p16、Rb和cyclin D1蛋白在横纹肌肉瘤中的表达及其意义

目的：探讨细胞周期相关蛋白表达异常与横纹肌肉瘤（RMS）的关系。方法：应用S－P免疫组化方法观察p16,Rb和cyclinD1蛋白在RMS中的表达状态。结果：p16,Rb和cyclinD1蛋白表达阳性率分别为55．3％,61．7％和51．1％,在胚胎性横纹肌肉瘤（ERMS）,p16和Rb蛋白表达阳性率为75．0％（21／28）和67．9％（19／28）,明显高于在腺泡状横纹肌肉瘤（ARMS）中的表达（P＜0．05）,p16蛋白阳性率在I,Ⅱ级横纹肌肉瘤分别为75．0％和73．3％,高于Ⅲ级横纹肌肉瘤（45．0％）;p16蛋白阴性及cyclinD1蛋白过表达组,局部浸润,复发或转移发生率高于p16蛋白阳性及cyclinD1蛋白阴性组;11／47例（23．4％）出现两种以上蛋白表达异常。结论：P16,Rb和cyclinD1蛋白异常可能与部分横纹肌肉瘤发生发展有关,且横纹肌肉瘤的发生可能涉及两种以上基因异常。     

人脑胶质瘤中P15蛋白的表达及其临床意义

目的探讨抑癌基因P15蛋白在脑胶质瘤发生、分化过程中的作用及其与预后的关系,以指导临床治疗方案的选择和判断患者的预后。方法采用免疫组化的方法检测68例脑胶质瘤患者病理标本中的P15蛋白,分析P15蛋白的缺失与病理分级及患者预后之间的关系。结果全组共有20例患者(29.4%)标本P15蛋白表达阴性,Ⅲ、Ⅳ级脑胶质瘤的P15蛋白的阴性率明显高于Ⅰ、Ⅱ级脑胶质瘤P15蛋白的阴性率(P<0.05)。P15蛋白表达阳性患者的复发率明显低于P15蛋白表达阴性的患者(P<0.05),分别为25.0%和55.0%。结论P15蛋白的缺失与胶质瘤的恶性程度有关,恶性程度越高,P15蛋白的缺失率越高;P15蛋白的缺失也与患者预后相关,其缺失率越高,患者的复发率也越高,提示P15蛋白缺失的脑胶质瘤患者更易复发,预后也更差。     

胃肠道类癌中p27^kipl、PCNA表达及p16基因缺失

目的：探讨p27^kipl、PCNA及p16与胃肠道类癌转移的关系。方法：采用免疫组化SABC法检测25例胃肠道类癌组织中p27^kipl和PCNA蛋白表达，同时用PCR法检测p16基因的缺失情况。结果：p27^kipl在胃肠道癌阳性表达率为64％（16／25），PCNA I－Ⅱ级和Ⅲ-Ⅳ级表达率分别为56％、44％。2种蛋白阳性表达率与分化程度无相关性（P＞0．05），与淋巴结转移相关（P＜0．05）。p27^kipl与PCNA表达负相关。p16基因纯合性缺失率为48％（12／25），其缺失率与类癌分化程度和淋巴结转移无关。结论：p27^kipl低表达、PCNA高表达与胃肠道类癌转移有关，胃肠道；类癌的发生与p16基因缺失有关，但可能为早期分子事件。     

鼻咽癌组织中p16基因的缺失、高甲基化和蛋白的表达及其临床意义

目的探讨鼻咽癌组织中p16基因的缺失、高甲基化和蛋白的表达及其临床意义。方法应用免疫组织化学EnVision法检测90例鼻咽非角化性癌(NKC)组织p16蛋白的表达缺失,聚合酶链反应和甲基化酶切方法检测23例NKC组织p16基因缺失和高甲基化。结果90例NKC中p16蛋白表达阴性率为46.7%(42/90),而对照组的阴性率为0(0/30),P<0.05。p16蛋白表达与NKC患者的5年生存率有明显的相关性,生存期5年内者,其缺失率为60.0%(36/60);生存期5年以上者,其缺失率为20.0%(6/30),P<0.05。有、无远处器官转移的病例p16蛋白阴性率分别为81.8%(9/11)和41.8%(33/79),P<0.05。而有、无颅底破坏和(或)颅神经侵犯病例p16蛋白阴性率分别为41.7%(10/24)和48.5%(32/66),P>0.05。23例NKC中未检测到p16基因外显子1的缺失,但有10例p16基因外显子2的缺失,缺失率为43.4%(10/23);同时检测到2例外显子1的异常甲基化,高甲基化率为8.7%(2/23);总突变率为52.1%(12/23)。结论在NKC的发生发展过程中,p16基因缺失起着重要的作用。p16蛋白表达与NKC患者的5年生存率和远处转移有一定的相关性,而与NKC的局部组织侵犯无明显相关性。     

原发胃癌中p16和p15基因表达及甲基化异常

为检测胃癌组织中抑癌基因p16,p15及其启动子区甲基化状态和P16、P15蛋白表达情况。选择p16、p15基因及启动子区域，用PCR－SSCP、MSP（甲基化特异的PCR）和测序法对100例胃癌患者的癌组织、癌旁正常组织和5例正常组织进行检测，同时用免疫组化法检测了癌组织和正常对照组织的P16和P15的表达。结果发现癌组织p16和p15基因启动子区甲基化率显著高于癌旁正常组织和正常对照；胃癌组织中，71％的病例P16表达阴性，54％的病例具有p16基因启动子区的高甲基化，无突变和纯合缺失检出；11％的病例P15表达阴性，9％的病例具有p15基因启动子区的高甲基化，p15异常与低分化胃癌有关，p15基因内含子1和外显子1内各发现1例DNA序列改变；癌组织中p16和p15基因启动子区甲基化与其蛋白表达密切相关。结果显示p16基因启动子区域高甲基化是胃癌中p16基因失活的关键因素之一，并在胃癌的发生发展中发挥重要作用；p15基因启动子区域高甲基化在胃癌中起一定作用。     

横纹肌肉瘤p16基因及其蛋白表达的研究

目的探讨ｐ１６基因及其蛋白表达与横纹肌肉瘤发生发展的关系。方法应用ＰＣＲＳＳＣＰ分析及免疫组化方法检测了４４例人横纹肌肉瘤（ＲＭＳ）中ｐ１６基因状态和４７例ＲＭＳ中ｐ１６蛋白表达情况。结果２例有ｐ１６基因纯合性缺失,６例突变。ｐ１６基因异常的８例ＲＭＳ中６例伴广泛浸润、复发或转移。ｐ１６蛋白表达阴性率为３８．３％（１８／４７例）。ｐ１６基因异常的８例中有７例ｐ１６蛋白阴性,１例为阳性。另有１１例ｐ１６蛋白阴性而未查见基因异常。在腺泡状横纹肌肉瘤中,ｐ１６基因异常发生率及ｐ１６蛋白表达缺失率均高于胚胎性横纹肌肉瘤。ｐ１６蛋白阴性的ＲＭＳ更易伴发广泛浸润、复发或转移。结论ｐ１６基因异常及ｐ１６蛋白表达缺失可能与部分横纹肌肉瘤的发生发展有关     

CIC蛋白失表达在少突胶质细胞肿瘤诊断及预后判断中的意义北大核心CSCD

目的探讨CIC蛋白失表达在少突胶质细胞肿瘤中作为预测染色体lp／19q共缺失初筛检测方法的可行性及其对预后的影响。方法回顾性分析首都医科大学宣武医院病理科113例形态学具有少突胶质细胞瘤特征的胶质瘤,分别进行染色体lp／19q共缺失荧光原位杂交（FISH）检测及CIC蛋白免疫组织化学标记。分析CIC蛋白失表达与染色体1p／19q共缺失的相关性,并探讨其与预后的关系。结果在113例组织学诊断为少突胶质细胞肿瘤中,CIC蛋白失表达率为59．3％（67／113）,不同的组织学类型中失表达率不同,在只含有少突胶质细胞成分的肿瘤中失表达率（85．7％．42／49）比混合性的胶质瘤中（39．1％,25／64）高（P〈0．01）。CIC蛋白失表达预测1p／19q共缺失的敏感性和特异性分别为76．1％（54／71）和71．1％（27／38）,假阳性率和假阴性率分别为16．9％（11／65）和38．6％（17／44）。按照2016版（（WHO中枢神经系统肿瘤分类第4版修订版》,本组病例中63例整合诊断符合少突胶质细胞瘤／间变型少突胶质细胞瘤,IDH突变及1p／19q共缺失;CIC蛋白失表达率为81．0％（51／63）,其预测1p／19q共缺失的敏感性和特异性分别提高至81．0％（51／63）和76．9％（20／26）,假阳性率和假阴性率分别降至10．5％（6／57）和37．5％（12／32）。此时,应用Kaplan—Meier法分析CIC蛋白失表达对预后的影响,发现CIC蛋白失表达组比阳性表达组预后稍好,但二者差异无统计学意义（总生存期：P=0．218;无进展生存期：P=0．249）。结论利用免疫组织化学检测CIC蛋白失表达情况,可以作为染色体1p／19q共缺失的一种初筛检测方法。CIC蛋白失表达与预后无关。     

星形细胞瘤中PTEN、p16和p53蛋白表达与预后的关系

目的探讨星形细胞瘤组织中PTEN、p16和p53蛋白表达与患者预后的关系。方法采用EnVision二步法免疫组化技术,检测80例星形细胞瘤组织中PTEN、p16和p53蛋白表达,并结合随访资料进行统计学分析。结果80例中PTEN和p16蛋白的表达缺失分别为17例（21％）和35例（44％）,p53蛋白过度表达为51例（63％）。61例患者获随访结果,随访时间13～47个月,其中29例健在,32例死亡,总生存率为47％;死亡病例中Ⅲ～Ⅳ级患者29例,占9l％。单因素分析示PTEN和p16阳性表达、肿瘤级别低、年龄〈50岁,预示患者预后较好（P〈0．05）;多因素分析结果提示肿瘤级别越高。患者生存期越短（P〈0．001）。结论星形细胞瘤患者的预后与PTEN、p16和p53蛋白的表达有关,PTEN、p16蛋白的丢失预示患者预后较差,3种基因蛋白的表达与肿瘤级别密切相关。     

胃癌P-糖蛋白和p53蛋白协同表达的意义

目的：探讨P糖蛋白和p53蛋白在胃癌中的协同表达意义。方法：应用免疫组化方法检测259例术前未进行化疗的胃癌组织中多药耐药基因产物P 糖蛋白的表达。结果：P－gp和p53在胃癌中的表达分别为26．25％（68／259）和37．01％（96／259）。P－gp表达与胃癌的组织学类型、浸润深度和淋巴结转移状况无关（P＞0．05）;p53除与淋巴结转移状况有关外（P＜0．001）,与组织学类型和浸润深度无关（P＞0．05）。P-gp在p53阳性的病例中的表达明显高于p53阴性的病例,即75％P－gp阳性的病人同时伴有p53阳性。结论：P-gp和p53常协同表达于胃癌组织中,并可作为胃癌病人预后判断和临床耐药的一个可靠指标。     

p16蛋白表达异常与肺癌的关系

目录探讨p16蛋白表达异常与肺癌的关系。方法采用辣根酶标记链霉亲和素免疫组化方法,对35例肺癌组织及相应的癌旁正常肺组织标本和10例肺良性疾病肺组织标本进行研究。结果癌周正常肺组织、肺良性疾病肺组织p16蛋白阳性率均显著高于肺癌组织;临床I,II期p16蛋白阳性率显著高于临床Ⅲ、Ⅳ期;高、中、低分化肺癌细胞p16蛋白表达无显著性差异,男、女肺癌患者p16蛋白表达无显著性差异。结论肺癌的发生可能与p16基因表达产物缺失有关,p16基因的缺失在肺癌进展中起着一定的作用,有可能同时成为预测病程进展的指标之一。     

Uterine leiomyosarcoma arising in leiomyoma: Clinicopathological study of four cases and literature review

In this study we examined four cases of leiomyosarcoma (LMS) arising in leiomyoma (LM). The patients ranged from 40 to 64 years old. Symptoms were unspecific and preoperative diagnosis of LMS had not been made. Size of the tumors ranged from 4 cm to 20.5 cm. All patients were in stage I and no patients developed recurrent disease. Histologically, the LMS component showed moderate to severe cytological atypia and an increased mitotic count in all cases. Tumor cell necrosis in the LMS was seen in three cases. Associated LM component was cellular in one case. The LMS component showed p53 expression in three of four cases and p16 expression was seen in two cases. The LM component lacked p53 expression in all cases and demonstrated p16 expression in two cases, including one case of cellular LM. In all cases, Ki‐67‐positive cells were less than 5% in the LM component and more than 25–30% in the LMS component. This study reveals that: (i) malignant transformation can occur even in relatively small LM; (ii) patients with LMS with an LM component have a favorable prognosis; and (iii) immunohistochemical examination of p53, p16 and Ki‐67 is useful for identification of malignant focus.     

软组织平滑肌肉瘤中p16,细胞周期蛋白D1和pRb蛋白的表达及

目的 探讨软组织平滑肌肉瘤中ｐ１６、细胞周期蛋白Ｄ１和ｐＲｂ的蛋白表达、相互关系及其意义。方法 应用免疫组化方法检测４２例ＬＭＳ患者组织中ｐ１６、ｃｙｃｌｉｎ Ｄ１和ｐＲｂ的蛋白表达情况。结果 ４２例中ｐ１６、ｃｙｃｌｉｎ Ｄ１、ｐＲｂ的总异常率为９７．６％,其中ｐ１６和ｐＲｂ蛋白表达缺失分别为３５．７％和４７．６％,ｃｙｃｌｉｎ Ｄ１过表达５７．１％。ｃｙｃｌｉｎ Ｄ１在平滑肌瘤中也有５５％呈过     

Mechanisms of inactivation of the p16INK4a gene in leiomyosarcoma of soft tissue: decreased p16 expression correlates with promoter methylation and poor prognosis

The p16INK4a tumour suppressor gene, encoding p16 protein, plays a crucial role in regulation of the G1 cell-cycle phase. To investigate the potential role of p16 in soft tissue leiomyosarcoma (LMS), an immunohistochemical analysis was performed of 77 LMSs for p16 expression. Decreased expression of the p16 protein was identified in 25 of 77 LMSs (32%). Decreased expression of p16 correlated significantly with large tumour size (p=0.0038). In a univariate analysis, large tumour size and decreased expression of p16 were statistically significant adverse prognostic factors (p=0.025 and p=0.0021, respectively). In a multivariate analysis including conventional clinicopathological parameters, decreased expression of p16 protein was revealed as the only independent unfavourable prognostic factor (p=0.012). To elucidate the mechanisms of inactivation of the p16INK4a gene, 49 LMSs for which genomic DNA was available were examined; analysis for homozygous deletion, mutation, and promoter hypermethylation was conducted using differential PCR, PCR-SSCP, and methylation-specific PCR, respectively. Promoter hypermethylation was detected in 11 of 49 LMS cases (22%); homozygous deletion was detected in 3 of 49 cases (6%); and mutation was not recognized in any of the cases studied. Eight of 15 cases (53%) with decreased expression of p16 protein revealed methylation of the p16INK4a gene promoter. Promoter hypermethylation correlated closely with decreased expression and poor prognosis (p=0.0014 and p=0.0088, respectively). These results suggest that decreased expression of p16 protein can be considered as an independent reliable prognostic parameter in patients with soft tissue LMS. Furthermore, promoter methylation was more frequent than either homozygous deletion or mutation in this tumour, and promoter methylation was also shown to have a strong association with inactivation of the p16INK4a gene.     

Immunofluorescence analysis of DNA damage response protein p53-binding protein 1 in a case of uterine dedifferentiated leiomyosarcoma arising from leiomyoma

AimsGenomic instability has been indicated during the dedifferentiation process from leiomyoma (LM) to leiomyosarcoma (LMS). Previously, we have described that nuclear expression pattern of DNA damage response protein p53-binding protein 1 (53BP1), detected by immunofluorescence, reflects the magnitude of genomic instability during malignancy. Here, we present a case of LMS arising from LM with molecular analysis of 53BP1, which showed transitional magnitude of DNA damage response within a tumor.Methods and resultsA fifty-year-old female with abdominal mass underwent hysterectomy. Histologically, the tumor consisted of LMS with highly atypical multinucleated giant cells as well as an LM component with transitional atypical spindle cells in the border area. LMS showed diffuse nuclear staining of 53BP1 expression, which has been previously described as high DNA damage response pattern. In contrast, the LM component lacked 53BP1 immunoreactivity and focal expression was observed in transitional lesion. Furthermore, double-labelled immunofluorescence revealed co-localization of 53BP1 with p53 and Ki-67 in the LMS component, which indicated abnormal DNA damage response in proliferative state.ConclusionsThis study revealed that diffuse-type 53BP1 expression may be beneficial to estimate genomic instability during dedifferentiation from LM to DLMS.     

软组织平滑肌肉瘤中p16基因的甲基化检测

目的 探讨软组织平滑肌肉瘤（ＬＭＳ）中p16基因ＩＮＫ４Ａ的甲基化状态及其与p16表达的关系。方法 应用ＭＳＰ法检测３８例软组织平滑肌肉瘤,１０例平滑肌瘤及５例正常平滑肌组织中p16基因ＩＮＫ４Ａ的甲基化状态,用免疫组织化学ＳＰ方法检测p16蛋白表达情况。结果 ３８例ＬＭＳ中９例发生异常甲基化,异常甲基化率为２３.7%（９／３８）。其中,７例p16蛋白表达阴性,２例p16蛋白弱阳性,在p16蛋白表达阴性的ＬＭＳ中,异常甲基化率为５０％（７／１４）。结论 p16基因第一外显子启动子区５‘CpG岛的异常甲基化是导致p16基因失活、蛋白缺如的重要基因外机制,并可能参与肿瘤的发生。     

Clinical significance of p16 protein expression loss and aberrant p53 protein expression in pancreatic cancer

Pancreatic cancer is a disease with poor prognosis mainly due to low resection rates and late diagnosis. To increase resectability and improve survival rates, a better understanding of pancreatic cancer pathogenesis and more effective screening techniques are required. New methods, such as genetic and molecular alterations, may suggest novel approaches for pancreatic cancer diagnosis and treatment. We immunohistochemically investigated 44 formalin-fixed, paraffin-embedded specimens of pancreatic ductal adenocarcinoma using monoclonal anti-p16 antibodies and monoclonal anti-p53 antibodies. The expressions of p16 and p53 proteins were compared using the Chi-square test with SPSS. Disease-free survival was analyzed using the Kaplan-Meier method, verified by the Log- Rank test. Loss of p16 expression was noted in 20 (45.5%) cases and aberrant p53 protein expression was detected in 14 (31.8%) cases. Loss of p16 expression was associated with a higher incidence of lymph node metastasis (p=0.040) and a more advanced stage (p=0.015), although there was no significant correlation between p16 expression and survival. Aberrant p53 protein expression correlated with histologic grade (p= 0.038). Disease-free survival rate was significantly lower in the aberrant p53 protein positive group compared to the negative group (p=0.029). From our results, we suggest that p53 is not a prognostic factor; however, p16 and p53 genes do play important roles in the progression of pancreatic ductal adenocarcinoma.     

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.     

细胞周期抑制蛋白p16、p27蛋白表达与垂体腺瘤侵袭性的关系

目的探讨p27、p16、PCNA的蛋白表达与垂体腺瘤侵袭性的关系,为根治垂体腺瘤及减少肿瘤术后复发提供实验依据。方法应用免疫组化SP法检测43例侵袭性垂体腺瘤和37例非侵袭性腺瘤的p16、p27和PCNA蛋白表达水平,分析p16、p27蛋白表达水平与PCNA蛋白表达之间的相关性。结果垂体侵袭腺瘤p16、p27蛋白表达较非侵袭组明显减低;复发组p16蛋白表达的阳性率与非复发组之间无统计学意义。p27蛋白表达的阳性率较非复发组减低有显著差异。结论p16、p27蛋白表达异常与垂体腺瘤的发生及侵袭性有关,它们的表达情况能作为垂体腺瘤侵袭性的参考指标。     

p73基因在卵巢上皮性肿瘤中的表达及甲基化研究

目的 探讨卵巢上皮性肿瘤中p73蛋白的表达和基因启动子的甲基化情况,并观察其与临床病理学特征的关系.方法 制备包括68例卵巢癌、37例卵巢交界性肿瘤和21例卵巢良性肿瘤的组织芯片,用免疫组织化学EnVision法检测上述组织中p73蛋白表达情况,用亚硫酸氧盐修饰后测序法检测13例新鲜卵巢癌组织及5例新鲜卵巢交界性肿瘤组织的p73基因启动子甲基化情况.结果 92.6％ (63/68)的卵巢癌表达p73,p73蛋白总体表达率均值为32％(p73表达率指p73阳性细胞数所占的百分比),其中浆液性癌( 26/26)的表达率均值为40％,高于其他组织类型的癌(P=0.006).按照卵巢癌发病模式区分,Ⅱ型卵巢癌p73表达率均值(40％)高于Ⅰ型卵巢癌(24％),P=0.010.卵巢癌中p73的表达与临床分期及组织学分级无相关性(均P＞0.05).卵巢交界性肿瘤组(30/37)和良性肿瘤组(12/21)p73的总体表达率均值分别为16％和15％,该两组肿瘤中浆液性肿瘤表达率均值均高于黏液性肿瘤(P-0.003,P=0.026).卵巢癌组的p73阳性表达率均值明显高于交界性肿瘤组和良性肿瘤组(均P ＜0.05),交界性肿瘤组与良性肿瘤组比较差异无统计学意义(P＞0.05).浆液性肿瘤( 49/53)中,卵巢癌组(26/26) p73阳性表达率均值明显高于交界性肿瘤组(12/14)和良性肿瘤组(11/13;P =0.024和P=0.002),而卵巢交界性肿瘤组和良性肿瘤组比较差异无统计学意义(P=0.428).黏液性肿瘤(15/27)中,卵巢癌组(6/7)p73阳性表达率均值高于良性肿瘤组( 1/8;p=0.032),而卵巢癌组与卵巢交界性肿瘤组(8/12)、交界性肿瘤组与良性肿瘤组比较,差异均无统计学意义(P=0.234和P=0.201).p73启动子的甲基化结果显示,13例卵巢癌有8例发生甲基化,但每例样本甲基化频率有所不同,总体甲基化频率均值为8.0％.5例交界性肿瘤有2例发生甲基化,总体甲基化频率均值为9.0％,两组比较差异无统计学意义(P＞0.05).卵巢癌组p73甲基化额率与组织类型、发病模式、组织学分级及临床分期均无相关性(均P＞0.05).结论 卵巢上皮性肿瘤多数表达p73,卵巢癌p73的表达率均值明显高于交界性肿瘤和良性肿瘤,浆液性肿瘤高于其他组织类型;p73蛋白表达率与p73基因甲基化程度不存在简单线性相关关系.