的TBS(The Bethesda System)系统进行细胞病理学诊断,分为未见上皮内病变或恶性细胞(Negative for Intraepithelial Lesion or Malignancy,NILM),无明确诊断意义的非典型鳞状上皮细胞(Atypical Squamous Cells of Undetermined Signification,ASC-US)、非典型鳞状上皮细胞不除外高度鳞状上皮内病变(Atypical Squamsuscells-Cannot exclude HSIL,ASC-H)、低度鳞状上皮内病变(Low-grade squamousIntraepithelial Lesion,LSIL)、高度鳞状上皮内病变(High-grade squamous IntraepithelialLesion,HSIL)和宫颈癌标本。利用薄层液基细胞学技术,用其剩余标本5～10ml细胞悬液进行FISH操作,检测不同级别上皮内病变及宫颈癌患者脱落细胞内hTERC基因的表达。采用二代杂交捕获(Hybrid Capture 2,HC2)技术方法检测高危型HPV感染情况。
3.统计学分析
所有数据均采用SPSS11.5进行处理,多样本均数比较采用t检验,率和构成比之间的比较采用x~2检验,以α=0.05作为检验水准。两种诊断方法的比较采用ROC曲线分析。
结果
1.细胞学异常组(ASCUS组、LSIL组、HSIL组、鳞状细胞癌组)与正常细胞学对照组比较,hTERC基因阳性率差异有显著统计学意义(P＜0.001)。
2.HSIL组、宫颈癌组hTERC基因阳性率分别显著高于ASCUS组与LSIL组(P＜0.001)。
3.CINⅡ/Ⅲ组和宫颈癌组的hTE、RC阳性率均显著高于正常对照组,(P＜0.001)。且CINⅡ/Ⅲ组和宫颈癌组的阳性率显著高于CINⅠ组,P值分别为0.0004和0.0001。
4.ASC-US、ASC-H、LSIL、HSIL以上组中高危型HPV DNA阳性率分别为54.8%(17/31)、75.0%(3/4)、60.7%(17/28)和92.3%(12/13),HSIL以上组高危型HPVDNA阳性率明显高于ASC-US组(P＜0.05),ASC-US、ASC-H、LSIL之间相比则差异无显著性(P＞0.05)。
5.以组织病理学诊断为金标准,将CINⅡ及其以上诊断定为阳性,利用ROC曲线比较FISH法检测hTERC基因和细胞学结合TBS分级预测CINⅡ/Ⅲ以上患者的敏感度和特异度。以LSIL为界点,细胞学检测CINⅡ及其以上患者的敏感度为76.8%,特异度为93.7%;以阳性细胞＞8%为界点,FISH法检测CINⅡ及其以上患者的敏感度为79.8%,特异度为83.4%。细胞学和FISH法的ROC曲线下面积分别为:0.913和0.862,二者相比无显著性差异。
结论
1.随细胞学病变程度增加,hTERC基因阳性表达率具有增加趋势。
2.端粒酶hTERC基因的扩增与宫颈癌发生、发展具有相关作用。
3.FISH技术检测上皮内病变和细胞学相比在总体的敏感度和特异度上无显著性差异。但用两种方法作为初筛CIN均有较高价值,两者结合将提高宫颈病变的检出率。
4.在高度上皮内瘤变及宫颈癌中,hTERC基因存在高度表达,hTERC基因检测可作为预测HSIL及宫颈癌的独立指标。
5.高危型HPV感染和宫颈上皮内瘤变关系密切,随着病变程度增加HPV感染率逐渐增高,其检测可以作为宫颈癌筛查的重要指标。
Cervical cancer is one of the most serious threat to women's health worldwide and its incidence is on the second position in female reproductive system cancer. Recent years, the incidence had been found increasing even in younger subjects, thus early screening becomes significantly important. Currently, main methods for early screening depend on cytopathology or detecting the expression of high-risk human papillomavirus (HR-HPV). The sensitivity of cytopathology ranges from 55% to 80%, and its specificity can reached to 90% once abnormal cells are detected. Sensitivity of later is higher, but its specificity is about 30%. As majority of low degreed cervical intraepithelial neopasia (CIN) can return to normal and only small amount of cases process to cancer. The transition from CIN to cervical carcinoma (CC) takes a long time, it's crucial to seek more dependable and predictable methods for early diagnosis.
Recently, some studies found that many types of cancer cells, especially those solid tumor exhibit chromosome instability (CI), such as the chromosome number or structure disorder. Amplification at chromosome 3q almost can be found in the transition from atypical dysplasia to cervical carcinoma and the most possible candidate gene is human telomerase RNA component (hTERC), because its amplification can interrupt cell apoptosis and cause tumor, which may be the early event of malignancy formation.
In our study, we use fluorescence in situ hybridization (FISH) to detect the expression of hTERC in cast-off cells and squamous intraepithelial lesion at all grades and Hybrid Capture 2 (HC2) technology to detect high-risk HPV DNA in order to found possible role of hTERC in the carcinogenesis, development of cervical cancer and found better ways for cervical cancer screening and early diagnosis.
Materials and Methods
1. Subjects
One hundred cytologic specimens of cervix were collected from The Third Affiliated Hospital of Zhengzhou University from June 2008 to January 2009 and 79 cytologic specimens of cervix were collected from Cancer Hospital Henan province from March 2008 to March 2009 according to CIN diagnosis standard. Diagnosis criteria for cervical cancer was based on the textbook "Obstetrics and Gynecology"(sixth edition). The exclusive criteria included pregnancy, genital tract acute inflammation and serious medicine diseases. All patients aged 21-61 (average 36.89±8.72) were checked by colposcopy and biopsy, before the operation after signing consent by the patients.
2. Experimental method
Samples were collected by using liquid-based technology ThinPrep (?) Cytology Test, and the diagnosis standard was based on the "The Bethesda System" (2001 edition) and patients who were diagnosed as Negative for Intraepithelial Lesion or Malignancy (NILM), Atypical Squamous Cells of Undetermined Signification (ASC-US), Atypical Squamsus cells-Cannot exclude HSIL (ASC-H), Low-grade squamous Intraepithelial Lesion (LSIL), High-grade squamous Intraepithelial Lesion (HSIL) and cervical cancer were selected. 5-10ml cell suspension from liquid-based cytology remnants were used to detect hTERC expression of cervical intraepithelial lesion at different grades .
3. Statistical analysis
The differences between research group and control cases were analyzed by the Statistical Packages for the Social Sciences, version 11.5 (SPSS, Chicago). Student's t test for means and reliability analysis was employed. P<0.05 was considered statistically significant.
Results
1. Expression of hTERC at patients (ASCUS, LSIL, HSIL, CC) showed significant difference (P<0. 001) compared with control cases.
2. Expression of hTERC in HSIL and cervical cancer group was significantly higher than ASCUS and LSIL group ( P<0. 001).
3. None of the normal case revealed copy number increase of 3q, but CIN II /III lesions and squamous cervical cancer showed extra copies of 3q. The hTERC gene copy numbers in CIN II /III and cervical cancer group were significantly higher than that in control group (P<0. 001), and significantly higher than CIN I (P>0. 001).
4. Positive rates of high-risk HPV at ASC-US, ASC-H, LSIL, HSIL and SCC were 54.8% (17/31), 75.0% (3/4), 60.7% (17/28) and 92.3% (12/13). positive rate of High-risk HPV DNA at HSIL group is significantly higher than that of ASC-US group (P <0.05), but no difference compared with ASC-H and LSIL respectively (P > 0.05).
5. Using ROC curve method to compare the FISH and cytopathology on patients with CIN II/III, followed by the criterion of pathology diagnosis and CIN II/III as positive case. As for LSIL, the sensitivity of cytopathology was 76.8% and specificity was 93.7%. if taking the cutting edge as the positive cell >8%, the sensitivity of FISH was 79.8% and specificity was 83.4%. The area under ROC curve of the cytopathology and FISH were 0.913 and 0.862 and there was no significant difference between them.
Conclusion
1. With the progress of cervical lesions, the amplification of the hTERC shows an increasing trend.
2. The amplification of the hTERC gene plays an important role in carcinogenesis and development of cervical cancer.
3. Even no significant difference was found, FISH is more sensitive than TCT. Combination of the two methods can improve the diagnosis.
4. Expression of hTERC is increased in high-grade squamous intraepithelial lesion and cervix squamous cells carcinoma and FISH can predict HSIL and cervical cancer independently.
5. High-risk HPV infection is positive related to CIN. Test of High-risk HPV infection can be used as an important indicator for cervical cancer screening.
引文
1.Umayahara K,Numa F,Suehiro Y,et al.Comparative genomic hybridization detects genetic alterations during early stages of cervical cancer progression[J].Genes Chromosomes Cancer,2002,33(1):98-102.
2.Kloth JN,Oosting J.Van Wezel T,et al.Combined array-comparative genomic hybridization and single-nucleotide polymorphism-loss of heterozygosity analysis reveals complex genetic alterations in cervical cancer[J].BMC Genomics,2007,8:53.
3.GBA Wisman,Steven D J,Gert JM,et al.Telomerase in(pre) neoplastic cervical disease[J].Human pathology,2000,31(10):1304-1312.
4.Bauer HM,Ault K.Human papillomavirus:Current prevalence and future protection[J].Sex Transm Dis,2006,33(8):509-11.
5.梁德光,何之恒,蓝柯.人乳头瘤病毒及其致瘤机制的研究进展[J].生命科学,2008,20(6):845-847.
6.Markowitz LE,Dunne EF,Saraiya M,et al.Quadrivalent Human Papillomavirus Vaccine:Recommendations of the Advisory Committee on Immunization Practices (ACIP)[J].MMWR Recomm Rep,2007,56(RR-2):1-24.
7.Wang Q,Griffin H,Southem S,et al.Functional analysis of the human papillomavirus type 16 E1=E4 protein provides a mechanism for in vivo and in vitro keratin filament reorganization.J Virol[3].2004,78(2):821-33.
8.Munoz N,Bosch FX,de Sanjose S,et al.Epidemiologic classification of human papillomavirus types associated with cervical cancer[J].N Engl J Med,2003,348(6):518-27.
9.Feng J,Funk WD,Wang SS,et al.The RNA component of human telomerase[J].Science,1995,269(5228):1236-41.
10.Jeong Seo E,Jung kim H,Jae Lee C,et al.The role of HPV oncoproteins and cellular factors in maintenance of hTERT expression in cervical carcinoma cells[J].Gynecol Oncol,2004,94(1):40-7.
11.Nakano K,Watney E,McDougall JK.Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer[J].Am J Pathol,1998,153(3):857-64.
12.Ohta Y,Yoshiyama H,OhiA.The distinct HPV cervical epithelial cell lines established after transfeetion with HPV16 DNA[J].Jpn J Cancer Res,1999,88(7):654-655.
13.Nair P,Jayaprakash PG,Nir MK,et al.Telomerase,p53 and human papillomavirus infection in the uterine cervix[J].Acta Oncol,2000,39(1):65-70.
14.Snijders PJ,van Duin M,Walboomers JM,et al.Telomerase activity exclusively in cervical carcinomas and a subset of cervical intraepithelial neoplasia grade Ⅲ lesions:strong association with elevated messenger RNA levels of its catalytic subunit and high-risk human papillomavirus DNA[J].Cancer Res,1998,58(17):3812-8.
15.Klingelhutz AJ,Foster SA,McDougall JK.Telomerase activation by the E6 gene product of human papillomavirus type 16[J].Nature,1996,380(6569):79-82.
16.Nagai N,Oshita T,Murakami J,et al.Semiquantitative analysis of telomerase activity in cervical cancer and precancerous lesions[J].Oncol Rep,1999,6(2):325-8.
17.Zhang A,Zheng C,Hou M,et al.Ampliification of the telomerase reverse transcriptase (hTERT) gene in cervical carcinoms[J].Genes Chromosome Cancer,2002,34(3):269-275.
18.张艾芃,李亚里,张全.端粒酶与hTERC基因在宫颈病变中检测及研究[J].四川医学,2008,29(12):1559-1661.
19.Heselmeyer-Haddad K,Janz V,Castle PE,et al.Detection of genomic amplification of the human telomerase gene(TERC) in cytologic specimens as a genetic test for the diagnosis of cervical dysplasia[J].Am J Pathol,2003,163(4):1405-16.
20.余兰,王树玉,贾婵维,等.hTERC基因异常扩增在宫颈病变的临床意义[J].中国优生与遗传杂志,2009,17(4).
21.Olaharskil AJ,Sotelo R,Solorza-Luna G,et al.Tetraploidy and chromosomal instability are early events during cervical carcinogenesis[J].Carcinogenesis,2006.27(2):337-43.
22.Heselmeyer-Haddad K,Sommerfeld K,White NM,et al.Genomic amplification of the human telomerase gene(TERC) in pap smears predicts the development of cervical cancer[J].Am J Pathol,2005,166(4):1229-38.
1.Bauer HM,Ault K.Human papillomavirus:Current prevalence and future protection [J].Sex Transm Dis,2006,33(8):509-11.
2.林晓华,吴宁,侯丽辉,等.人乳头瘤病毒与宫颈肿瘤关系的研究进展[J].医学研究生学报,2006,19(12):183-185.
3.梁德光,何之恒,蓝柯.人乳头瘤病毒及其致瘤机制的研究进展[J].生命科学,2008.20(6):845-847.
4.Markowitz LE,Dunne EF,Saraiya M,et al.Quadrivalent Human Papillomavirus Vaccine:Recommendations of the Advisory Committee on Immunization Practices (ACIP)[J].MMWR Recomm Rep,2007,56(RR-2):1-24.
5.Wang Q,Griffin H,Southern S,et al.Functional analysis of the human papillomavirus type 16 E1=E4 protein provides a mechanism for in vivo and in vitro keratin filament reorganization.J Virol[J].2004,78(2):821-33.
6.Munoz N,Bosch FX,de Sanjose S,et al.Epidemiologic classification of human papillomavirus types associated with cervical cancer[J].N Engl J Med,2003,348(6):518-27.
7.De Filippis RA,Goodwin EC,Wu L,etal.Endogenoushuman papillomavirus E6 and E7 proteins differentially regulate proliferation,senescence,and apoptosis in HeLa cervical carcinomacells[J].J Virol,2003,77(2):1551-1563.
8.Eun Jeong Seo,Hyun Jung Kim,Chan Jae Lee,et al.The role of HPV ncoproteins and cellular factors in maintenance of hTERT expression in cervical carcinoma cells.Gynecol Oncol,2004.94(1):40-47.
9.侯振江,张宗英.端粒酶检测在消化道恶性肿瘤诊疗中的应用[J].国外医学临床生物化学与检验学分册,2002,23(2):109.
10.王建英,范明明.人乳头状瘤病毒感染及端粒酶表达与宫颈癌关系的研究进展[J].医学研究生学报,2008,21(12):1322-1324
11.Feng J,Funk WD,Wang SS,et al.The RNA component of human telomerase[J].Science,1995,269(5228):1236-41.
12.Umayahara K,Hirai Y,Sugiyama Y,et al.Genetic alterations during the progression of early cervical neoplasm[J].Proc Amer Assoc Cancer Res,2005,46.
13.Judith N Kloth,Jan Oosting,Tom van Wezel,et al.Combined array-comparative genomic hybridization and single-nucleotide polymorphism-loss of heterozygosity analysis reveals complex genetic alterations in cervical cancer[J].BMC Genomics.2007,8:53
14.张艾芃,李亚里,张全.端粒酶与hTERC基因在宫颈病变中检测及研究[J].四川医学,2008,29(12):1559-1661.
15.Heselmeyer-Haddad K,Janz V,Castle PE,et al.Detection of genomic amplification of the human telomerase gene(TERC) in cytologic specimens as a genetic test for the diagnosis of cervical dysplasia[J].Am J Pathol,2003,163(4):1405-16.
16.Heselmeyer-Haddad K,Sommerfeld K,White NM,et al.Genomic amplification of the human telomerase gene(TERC) in pap smears predicts the development of cervical cancer[J].Am J Pathol,2005,166(4):1229-38.
17.Andersson S,Wallin KL,Hellstr(o|¨)m AC,et al.Frequent gain of the human telomerase gene TERC at 3q26 in cervical adenocarcinomas[J].British Journal of Cancer 2006,95(3):331-338
18.Nakano K,Watney E,McDougall JK.Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer[J].Am J Pathol,1998,153(3):857-64.
19.Ohta Y,Yoshiyama H,OhiA.The distinet HPV cervical epithelial cell lines established after transfection with HPV16 DNA[J].Jpn J Cancer Res,1999,88(7):654-655.
20.Nair P,Jayaprakash PG,Nir MK,et al.Telomerase p53 and human papillomavirus infection in the uterine cervix[J].Acta Oncol,2000,39(1):65-70.
21.Snijders P J,van Duin M,Walboomers JM,et al.Telomerase activity exclusively in cervical carcinomas and a subset of cervical intraepithelial neoplasia grade Ⅲlesions:strong association with elevated messenger RNA levels of its catalytic subunit and high-risk human papillomavirus DNA[J].Cancer Res,1998,58(17):3812-8.
22.Klingelhutz A J,Foster SA,McDougall JK.Telomerase activation by the E6 gene product of human papillomavirus type 16[J].Nature,1996,380(6569):79-82.
2.Kloth JN,Oosting J.Van Wezel T,et al.Combined array-comparative genomic hybridization and single-nucleotide polymorphism-loss of heterozygosity analysis reveals complex genetic alterations in cervical cancer[J].BMC Genomics,2007,8:53.
3.GBA Wisman,Steven D J,Gert JM,et al.Telomerase in(pre) neoplastic cervical disease[J].Human pathology,2000,31(10):1304-1312.
4.Bauer HM,Ault K.Human papillomavirus:Current prevalence and future protection[J].Sex Transm Dis,2006,33(8):509-11.
5.梁德光,何之恒,蓝柯.人乳头瘤病毒及其致瘤机制的研究进展[J].生命科学,2008,20(6):845-847.
6.Markowitz LE,Dunne EF,Saraiya M,et al.Quadrivalent Human Papillomavirus Vaccine:Recommendations of the Advisory Committee on Immunization Practices (ACIP)[J].MMWR Recomm Rep,2007,56(RR-2):1-24.
7.Wang Q,Griffin H,Southem S,et al.Functional analysis of the human papillomavirus type 16 E1=E4 protein provides a mechanism for in vivo and in vitro keratin filament reorganization.J Virol[3].2004,78(2):821-33.
8.Munoz N,Bosch FX,de Sanjose S,et al.Epidemiologic classification of human papillomavirus types associated with cervical cancer[J].N Engl J Med,2003,348(6):518-27.
9.Feng J,Funk WD,Wang SS,et al.The RNA component of human telomerase[J].Science,1995,269(5228):1236-41.
10.Jeong Seo E,Jung kim H,Jae Lee C,et al.The role of HPV oncoproteins and cellular factors in maintenance of hTERT expression in cervical carcinoma cells[J].Gynecol Oncol,2004,94(1):40-7.
11.Nakano K,Watney E,McDougall JK.Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer[J].Am J Pathol,1998,153(3):857-64.
12.Ohta Y,Yoshiyama H,OhiA.The distinct HPV cervical epithelial cell lines established after transfeetion with HPV16 DNA[J].Jpn J Cancer Res,1999,88(7):654-655.
13.Nair P,Jayaprakash PG,Nir MK,et al.Telomerase,p53 and human papillomavirus infection in the uterine cervix[J].Acta Oncol,2000,39(1):65-70.
14.Snijders PJ,van Duin M,Walboomers JM,et al.Telomerase activity exclusively in cervical carcinomas and a subset of cervical intraepithelial neoplasia grade Ⅲ lesions:strong association with elevated messenger RNA levels of its catalytic subunit and high-risk human papillomavirus DNA[J].Cancer Res,1998,58(17):3812-8.
15.Klingelhutz AJ,Foster SA,McDougall JK.Telomerase activation by the E6 gene product of human papillomavirus type 16[J].Nature,1996,380(6569):79-82.
16.Nagai N,Oshita T,Murakami J,et al.Semiquantitative analysis of telomerase activity in cervical cancer and precancerous lesions[J].Oncol Rep,1999,6(2):325-8.
17.Zhang A,Zheng C,Hou M,et al.Ampliification of the telomerase reverse transcriptase (hTERT) gene in cervical carcinoms[J].Genes Chromosome Cancer,2002,34(3):269-275.
18.张艾芃,李亚里,张全.端粒酶与hTERC基因在宫颈病变中检测及研究[J].四川医学,2008,29(12):1559-1661.
19.Heselmeyer-Haddad K,Janz V,Castle PE,et al.Detection of genomic amplification of the human telomerase gene(TERC) in cytologic specimens as a genetic test for the diagnosis of cervical dysplasia[J].Am J Pathol,2003,163(4):1405-16.
20.余兰,王树玉,贾婵维,等.hTERC基因异常扩增在宫颈病变的临床意义[J].中国优生与遗传杂志,2009,17(4).
21.Olaharskil AJ,Sotelo R,Solorza-Luna G,et al.Tetraploidy and chromosomal instability are early events during cervical carcinogenesis[J].Carcinogenesis,2006.27(2):337-43.
22.Heselmeyer-Haddad K,Sommerfeld K,White NM,et al.Genomic amplification of the human telomerase gene(TERC) in pap smears predicts the development of cervical cancer[J].Am J Pathol,2005,166(4):1229-38.
1.Bauer HM,Ault K.Human papillomavirus:Current prevalence and future protection [J].Sex Transm Dis,2006,33(8):509-11.
2.林晓华,吴宁,侯丽辉,等.人乳头瘤病毒与宫颈肿瘤关系的研究进展[J].医学研究生学报,2006,19(12):183-185.
3.梁德光,何之恒,蓝柯.人乳头瘤病毒及其致瘤机制的研究进展[J].生命科学,2008.20(6):845-847.
4.Markowitz LE,Dunne EF,Saraiya M,et al.Quadrivalent Human Papillomavirus Vaccine:Recommendations of the Advisory Committee on Immunization Practices (ACIP)[J].MMWR Recomm Rep,2007,56(RR-2):1-24.
5.Wang Q,Griffin H,Southern S,et al.Functional analysis of the human papillomavirus type 16 E1=E4 protein provides a mechanism for in vivo and in vitro keratin filament reorganization.J Virol[J].2004,78(2):821-33.
6.Munoz N,Bosch FX,de Sanjose S,et al.Epidemiologic classification of human papillomavirus types associated with cervical cancer[J].N Engl J Med,2003,348(6):518-27.
7.De Filippis RA,Goodwin EC,Wu L,etal.Endogenoushuman papillomavirus E6 and E7 proteins differentially regulate proliferation,senescence,and apoptosis in HeLa cervical carcinomacells[J].J Virol,2003,77(2):1551-1563.
8.Eun Jeong Seo,Hyun Jung Kim,Chan Jae Lee,et al.The role of HPV ncoproteins and cellular factors in maintenance of hTERT expression in cervical carcinoma cells.Gynecol Oncol,2004.94(1):40-47.
9.侯振江,张宗英.端粒酶检测在消化道恶性肿瘤诊疗中的应用[J].国外医学临床生物化学与检验学分册,2002,23(2):109.
10.王建英,范明明.人乳头状瘤病毒感染及端粒酶表达与宫颈癌关系的研究进展[J].医学研究生学报,2008,21(12):1322-1324
11.Feng J,Funk WD,Wang SS,et al.The RNA component of human telomerase[J].Science,1995,269(5228):1236-41.
12.Umayahara K,Hirai Y,Sugiyama Y,et al.Genetic alterations during the progression of early cervical neoplasm[J].Proc Amer Assoc Cancer Res,2005,46.
13.Judith N Kloth,Jan Oosting,Tom van Wezel,et al.Combined array-comparative genomic hybridization and single-nucleotide polymorphism-loss of heterozygosity analysis reveals complex genetic alterations in cervical cancer[J].BMC Genomics.2007,8:53
14.张艾芃,李亚里,张全.端粒酶与hTERC基因在宫颈病变中检测及研究[J].四川医学,2008,29(12):1559-1661.
15.Heselmeyer-Haddad K,Janz V,Castle PE,et al.Detection of genomic amplification of the human telomerase gene(TERC) in cytologic specimens as a genetic test for the diagnosis of cervical dysplasia[J].Am J Pathol,2003,163(4):1405-16.
16.Heselmeyer-Haddad K,Sommerfeld K,White NM,et al.Genomic amplification of the human telomerase gene(TERC) in pap smears predicts the development of cervical cancer[J].Am J Pathol,2005,166(4):1229-38.
17.Andersson S,Wallin KL,Hellstr(o|¨)m AC,et al.Frequent gain of the human telomerase gene TERC at 3q26 in cervical adenocarcinomas[J].British Journal of Cancer 2006,95(3):331-338
18.Nakano K,Watney E,McDougall JK.Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer[J].Am J Pathol,1998,153(3):857-64.
19.Ohta Y,Yoshiyama H,OhiA.The distinet HPV cervical epithelial cell lines established after transfection with HPV16 DNA[J].Jpn J Cancer Res,1999,88(7):654-655.
20.Nair P,Jayaprakash PG,Nir MK,et al.Telomerase p53 and human papillomavirus infection in the uterine cervix[J].Acta Oncol,2000,39(1):65-70.
21.Snijders P J,van Duin M,Walboomers JM,et al.Telomerase activity exclusively in cervical carcinomas and a subset of cervical intraepithelial neoplasia grade Ⅲlesions:strong association with elevated messenger RNA levels of its catalytic subunit and high-risk human papillomavirus DNA[J].Cancer Res,1998,58(17):3812-8.
22.Klingelhutz A J,Foster SA,McDougall JK.Telomerase activation by the E6 gene product of human papillomavirus type 16[J].Nature,1996,380(6569):79-82.