HIF-1α和VEGF在宫颈癌中的表达及相关性研究

目的：探讨缺氧诱导因子-1α（HIF-1α）及血管内皮生长因子（VEGF）在宫颈癌发生发展过程中蛋白表达情况及两者的相关性。方法：应用免疫组织化学S-P方法检测10例正常宫颈（NCE）、18例宫颈上皮内瘤变（CIN）、77例宫颈癌（ICC）组织中HIF-1α和VEGF蛋白的表达情况。结果：在正常宫颈上皮、宫颈上皮内瘤变组织、宫颈癌（Ⅰ—ⅡA期）组织中，HIF．1d蛋白的阳性表达率分别是0．00％、33．33％、70．13％（P〈0．05），VEGF蛋白的阳性表达率分别为10．00％、44．44％、74．00％（P〈0．05）；且随HIF—1α表达增强，VEGF阳性表达率递增，两者呈正相关（P〈0．05）。结论：HIF-1α及VEGF的表达与宫颈癌的发生发展密切相关且两者呈正相关，HIF-1α蛋白可能以转录激活的形式上调VEGF基因的表达，诱导血管生成，促进了宫颈癌的发生发展。     

Epidemiology of cervical cancer in Colombia

Worldwide, cervical cancer is the third most common cancer in women, and the first or second most common in developing countries. Cervical cancer remains in Colombia the first cause of cancer mortality and the second cause of cancer incidence among women, despite the existence of screening programs during the last 3 decades. Bucaramanga, Manizales and Cali reported rates around 20 per 100,000and Pasto 27 per 100,000. The Cali cancer registry has reported a progressive decrease in the age standardized incidence and mortality rates of cervical cancer over the past 40 years. Reasons for the decline in incidence and mortality of cervical cancer are multiple and probably include: improvement in socio-economic conditions, decrease in parity rates and some effect of screening programs. Human papilloma Virus is the main cause of cervical cancer, HPV natural history studies have now revealed that HPVs are the commonest of the sexually transmitted infections in most populations. Most HPV exposures result in spontaneous clearance without clinical manifestations and only a small fraction of the infected persons, known as chronic or persistent carriers, will retain the virus and progress to precancerous and cancer. HPV 16 and 18 account for 70% of cervical cancer and the 8 most common types. (HPV 16, 18, 45, 33, 31, 52, 58 and 35) account for about 90% of cervical cancer. Case-control studies also allowed the identification of the following cofactors that acting together with HPV increase the risk of progression from HPV persistent infection to cervical cancer: tobacco, high parity, long term use of oral contraceptives and past infections with herpes simplex type 2 and Chlamydia trachomatis. The demonstration that infection with certain types of human papillomavirus (HPV) is not only the main cause but also a necessary cause of cervical cancer has led to great advances in the prevention of this disease on two fronts: (i) Primary prevention by the use of prophylactic HPV vaccines; and (ii) secondary prevention by increasing the accuracy of cervical cancer screening.     

高危型人乳头状瘤病毒和宫颈细胞学联合检测在诊断宫颈病变中的临床价值

目的 探讨高危型人乳头状瘤病毒(hish-risk human papillomavirus,HR-HPV)和宫颈细胞学联合检测在诊断宫颈病变中的临床价值.方法 对2004年10月至2006年12月北京大学第一医院就诊的患者进行HR-HPV检测和宫颈细胞学检查,对一项或两项结果异常者均行阴道镜下宫颈活检,并以宫颈活检结果为金标准,比较HR-HPV检测、宫颈细胞学检查、HR-HPV和宫颈细胞学联合检测对宫颈病变的诊断价值.结果 HR-HPV检测、宫颈细胞学检查及HR-HPV检测联合宫颈细胞学检查对诊断宫颈病变有不同价值.HR-HPV检测筛查CINⅡ、CINⅢ的敏感度、特异度、阳性预测值和阴性预测值分别为94.83%、31.06%、55.22%、87.02%,宫颈细胞学筛查CINⅡ、CINⅢ的敏感度、特异度、阳性预测值和阴性预测值分别为92.10%、31.06%、54.50%、81.43%,HR-HPV和宫颈细胞学联合检测筛查CINⅡ、CINⅢ的敏感度、特异度、阳性预测值和阴性预测值分别为99.65%、18.55%、61.46%、97.62%.结论 采用HR-HPV和宫颈细胞学联合检测可提高宫颈病变的检出率,并可指导临床医生对宫颈病变的治疗.     

人端粒酶RNA基因荧光原位杂交检测在宫颈细胞学检查中的应用

目的 探讨人类染色体端粒酶RNA基因(hTERC)扩增的荧光原位杂交(FISH)检测在宫颈脱落细胞防癌筛查中的意义.方法 收集2008年2-10月上海长海医院门诊就诊146例患者宫颈液基细胞学样本,应用间期双色FISH技术检测hTERC基因的扩增情况,并与细胞学和组织学结果进行对照.结果 杂交成功120例(细胞学阴性20例、细胞学阳性100例).hTERC基因扩增的阳性率与细胞学等级成正相关(r=0.465,P＜0.01),与组织学等级成正相关(r=0.610,P＜0.01),各级的阳性率分别为:炎性病变0(0/13)、宫颈上皮内瘤样病变(CIN)Ⅰ级2 8.6%(6/21)、CINⅡ级11/18、CINⅢ级75.0%(18/24)、鳞状细胞癌91.7%(22/24);FISH方法检出高级别(CINⅡ/Ⅲ级)以上病变的敏感性为77.3%(51/66),特异性为82.4%(28/34).阳性预测值89.5%,阴性预测值65.1%,低级别(CIN Ⅰ级)以下病变的hTERC阳性率与高级别以上病变相比差异有统计学意义(x2=32.550,P＜0.01).结合hTERC高倍扩增(信号数＞4)的出现,检出高级别以上病变的敏感性提高到81.2%.结论 hTERC基因扩增的FISH检测有助于辅助细胞学诊断,提高高危病变的检出率.实验结果判断除参照阈值之外,还应结合hTERC基因的扩增类型,出现高倍扩增亦提示高级别以上病变.

Abstract：

Objective To investigate the value of fluorescence in situ hybridization (FISH)detection of human telomerase RNA component ( hTERC ) gene amplification in screening of cervical lesions.Methods A total of 146 post-thinPrep cytology test (TCT) samples were analyzed using FISH by two-color interphase probe targeting hTERC gene at chromosome 3q26 and the data were compared with the cytological and histological results. Results FISH analysis was successful in 120 cases (20 cases of normal and 100 abnormal cases by TCT). Gene amplification of hTERC by FISH had a positive correlation with the cytological (r = 0. 465, P ＜ 0. 01 ) and histological grade results ( r = 0. 610, P ＜ 0. 01 ). Extra copies of hTERC were seen in 28.6% (6/21) of CIN Ⅰ , 61.1% (11/18) of CIN Ⅱ , 75.0% (18/24) of CIN Ⅲ and 91.7% (22/24) of squamous cell carcinoma, respectively. None (0/13) of the inflammation cases showed hTERC amplification. The sensitivity and specificity for detecting high grade lesions by FISH were 77. 3%(51/66) and 82. 4% (28/34) ;and the positive and negative predictive values were 89. 5% and 65. 1%,respectively. The rate of hTERC gene gain in high grade lesions was significantly higher than that in the low grade lesions( x2 =32. 550,P ＜0. 01 ). Combined with the high copy numbers, the sensitivity for detecting high grade lesions was increased to 81.2%. Conclusions Detection of hTERC gene amplification by FISH improves the screening efficiency of high-risk cervical epithelial lesions. The presence of high copy numbers of hTERC correlates with the presence of high grade cervical dysplasia.     

高危型人乳头状瘤病毒DNA检测方法在宫颈疾病中的临床意义

目的评价高危型人乳头状瘤病毒DNA（酶切信号放大法）（Cervista High-risk humanpilloma virus,CervistaHR-HPV）检测方法在宫颈疾病中的临床意义。方法利用CervistaHR-HPV检测方法对437例人组标本进行高危型HPV检测,检测结果与基因测序结果进行对比,同时评价CervistaHR-HPV检测方法在检测CIN2以上的宫颈病变（CIN2＋CIN3＋Cancer,以下简称CIN2＋）的能力,以及三组A5／A6,A7,A9与CIN2＋的相关性。结果CervistaHR-HPV检测方法与基因测序结果相比总体符合率为88．26％,基因测序阳性率为29．08％,本实验为38．96％（P（0．0001）。该检测方法对CIN2＋灵敏度98．46％,特异度58．49％,阳性预测值29．68％,阴性预测值99．54％。A9和CIN2＋的95％OR值的可信区间为10．086～57．283（P〈0．0001）。结论CervistaHR-HPV与基因测序的检测一致性较好,对于CIN2＋灵敏度与特异性较好．可以应用于宫颈癌的防癌筛查,出现A阳性的患者建议进一步行阴道镜检测确诊是否有CIN2＋。     

Simple mucin-type carbohydrate antigens (Tn, sialosyl-Tn, T and sialosyl-T) and gp 230 mucin-like glycoprotein are candidate markers for neoplastic transformation of the human cervix

Mucins and simple mucin-type carbohydrates are cancer-associated antigens in several human tumors. Expression of Tn, sialosyl-Tn, Thomsen-Friedenreich (T), sialosyl-T and of a recently identified mucin-like glycoprotein (gp230) has not yet been thoroughly investigated in human cervix carcinogenesis. In the present study sections from normal cervix (n=10), CIN III lesions (n=10), and invasive carcinomas (n=47) were evaluated immunohistochemically using monoclonal antibodies. In normal cervix there was: cytoplasmatic expression of Tn in 1 case (10%); membranous expression of STn in 8 cases (80%); no expression of T and cytoplasmatic expression of ST in 1 case (10%); gp 230 was expressed in all cases with a membranous pattern. In CIN III lesions there was cytoplasmatic and membranous expression of Tn in 3 cases (30%) and of STn in 9 cases (90%); T and ST were not expressed; gp 230 was expressed in 5 cases (50%) both in the cytoplasm and at the cell membrane. In invasive carcinomas we observed Tn expression in 30 cases (63.8%) and STn in 31 cases (66%); T antigen was not expressed; expression of both ST and gp 230 in 24 cases (51.1%); all antigens showed membranous and cytoplasmatic staining. Our results show that Tn and ST are good markers of invasive carcinomas of the human cervix. We have also shown that loss of expression of the mucin-like glycoprotein gp 230 is associated with malignant transformation at a preinvasive stage. Received: 16 December 1999 / Accepted: 9 February 2000     

宫颈透明细胞癌中人乳头瘤病毒的检测

目的 探讨宫颈透明细胞癌中高危型HPV感染情况.方法 提取1例37岁宫颈透明细胞癌患者手术切除组织蜡块中的DNA,通过巢式PCR方法检测其中HPV感染情况.结果 该患者肿瘤切除组织高危型HPV18型阳性.结论 利用巢式PCR方法分型检测宫颈透明细胞癌中的高危型HPV型别,其准确性及敏感性均较高.     

High-risk human papillomavirus seems not involved in DES-related and of limited importance in nonDES related clear-cell carcinoma of the cervix

Objective

Over 90% of all cervical adenocarcinoma are caused by a transforming infection with a high-risk type human papillomavirus (hrHPV). Previous studies demonstrated that the association between hrHPV positivity and cervical clear-cell adenocarcinoma (CCAC) varies between 0% and 100%. As approximately 60% of all CCAC are associated with intra-uterine diethylstilbestrol (DES) exposure, we determined in a cohort of both DES-exposed and DES-unexposed women the prevalence of hrHPV infections, and the potential etiological role of hrHPV by additional analysis of p16INK4a and p53 expression.

Methods

Representative slides of 28 women diagnosed with CCAC were tested for hrHPV by two PCR methods (the clinically validated GP5+/6+ PCR and the very sensitive SPF₁₀PCR/LiPA₂₅). Fifteen women were DES-exposed, 10 unexposed and of 3 women DES-exposure was unknown. Twenty-one cases with sufficient material were immuno-histochemically stained for p16INK4a and p53.

Results

Seven tumors, of which four DES-exposed and two unexposed tested positive for hrHPV with GP5+/6+ PCR. Thirteen tumors, of which five DES-exposed and seven unexposed, tested positive with SPF₁₀PCR/LiPA₂₅. In one women with unknown exposure, a CCAC tested positive in both assays. Only three cases, none in DES-exposed women, and all positive with both hrHPV assays, revealed diffuse p16INK4a immuno-staining and weak p53 staining as well, supporting indisputable hrHPV involvement.

Conclusions

Although the prevalence of hrHPV was high, only two DES-unrelated CCAC (25%) and one tumor in a woman with unknown exposure could be attributed to hrHPV.     

宫颈周围支持韧带内交感神经分布的研究

目的:观察交感神经在宫颈支持韧带内的分布。方法:用特异性交感神经标记物酪氨酸羟化酶(tyrosine hydroxylase,TH)抗体对主韧带、宫骶韧带及膀胱宫颈韧带不同位置的切片进行免疫组化染色,观察这些韧带内TH阳性纤维的分布。结果:主韧带平均长度5.0cm,距宫颈4.5cm、4.0cm、3.5cm、3.0cm、2.5cm、2.0cm处未见TH阳性纤维,距宫颈1.5cm处TH阳性纤维位于韧带中部,距宫颈1.0cm、0.5cm处TH阳性纤维分布在韧带尾侧。宫骶韧带平均长度6.5cm,距宫颈6.0cm、5.5cm处TH阳性纤维位于韧带头侧的内侧面;距宫颈5.0cm、4.5cm、4.0cm、3.5cm、3.0cm、2.5cm、2.0cm处,TH阳性纤维集中在韧带中下部的外侧面;距宫颈1.5cm、1.0cm、0.5cm处,TH阳性纤维位于韧带尾侧的外侧面。膀胱宫颈韧带平均长2.3cm,距宫颈2.0cm、1.5cm、1.0cm、0.5cm处TH阳性纤维均位于膀胱宫颈韧带深层,散在分布于膀胱静脉内侧。结论:主韧带内交感神经分布在韧带宫颈侧的中下部;宫骶韧带内交感神经自盆壁侧韧带的头侧内侧面向前、向下、向外走行至宫颈;膀胱宫颈韧带内交感神经在韧带深层由宫颈前外侧至膀胱底壁均走行在膀胱静脉内侧。