子宫颈病变中p16INK4A 蛋白表达及其与HPV16/18感染的关系

目的 探讨p16INK4A 蛋白在子宫颈鳞癌(SCC)和子宫颈上皮内肿瘤(CIN)中的表达及其与HPV16/18感染的关系.方法 用原位杂交法检测HPV16/18在25例子宫颈癌、45例CIN及10例慢性子宫颈炎中的表达,同时用免疫组化EliVision法检测p16INK4A 蛋白的表达.结果 (1)与慢性子宫颈炎相比,CIN Ⅱ级、CIN Ⅲ级、浸润癌HPV16/18杂交信号阳性率显著增高(P<0.01);(2)子宫颈鳞癌组织、CIN Ⅰ级、CIN Ⅱ级、Ⅲ级及慢性子官颈炎标本中p16INK4A 蛋白阳性率分别为100.0%、20.0%、46.7%、100.0%和10.0%;(3)在子宫颈鳞癌及CIN HPV16/18感染的标本中p16INK4A 蛋白表达均是阳性.结论 子宫颈鳞癌的形成与HPV感染、p16INK4A 蛋白过表达是呈正相关关系,p16INK4A蛋白可能作为子宫颈鳞癌及CIN的标志物,对子宫颈癌筛查和预防有重要意义.     

人乳头状瘤病毒阴性的宫颈癌及其癌前病变中p16INK4A蛋白表达和DNA倍体分析

目的探讨人乳头状瘤病毒（HPV）感染阴性的宫颈癌及其癌前病变中p16^INK4A蛋白表达和DNA倍体分析的临床病理学意义。方法应用PCR方法筛查出HPV感染阴性的20例慢性宫颈炎、20例宫颈上皮内瘤变（CIN）、3例宫颈腺上皮内瘤变（CGIN）、38例浸润性鳞状细胞癌（鳞癌）和15例浸润性腺癌作为研究对象。应用免疫组织化学（LSAB）染色方法检测p16^INK4A蛋白在这些病变组织中的表达,并结合流式细胞仪DNA倍体分析探讨HPV阴性的宫颈癌的早期诊断和预后判定。结果p16^INK4A蛋白特异性地表达在CIN和CGIN病变、鳞癌以及腺癌细胞的胞核和胞质中,而在正常鳞状上皮和腺上皮中没有任何阳性表达信号。另外,DNA异倍体在浸润性鳞癌和腺癌中的表达率明显高于CIN病变组（P〈0．01）。在有淋巴结转移的浸润癌中DNA异倍体存在的百分率高于无淋巴结转移组,但尚未发现差异有统计学意义。在8例p16^INK4A表达阴性的浸润性鳞癌中有2例表现为DNA异倍体。结论p16^INK4A蛋白检测可以作为HPV感染阴性的宫颈鳞癌及腺癌的早期诊断指标,结合DNA倍体分析将对宫颈恶性肿瘤的诊断有重要的辅助意义。     

HPV16、18 E6蛋白在乳腺癌组织中的表达

目的：观察ＨＰＶ１６ 、１８ Ｅ６ 蛋白在人乳腺癌中的表达并探讨ＨＰＶ１６、１８ 感染与乳腺癌病因学的关系。方法：应用ＳＰ免疫组化法,检测５３ 例乳腺癌、２０ 例正常乳腺组织中ＨＰＶ１６、１８ Ｅ６ 蛋白的表达。结果：乳腺癌组织中ＨＰＶ１６、１８ Ｅ６ 蛋白的阳性表达率为５６－６％ （３０／５３）,多种组织学类型的乳腺癌组织中有ＨＰＶ１６、１８ 型感染的存在。结论：ＨＰＶ１６、１８ 型感染可能与乳腺癌的病因学密切相关。     

高危型人乳头状瘤病毒DNA检测及p16INK4A蛋白表达在子宫颈腺癌诊断中的意义

目的观察p16^INK4A（简称p16）蛋白在子宫颈腺癌和子宫内膜腺癌中的表达及其和高危型人乳头状瘤病毒DNA（HPVDNA）在这两种腺癌中的鉴别诊断价值。方法用免疫组织化学EliVision法检测30例子宫颈腺癌和10例子宫内膜腺癌中p16蛋白的表达情况;并用原位杂交法检测20例子宫颈腺癌和10例子宫内膜腺癌中HPVDNA。结果在30例子宫颈腺癌中21例p16蛋白阳性（70％）,而10例子宫内膜腺癌中3例为阳性。在子宫颈腺癌中p16蛋白呈弥漫性细胞核和细胞质阳性,在子宫内膜腺癌中呈不规则斑点状阳性表达,而且总体强度较弱。HPv16和18在20例子宫颈腺癌中9例（45％）阳性,即细胞核内观察到点状棕黄色颗粒。10例子宫内膜腺癌均为阴性。结论鉴于p16蛋白的表达模式比HPV DNA的检测敏感性高,且易于操作,故建议将其作为区分宫颈癌和子宫内膜腺癌的辅助指标,特别是在活检或诊刮时应增加p16蛋白和HPV DNA的检测。     

cyclin E、p16ink4、ki67在宫颈脱落细胞中的表达及其与HPV16/18感染的相关性

目的　研究cyclinE、p16ink4、ki6 7在宫颈脱落细胞中的表达水平及其与HPV1 6 1 8感染的相关性,探讨其对宫颈癌高危人群筛查的意义。方法　采用免疫组织化学方法对78例宫颈脱落细胞标本进行cyclinE、p16ink4、ki6 7检测,同时应用多重引物PCR技术检测HPV1 6 1 8。结果　cyclinE、p16ink4、ki6 7在宫颈癌细胞中的表达水平均较鳞状上皮非典型增生(ASCUS)差异有统计学意义(P <0 0 0 5 ) ;各级宫颈癌细胞中HPV1 6 的阳性率均较ASCUS差异有统计学意义(χ2 =2 5 2 7,P <0 0 0 5 ) ,且随着宫颈上皮细胞损伤程度加重阳性率升高,差异也有统计学意义(P <0 0 1)。p16ink4和ki6 7在宫颈癌细胞中的表达水平与HPV1 6 高度相关(rs=1 0 ,P <0 0 5 ) ;而cyclinE的表达与HPV1 6 相关性较小(rs=0 4 ,P <0 0 5 )。HPV1 8阳性例数较少,在各项分析中差异均无统计学意义(χ2 =3 6 8,P >0 0 5 )。结论　宫颈癌细胞中cyclinE、p16ink4及ki6 7的高度表达与HPV1 6 感染有关;它们均可能作为有价值的诊断指标应用于宫颈癌高危人群筛查,且cyclinE对宫颈癌的早期诊断意义更大。     

喉癌中HPV感染和p16表达的检测及临床相关性研究

目的：探讨喉鳞状细胞癌中ＨＰＶ感染和ｐ１６ 表达的情况及临床相关性。方法：用免疫组化和原位杂交的方法对５２ 例喉鳞癌组织中ＨＰＶ 感染和ｐ１６ 的表达情况进行检测。结果：ＨＰＶ感染的阳性率为６１－５４ ％（３２／５２） ，ｐ１６ 的阳性率为４４－２３ ％（２３／５２）。结论：ＨＰＶ 感染与ｐ１６ 的表达有关；ＨＰＶ 感染与临床分期、癌细胞的分化程度、肿瘤的淋巴结转移情况、临床分型无关；ｐ１６ 的表达与癌细胞的分化程度、肿瘤的颈淋巴结转移情况有关，与临床分型、分期无关。     

EB病毒感染和p16INK4a蛋白过表达与胃腺癌的关系

目的 探讨Epstein-Barr Virus(EBV)感染与p16异常表达在胃腺癌发生发展中的作用.方法 应用检测免疫组化法检测EB病毒潜伏感染膜蛋白-1(LMP-1)和p16蛋白在97份胃腺癌组织中表达.结果 97份胃腺癌组织中30例LMP-1蛋白表达阳性,阳性率为30.9%,EBV阳性率与患者性别、浸润深度、淋巴结转移、组织学分型和临床分期之间无明显关系(P>0.05);p16蛋白在胃腺癌组织中过度表达,阳性率为63.91%,p16过度表达与患者性别、肿瘤组织学类型、浸润深度不相关,而与淋巴结转移和临床分期相关;EBV感染与p16阳性表达之间无显著相关性(P>0.05).结论 ①EBV感染在胃腺癌的发生中起一定作用;②胃腺癌组织中p16蛋白过度表达是频发事件,p16是重要的预后指标之一;③EBV感染与p16异常表达在胃腺癌发生中是两个独立因素.     

p16和Rb基因蛋白在肺癌中的表达

目的：研究ｐ１６和Ｒｂ基因蛋白的表达与肺癌临床病理学特征的关系。方法：采用免疫组织化学方法对８９例肺癌进行了ｐ１６和Ｒｂ蛋白的定位观察。结果：肺癌；ｐ１６基因蛋白的总丢失率为４７．２％，且与肺癌的组织学类型，淋巴结转移，临床病理分期有关。Ｒｂ基因蛋白的总丢失率为３１．５％，与组织学类型有关。     

宫颈鳞癌中p16、p21WAF1、Rb、cyclinE蛋白的表达

目的通过观察p16、p21WAF1、Rb、cyclinE 4种细胞周期相关蛋白在宫颈鳞状细胞癌中的表达,探讨它们在宫颈癌的细胞周期调控中的作用.方法采用免疫组化Eli Vision二步法对88例宫颈鳞癌组织,16例宫颈上皮内病变(CIN)组织,15例宫颈炎组织进行p16、p21WAF1、Rb、cyclinE 4种蛋白表达的检测.结果p16、p21WAA1、cyclinE在宫颈癌中的表达高于宫颈炎(P＜0.05);Rb在宫颈癌的表达少于宫颈炎(P＜0.05);Rb与p16在宫颈癌中的表达呈负相关(r=-0.675,P＜0.05).结论宫颈癌细胞周期G1期中,由于Rb蛋白的缺失,使cyclinE表达升高,致使癌细胞增生;同时,p16、p21WAF1蛋白在宫颈癌中的表达增高,并失去抑制作用.     

阴茎癌中p53表达与HPV16、18 DNA的检测

目的：研究阴茎癌中p53表达与HPV DNA感染的关系。方法：应用免疫组化技术LSAB法检测33例阴茎癌p53表达及PCR技术检测HPV16、18DNA。结果：33例阴茎癌中，p53阳性表达率51.5%(17/33)；HPV16 DNA阳性率30.3%(10/33)；HPV18 DNA阳性率3.03%(1/33)。结论：阴茎癌中p53表达表明p53突变在阴茎癌发生中有一定意义。阴茎癌发生与HPV16和18型感染有关。     

肺癌组织p16和Rb基因mRNA表达的双重原位杂交观察

目的：研究Ｐ１６和Ｒｂ基因在ｍＲＮＡ表达及其与肺癌的关系。方法：制备ｐ１６和Ｒｂ基因ｃＤＮＡ探针,采用双重原位杂交的方法,地８９例肺癌和正常肺组织进行了Ｐ１６ 一Ｒｂ基因ｍＲＮＡ表达的定位研究。结果：小细胞肺癌ｐ１６ｍＲＮＡ阳性表达率高达８２．４％,而非小细胞肺癌阳性率为４５．８％,两者差异有显著性。非小细胞肺癌ＲｂｍＲＮＡ阳性表达率为８１．９％,而小细胞肺癌一率仅５．９％,两者差异有高度显著性,     

The presence of high-risk HPV combined with specific p53 and p16INK4a expression patterns points to high-risk HPV as the main causative agent for adenocarcinoma in situ and adenocarcinoma of the cervix

Adenocarcinoma in situ (ACIS) and adenocarcinoma (AdCA) of the cervix are frequently missed in population-based screening programmes. Adding high-risk HPV (hrHPV) testing to cervical cancer screening might improve the detection rate of ACIS and AdCA. Since the exact proportion of AdCAs of the cervix that can be attributed to hrHPV infection is still a matter of debate, a comprehensive study was performed of hrHPV presence in ACIS and AdCA of the cervix. Archival formalin-fixed specimens of indisputable ACIS (n=65) and AdCA (n=77) of the cervix were tested for hrHPV DNA by GP5+/6+ PCR-enzyme immunoassay (EIA) and type-specific E7 PCR for 14 hrHPV types. Further immunostaining for p16INK4A and p53 was performed to assess alternative pathways of carcinogenesis potentially unrelated to HPV. hrHPV DNA was found in all (100%) ACISs and 72 (94%) cervical AdCAs, whereas none of 20 endometrial AdCAs scored hrHPV-positive. HPV 18 was most prevalent and found as single or multiple infection in 68% of ACISs and 55% of cervical AdCAs. Diffuse immunostaining for p16INK4a, a potential marker of hrHPV E7 function, was significantly more frequent in hrHPV-positive cervical AdCAs (19/20; 95%) than in those without hrHPV (1/5; 20%; p<0.001). Immunostaining for p53, pointing to stabilized wild-type or mutant p53 protein, was significantly more frequent in hrHPV cervical AdCAs negative for hrHPV (p=0.01). No difference in p16INK4a and p53 immunostaining was found between hrHPV-negative cervical AdCAs and endometrial AdCAs. Hence, only a minority of cervical AdCAs displayed absence of HPV DNA and immunostaining profiles suggestive of an aetiology independent of HPV. Since all ACISs and nearly all cervical AdCAs were hrHPV-positive, the incorporation of hrHPV testing in cervical cancer screening programmes is likely to decrease markedly the incidence of cervical AdCA.     

乳腺浸润性导管癌中HPV18、HPV16感染的研究

目的 :了解乳腺浸润性导管癌中HPV18、HPV16的感染情况 ,分析其是否是乳腺癌发生的危险因素及与临床病理的相关性。方法 :根据HPV16、HPV18的DNA序列 ,合成相应特异的寡核苷酸片段 ,用加尾标记法制备地高辛标记探针 ,用原位杂交法检测 5 1例乳腺浸润性导管癌、10例相应正常乳腺上皮及 15例良性乳腺病变中HPV18、HPV16的感染 ,并分析其与患者发病年龄、肿块大小及淋巴结转移的相关性。结果 :浸润性导管癌中HPV18或 16的总阳性率达 70 6 % ,其中HPV18与HPV16的阳性率分别为 5 8 8%、4 5 1% ,均明显高于正常乳腺上皮的感染率 (30 0 %、10 0 % ;P <0 0 5 ) ;乳腺良性病变的HPV18、16阳性率分别是 6 0 0 %、6 0 % ,其中HPV18的阳性率亦显著高于正常乳腺上皮 (P <0 0 5 )。结论 :(1)HPV16和18可能是乳腺浸润性导管癌发生的致病因子 ,HPV18尚可能与乳腺良性病变的发生有关。 (2 )HPV的感染与患者年龄、肿块大小及淋巴结转移无相关性。     

p16INK4A overexpression and HPV infection in uterine cervix adenocarcinoma

Human papillomaviruses (HPVs) are causally involved in the genesis of cervical carcinomas and their precursors, and there is a strong relationship between the cyclin-dependant kinase inhibitor p16^INK4A and HPV infection. This study was carried out to assess the correlations between p16^INK4A expression as an early biomarker of the endocervical adenocarcinoma and HPV infection. p16^INK4A expression and HPV typing were performed on 46 samples including 5 normal endocervix, 9 benign lesions of the endocervix, 25 endocervical adenocarcinomas, and 7 endometrioid adenocarcinomas of the uterine corpus. A semiquantification of the p16^INK4A immunostaining was realized (using both the staining intensity and the percentage of positive cells) and was graded from 0 to 15. All of the 25 endocervical adenocarcinomas overexpressed p16^INK4A; the adjacent epithelium and the connective tissue were strictly negative. No p16^INK4A was detected in nine benign endocervical lesions and in five normal endocervix. Few endometrioid adenocarcinomas of the uterine corpus that infiltrate the endocervix exhibited a low immunoreactivity (score 0/15 or 1/15). This pattern of expression is significantly associated with HPV infection (p<10 ⁻³), mainly high-risk HPV types (p=0.02). Our results suggest that p16^INK4A is a putative molecular biomarker that consistently discriminates uterine cervix adenocarcinomas from benign lesions and from endometrioid adenocarcinomas of the uterine corpus .     

p16及HPV分型检测在子宫颈黏液腺癌伴双侧卵巢转移中的诊断价值

目的 探讨联合使用免疫组化标记p16及HPV分型检测在诊断和鉴别诊断子宫颈黏液腺癌伴双侧卵巢转移中的诊断价值.方法 分析3例子宫颈黏液腺癌伴双侧卵巢转移的临床病理学特征,对子宫颈及卵巢的癌灶进行含p16在内的多个免疫组化标志物染色及HPV分型检测.结果 3例患者的子宫颈均有不同程度的肥大、糜烂及肿块,双侧卵巢增大,切面有黏液感伴出血、坏死;镜下可见子宫颈腺癌周围有不典型增生的子宫颈管腺上皮,与腺癌移行过渡,可见癌栓,腺癌几乎侵及子宫颈全层;双侧卵巢纤维增生明显,腺癌穿插其中.颈体交界、子宫内膜、输卵管黏膜、阴道手术切缘等查见癌累及.3例子宫颈及卵巢癌灶的免疫表型基本一致,均强阳性表达p16、CK7和Ki-67,HPV亚型分型检测阳性结果如下.子宫颈/卵巢:例1,16、18/16、18、58;例2,16、18/18;例3,16/18.根据p16的表达及HPV分型检测结果,提示3例均为子宫颈黏液腺癌伴双侧卵巢转移.结论 子宫颈黏液腺癌伴双侧卵巢转移属于晚期子宫颈癌.借助p16的表达及HPV亚型的分型检测结果可诊断及鉴别诊断肿瘤的原发部位,有利于肿瘤的后期治疗.     

p16 Immunoreactivity in unusual types of cervical adenocarcinoma does not reflect human papillomavirus infection

Houghton O, Jamison J, Wilson R, Carson J & McCluggage W G
(2010) Histopathology 57, 342–350
p16 Immunoreactivity in unusual types of cervical adenocarcinoma does not reflect human papillomavirus infection Aims: The association between human papillomavirus (HPV) and cervical carcinoma is well known, with HPV being identifiable in almost all cervical squamous carcinomas and most adenocarcinomas. However, the prevalence of HPV in unusual morphological types of cervical adenocarcinoma has not been investigated extensively. The aim was to determine HPV status in a series of primary cervical adenocarcinomas, enriched for unusual morphological types. The relationship between HPV and p16 immunoreactivity in these neoplasms was also investigated, as it is generally assumed that in cervical neoplasms diffuse p16 expression is predictive of the presence of high‐risk HPV. Methods and results: Sixty‐three cervical adenocarcinomas, comprising those of usual type (n = 43), minimal deviation type (n = 4), gastric type (n = 3), intestinal type (n = 3), mesonephric type (n = 3), clear cell type (n = 4), serous type (n = 2) and hepatoid type (n = 1) underwent linear array HPV genotyping and immunohistochemistry for p16. Overall, HPV was identified in 32 of 56 cases (57%) in which sufficient DNA was present for analysis. The most common HPV types were 16 and 18, with these being identified in 20 and 18 cases, respectively, either alone or in combination. Seventy‐eight per cent of usual‐type adenocarcinomas were HPV‐positive, as was the single serous carcinoma in which there was sufficient DNA for analysis. In contrast, all minimal deviation adenocarcinomas and those of gastric, intestinal, mesonephric and clear cell types were HPV‐negative, as was the single hepatoid carcinoma. All usual‐type adenocarcinomas exhibited p16 immunoreactivity (diffuse staining in all but one case), as did 11 of 20 of those of unusual morphological type (five focal, six diffuse). Conclusions: Most, but not all, cervical adenocarcinomas of usual type contain HPV, but those of unusual morphological type are almost always HPV‐negative. This has implications for the efficacy of HPV vaccination in the prevention of cervical adenocarcinoma. A significant proportion of cervical adenocarcinomas are p16‐positive in the absence of HPV, illustrating that in these neoplasms diffuse p16 immunoreactivity is not a reliable surrogate marker of the presence of high‐risk HPV.     

宫颈正常细胞和宫颈鳞状细胞癌、腺癌组织中HPV感染基因型的分布

目的 观察宫颈正常细胞、宫颈鳞状细胞癌和腺癌组织标本中人乳头瘤病毒(human papillomavirus,HPV)感染的基因型分布情况及临床意义.方法 采用PCR和基因芯片检测技术对1 047例宫颈正常细胞、161例宫颈鳞状细胞癌和82例宫颈腺癌组织标本进行23种HPV基因分型检测,并对受检者进行相关资料分析.结果 1 047例宫颈细胞正常者检出HPV感染为109例,HPV感染率为10.41％(109/1 047);161例宫颈鳞状细胞癌检出HPV感染者146例,HPV感染率为90.68％(146/161);82例宫颈腺癌检出HPV感染者59例,HPV感染率为71.95％(59/82).结论 PCR与基因芯片检测技术可用于宫颈细胞和组织标本检测,一次可检测23种HPV基因型,对我国女性宫颈HPV感染的分子流行病学调查、宫颈鳞状细胞癌及腺癌的防治及其疫苗的研究具有十分重要的意义.