肾细胞癌RASSF1A基因启动子区甲基化状况与表达水平研究

目的检测19例肾细胞癌患者癌组织及癌旁组织Ras相关区域家族蛋白1A（RASSFlA）基因启动子区甲基化情况并检测其mRNA表达水平，探索两者之间的联系。方法收集肾细胞癌患者癌组织及相应癌旁组织19份，分别提取其基因组DNA，以甲基化特异性PCR（Methylation special PCR，MSP）法检测RASSFlA基因启动子区甲基化情况。并以QPCR法检测了RASSFlA基因的mRNA表达水平。结果19例中有lO例肾细胞癌患者癌组织存在高甲基化，mRNA表达水平表达降低，二者之间存在显著负相关性（r=-0．8734，P〈0．01）。结论肾细胞癌中RASSFlA基因启动子区存在高甲基化，并抑制该基因的表达。     

GnRH在宫颈鳞状细胞癌中的表达及意义

目的:从蛋白质及核酸水平研究GnRH(促性腺激素释放激素)在宫颈鳞状细胞癌中的表达,以探讨其存在的临床意义。方法:采用免疫组化和原位杂交两种实验方法,分别从蛋白质和核酸水平检测45例宫颈鳞状细胞癌及20例慢性宫颈炎中GnRH的表达。结果:在20例慢性宫颈炎宫颈上皮组织中GnRH及GnRH mRNA表达均为阴性,而在45例宫颈鳞状细胞癌组织中,GnRH阳性表达率为80%,GnRH mRNA阳性表达率为64.44%,即无论是在蛋白质水平还是在核酸水平比较两者均显示差异有显著性;定性及定量比较GnRH及GnRH mRNA在不同临床期别的宫颈癌组织中的表达情况,结果为GnRH定性表达率Ⅰ期50%(6/12),Ⅱa期85.7%(18/21),Ⅱb-期100%(12/12);GnRH mRNA定性表达率,Ⅰb期33.3%(4/12),Ⅱa期71.4%(15/21),Ⅱb-期83.3%(10/12);GnRH及GnRH mRNA在定性上表达有显著差别(P<0.05)。结论:宫颈鳞状细胞癌组织存在较高的GnRH,其有可能参与宫颈鳞状细胞癌发生、发展,为进一步研究通过抑制GnRH的产生,减少宫颈癌的发生及治疗宫颈癌提供了一定的理论基础。     

Lewis血型相关抗原在宫颈鳞状细胞癌中的表达

Ｌｅｗｉｓ血型相关抗原在宫颈鳞状细胞癌中的表达周先荣童夙明杜心谷Ｌｅｗｉｓ血型及其相关抗原是一类已被阐明结构的糖类抗原，我们用免疫组化ＡＢＣ法检测６种Ｌｅｗｉｓ血型相关抗原在正常宫颈鳞状上皮和鳞状细胞癌中的表达及其改变。一、材料与方法１材料：３９例...     

广东省潮汕地区食管鳞状细胞癌中p53蛋白的表达

采用ＣＭ－１和ＤＯ－７一抗的免疫组化法，检测了广东省潮汕地区５５例食管鳞状细胞癌中的ｐ５３蛋白 表达的现象。ｐ５３蛋白阳性细胞不但可在呈浸润生长的肿瘤细胞中见到，而且在已具有浸润癌巢ｐ５３蛋白表达的肿瘤癌旁浸润前病变如上皮高度增生，异型增生和原位癌中可出现。     

泛素特异性蛋白酶10在宫颈鳞状细胞癌中的表达及临床意义

目的:研究泛素特异性蛋白酶10(ubiquitin-specific protease 10,USP10)蛋白在人体宫颈鳞状细胞癌组织中的表达及临床意义.方法:采用免疫组织化学方法检测105例人体宫颈鳞状细胞癌及65例正常鳞状上皮组织石蜡切片中USP10蛋白的表达水平,并分析其表达与临床病理因素之间的相关性.结果:宫颈鳞状细胞癌组织中USP10阳性表达率为41.9%,而宫颈正常鳞状上皮组织中USP10的阳性表达率为61.5%.与正常鳞状上皮组织相比,USP10蛋白在宫颈鳞状细胞癌组织中的表达明显下调,差异具有统计学意义.同时,USP10蛋白表达与宫颈鳞状细胞癌的FIGO临床分期及肿瘤浸润深度呈负相关,而与E-cadherin,P53蛋白表达呈正相关.结论:宫颈鳞状细胞癌组织中USP10蛋白表达的下调与宫颈鳞状细胞癌的恶性程度及预后相关.     

SNIP1在宫颈鳞状细胞癌中的表达及意义

目的:探究Smad核内相关蛋白1(Smad nuclear interacting protein 1,SNIP1)在正常宫颈上皮组织及宫颈鳞状细胞癌(宫颈鳞癌)组织中的表达及其意义.方法:免疫组织化学方法及实时荧光定量PCR(Real-time PCR)技术检测SNIP1在23例正常宫颈组织(正常宫颈组)、25例早期宫颈鳞癌组织(Ⅰ a～Ⅱa期)(早期宫颈鳞癌组)和45例晚期宫颈鳞癌组织(Ⅱb～Ⅲb期)(晚期宫颈鳞癌组)中的表达.结果:免疫组织化学方法显示SNIP1蛋白在正常宫颈组、早期宫颈鳞癌组及晚期宫颈鳞癌组中胞质强阳性表达率分别为0,36.0％和73.3％;胞核强阳性表达率分别为17.4％,44.0％和71.1％,且三组间胞质与胞核的表达差异均具有统计学意义(P＜0.01).在伴有淋巴结转移的宫颈鳞癌病例中,SNIP1在胞质、胞核中的强阳性率分别高于不伴淋巴结转移病例,且差异有统计学意义(P＜0.001).Real-time PCR技术检测显示SNIP1 mRNA在正常宫颈组、早期宫颈鳞癌组及晚期宫颈鳞癌组中表达呈递增趋势,除了正常宫颈组与早期宫颈鳞癌组间差异无统计学意义外,其余各组间差异有统计学意义(P＜0.05).结论:SNIP1可能参与调控宫颈鳞癌的进展及侵袭转移过程的发生,并有望成为宫颈鳞癌侵袭转移情况的预测因子.     

宫颈微小浸润性鳞状细胞癌

宫颈微小浸润癌（microinvasive cervical cancer,MIC）的概念提出已经有100多年了,对于其病理诊断标准及临床治疗方法一直存在争议,但随着对其研究的广泛深入,特别是对微小浸润性鳞状细胞癌的研究,所涉及的样本量比较大,随访时间充分,得到了一些有意义的研究成果。相比较而言,对于宫颈微小浸润性腺癌的研究还不够全面,我们仅就近十几年来对于宫颈微小浸润鳞状细胞癌的研究及探讨进行概述,以供病理和临床医生参考。     

人喉鳞状细胞癌p53蛋白和PCNA的表达

目的：探讨Ｐ５３蛋白和增殖细胞核抗原（ＰＣＮＡ）在人喉鳞状细胞癌的表达。方法：应用免疫组织化学Ｓ－Ｐ法对８０例喉鳞状细胞癌、１０例声带息肉、１０例喉乳头状瘤进行检测。结果：（１）Ｐ５３蛋白在喉鳞状细胞癌阳性率为４１％，阳性细胞指数为５４．５９±２２．８２，与临床Ｔ分期，分化程度以及淋巴结转移无关；（２）ＰＣＮＡ指数在声带息肉，喉乳头状瘤，喉鳞的逐渐提高，在有淋巴结转移的比无淋巴结转移的指数明显增高     

p16在原发性肺鳞状细胞癌与肺转移性宫颈鳞状细胞癌鉴别诊断中的价值

宫颈癌最常见的组织学类型是鳞状细胞癌(SCC),据报道有3.1%～8.2%宫颈癌可发生肺转移.区分肺原发性SCC与转移性宫颈SCC对评价病人预后和选择治疗措施非常重要.尽管肺转移性癌为多灶性病变,但有时也可表现为孤立性病灶,导致二者的鉴别非常困难,到目前为止还没有可靠的免疫标记物可用于二者的鉴别.     

宫颈鳞状细胞癌组织中TGF-β_1、p53基因的原位杂交检测

宫颈癌是最常见的女性生殖道恶性肿瘤，死亡率居妇科肿瘤第２位，其发生是多因素、多步骤的复杂过程〔１〕。通过研究宫颈鳞状细胞癌组织中ＴＧＦ－β１、ｐ５３基因的变化，进一步揭示宫颈癌发生的分子机制。１材料和方法１．１标本２２例宫颈鳞状细胞癌均为本院妇产科手...     

启动子DNA甲基化调节胃癌中PDX1基因表达

目的明确PDXl启动子DNA甲基化,探讨启动子甲基化对PDXl在胃癌中表达的调节作用。方法收集3例胃癌活检组织,免疫组化检测PDXl蛋白表达：吉西他滨处理3株胃癌细胞,RT—PCR检测不同药物剂量和作用时间下PDXlmRNA表达;构建PDXl报告基因,检测启动子活性及吉西他滨处理前后启动子活性的变化;甲基化特异性PCR（MSP）检测3株胃癌细胞和8对配对胃癌组织中PDXl启动子甲基化状态。结果免疫组化结果显示胃癌中PDXl表达低于正常胃黏膜;RT—PCR显示吉西他滨使PDXlmRNA重获表达,且随剂量和时间依赖性。F383有最强启动子活性,吉西他滨显著增加了PDXl启动子活性（P〈0．05）。F383在AGS、BCG823、SGC7901中呈DNA完全甲基化状态;87．5％的胃癌组织出现F383部分甲基化,12．5％出现完全甲基化。癌旁正常组织仅有37．5％出现F383部分甲基化,未出现完全甲基化,两者比较有显著性差异（P〈0．05）。结论PDXl启动子存在DNA高甲基化,抑制了胃癌中PDXl的表达。     

Mismatch repair in Gram-positive bacteria

DNA mismatch repair (MMR) is responsible for correcting errors formed during DNA replication. DNA polymerase errors include base mismatches and extra helical nucleotides referred to as insertion and deletion loops. In bacteria, MMR increases the fidelity of the chromosomal DNA replication pathway approximately 100-fold. MMR defects in bacteria reduce replication fidelity and have the potential to affect fitness. In mammals, MMR defects are characterized by an increase in mutation rate and by microsatellite instability. In this review, we discuss current advances in understanding how MMR functions in bacteria lacking the MutH and Dam methylase-dependent MMR pathway.     

Aberrant promoter methylation of the PAQR3 gene is associated with prostate cancer

Methylation markers are promising tools for diagnosis, prognosis and targeted treatment of cancer. In prostate carcinoma, aberrant promoter hypermethylation occurs earlier in the disease course and more consistently than recurrent somatic mutations. PAQR3, a tumor suppressor gene, was recently found to be downregulated in prostate cancer cell lines. We hypothesized that promoter methylation could be responsible for PAQR3 silencing in prostate cancer tissues. We aimed to investigate PAQR3 promoter methylation in prostate cancer by comparing it to benign prostatic hyperplasia (BPH). A total of 154 human prostate tissue samples, including 92 cases with prostate cancer and 62 cases with BPH, were examined by methylation-specific PCR. Clinicopathological correlation between PAQR3 promoter methylation and prognostically relevant variables was studied by statistical analysis. Promoter methylation of PAQR3 was significantly more frequent in prostate carcinoma compared to BPH (73.9% vs. 25.8%, p < 0.01). The high prevalence of PAQR3 methylation in cancer foci was also confirmed with microdissection technique in 12 samples of prostate adenocarcinoma. PAQR3 hypermethylation was associated with perineural invasion (p = 0.03), an adverse clinicopathological feature of prostate cancer. We concluded that PAQR3 can be a promising methylation marker candidate for the detection and monitoring of prostate cancer.     

Retinoic acid receptor beta promoter methylation and risk of cervical cancer

Cervical cancer is one of the leading causes of death in women worldwide, particularly in developing countries. Human papillomavirus has been reported as one of the key etiologic factors in cervical carcinoma. Likewise, epigenetic aberrations have ability to regulate cancer pathogenesis and progression. Recent research suggested that methylation has been detected already at precancerous stages, which methylation markers may have significant value in cervical cancer screening. The retinoic acid receptor beta (RARβ) gene, a potential tumor suppressor gene, is usually expressed in normal epithelial tissue. Methylation of CpG islands in the promoter region of the RARβ gene has been found to be associated with the development of cervical cancer. To investigate whether RARβ methylation is a potential biomarker that predicts the progression of invasive cancer, we reviewed 14 previously published articles related to RARβ methylation. The majority of them demonstrated that the frequency of RARβ promoter methylation was significantly correlated with the severity of cervical epithelium abnormalities. However, methylation of a single gene may not represent the best approach for predicting disease prognosis. Analyzing combinations of aberrant methylation of multiple genes may increase the sensitivity, and thus this approach may serve as a better tool for predicting disease prognosis.     

Mismatch repair genes in renal cortical neoplasms

Mutation of human mutL homolog 1 (MLH-1) and human mutS homolog 2 (MSH-2) has been linked with the pathogenesis of colorectal carcinoma in hereditary nonpolyposis colorectal cancer syndrome and other carcinomas. Mutations of these genes in renal cell carcinomas were recently described. The aim of this study was to examine the expression of MLH-1 and MSH-2 in renal cortical neoplasms of various histological types by immunohistochemistry. Thirty-eight (n = 38) resected renal tumors were obtained from the surgical pathology files of the UMass Memorial Healthcare, including clear cell carcinomas (CLEARs, n = 20), papillary carcinomas (PAPs, n = 8), chromophobe carcinomas (CHRs, n = 4), and oncocytomas (ONCs, n = 6). Positive immunostaining for MLH-1 and MSH-2 was graded by the number of positive tumor cell nuclei, as follows: 0, negative; 1, up to one third of positive nuclei; 2, one to two thirds positive; and 3, greater than two thirds positive. Loss of MLH-1 or MSH-2 was defined as a tumor with grade 0 or 1, compared with the normal tubules. Normal tubules and intercalated ducts contained cells positive for MLH-1 and MSH-2 in all cases. For both antibodies, positive staining in tumors ranged from grade 1 to 3 in the CLEAR and PAP but was only grade 2 to 3 in the CHR and ONC. Loss of MLH-1 and/or MSH-2 occurred in malignant tumors but not in ONC. Loss of MLH-1 was present in 8 (40%) of 20 CLEARs and 4 (50%) of 8 PAPs, compared with loss of MSH-2 in 4 (20%) of 20 CLEARs and 1 (25%) of 4 CHRs. Our results suggest that loss of mismatch repair genes is involved in the malignant transformation in some renal carcinomas, particularly those derived from the proximal tubules.     

Fibulin-1基因启动子区域甲基化与食管癌的关系研究

目的:检测食管癌Fibulin-1基因启动子区域甲基化的情况,探讨其相关的临床意义.方法:选取黄石市中心医院2014年1月至2016年2月收治的食管鳞状细胞癌患者96例,另取同期黄石市中心医院因食管良性病变手术切除的食管组织40例作为对照.Fibulin-1基因启动子甲基化使用亚硫酸盐修饰后PCR检测.免疫组织化学检测显示Fibulin-1蛋白表达.结果:食管癌患者中Fibulin-1基因启动子甲基化阳性率为36.5％,显著高于对照组的15.0％(x2=2.517,P=0.036).免疫组织化学检测显示Fibulin-1基因启动子甲基化标本中Fibulin-1蛋白阳性表达率为90.2％.食管癌患者男女性别和不同年龄之间Fibulin-1基因启动子甲基化率无显著性差异(P＞0.05).Ⅲ,Ⅳ临床分期患者Fibulin-1基因启动子甲基化率为43.1％,显著高于Ⅰ,Ⅱ临床分期患者的28.9％(x2=2.426,P=0.046).肿瘤有淋巴结转移患者Fibulin-1基因启动子甲基化率为44.4％,显著高于无淋巴结转移患者的26.2％(x2=2.438,P=0.041).肿瘤中、低分化患者Fibulin-1基因启动子甲基化率为42.6％,显著高于高分化患者的25.7％(x2=2.431,P=0.043).结论:Fibulin-1基因启动子在食管癌组织中高甲基化,Fibulin-1基因启动子高甲基化与食管癌的临床分期、肿瘤分化程度以及有无淋巴结转移密切相关.     

DAPK1, MGMT and RARB promoter methylation as biomarkers for high-grade cervical lesions

Gene promoter methylation may be used a potential biomarker for detecting solid tumor including cervical cancer. Here, we used methylation sensitive-high resolution melting (MS-HRM) analysis to detecting promoter methylation ratios of DAPK1, MGMT and RARB gene in patients with different cervical disease grade. The detection of gene promoter methylation was conducted in two hundred fifty patients’ samples including normal cytology (n=48), cervical intraepithelial neoplasia grade 1 (CIN1, n=54), cervical intraepithelial neoplasia grade 2 (CIN2, n=47), cervical intraepithelial neoplasia grade 3 (CIN3, n=56) and cervical squamous cell carcinomas (SCS, n=45). We found there were a significant positive correlation between the promoter methylation status of DAPK1 and cervical disease grade (P=0.022). In addition, the methylated promoters of DAPK1 combined with MGMT, MGMT combined with RARB, DAPK1 combined with RARB were positive correlated with cervical disease grade (P < 0.05). All three genes promoters methylated were positive correlated with cervical disease grade (P < 0.001). Receiver operating characteristic (ROC) curves was conducted to evaluate whether the three genes methylation could be used to be a potential marker for diagnosing high grade cervical disease (HSIL and SCC). The cutoff values for the methylation rates of all these genes were 0-5%. Regrettably, only the methylation of MGMT combined with DAPK1 gave 43.4% sensitivity and 68.6% specificity. The current results indicated that MS-HRM-based testing for DNA methylations of MGMT plus DAPK1 genes holds some promise for high grade cervical disease screening.     

RUNX3基因启动子甲基化与胃癌关系的meta分析

目的采用meta分析的方法评价RUNX3基因启动子甲基化与胃癌发生的关系。方法检索公开发表在Pubmed、EMBASE、Ovid、中国期刊全文数据库（CNKI）关于RUNX3基因启动子甲基化与胃癌发生关系的研究。应用Statall．0统计软件分析RUNX3基因启动子甲基化与胃癌发生间的关联。结果共有19项研究纳入分析．meta分析结果显示：胃癌患者癌组织中RUNX3基因启动子甲基化发生率为P=55％（95％CI：45％～66％）：胃癌患者远癌正常胃组织中RUNX3基因启动子甲基化发生率为P=18％（95％CI：7％～29％）。与远癌正常胃组织相比,胃癌组织中RUNX3基因启动子甲基化发生优势比OR=7．85（95％CI：5．81～10．61）。结论胃癌患者癌组织中RUNX3基因启动子甲基化发生率明显高于远癌正常胃组织,RUNX3基因启动子甲基化可能与胃癌的发生密切相关。     

肺癌患者PTEN基因启动子高甲基化的检测

目的 探讨肺癌患者组织、外周血浆及支气管肺泡灌洗液(BALF)中张力蛋白同源的磷酸酶基因(PTEN)启动子异常基因化状况及其在肺癌诊断中的价值.方法 用甲基化特异性PCR方法检测组织、血浆及BALF中的PTEN基因启动子区CpG岛甲基化.结果 45例肺癌患者中,PTEN基因启动子异常甲基化率组织为26.67%(12/45)、血浆为15.56%(7/45),BALF为22.22%(10/45);而非肺癌组织、正常对照血浆、非肺癌患者BALF中未检出甲基化;血浆、BALF中甲基化改变与肿瘤组织甲基化状况显著相关(P<0.01).结论 血浆、BALF中PTEN基因异常甲基化改变的检测在肺癌的早期特异诊断等方面有一定的价值.