乳腺癌变过程中Pim1和Notch1差异性表达的临床价值

目的 探讨Pim1和Notch1在乳腺组织中差异性表达的临床意义及与预后的关系。方法 qRTPCR、免疫组织化学法和数据库检测评估Pim1、Notch1 mRNA和蛋白在乳腺癌旁正常组织、普通型导管增生、导管原位癌和浸润性导管癌中的表达水平, 分析Pim1和Notch1表达与乳腺癌患者临床病理参数的关系,Kaplan-Meier Plotter数据库发掘Pim1和Notch1在乳腺癌预后中的价值。结果 乳腺癌组织中Pim1 mRNA水平升高,Pim1蛋白表达明显降低;Pim1 mRNA与淋巴结转移和TNM晚期密切相关,Pim1蛋白在淋巴结转移及高分级患者中表达显著减少,其低表达的患者无复发生存期更短（P=0.000）。Notch1 mRNA和蛋白在乳腺癌中显著高表达,Notch1 mRNA与淋巴结转移、高分级和晚分期明显相关,Notch1蛋白在淋巴结转移组中表达显著升高,其高表达患者总生存时间减少（P=0.025）。Pim1和Notch1蛋白表达在乳腺癌中负相关（r=-0.385,P=0.001）。结论 Pim1低表达和Notch1高表达可能是促进乳腺癌发生发展及预后不良的危险因素,可能成为评估乳腺癌潜在生物标志物。     

CIP2A expression is elevated in cervical cancer

Early detection of cervical cancer is critical for a favorable prognosis. Standard cytological detection methods, such as Pap smear, are highly subjective and HPV detection is not a reliable marker for predicting the malignancy potential of cervical lesions. As a result, there is a demand for a diagnostic assay capable of sensitive and specific detection of cervical cancer. In this preclinical exploratory study, qRT-PCR and western blotting were used to assess expression levels of CIP2A and p16INK4a in cervical tissue samples (n(normal adjacent) = 23, n(tumor) = 29). CIP2A was abundantly expressed in cervical cancer cell lines and was not expressed in normal epithelial cells. CIP2A mRNA levels were higher in cervical tumor tissues in comparison to the level of CIP2A mRNA in normal adjacent tissue from cervical cancer patients. CIP2A protein was specifically expressed in cervical tumor tissues at different cancer grades and stages, and was not observed in normal adjacent tissue. Elevated CIP2A mRNA levels in cervical tissues had a sensitivity of 80% and specificity of 91% and CIP2A protein expression detection had a sensitivity of 83% and specificity of 100%, similar to that of p16INK4a, with no correlation of CIP2A expression with HPV infection, age, race, or other patient characteristics. However the number of samples analyzed in this preliminary study is limited and a large prospective cohort study is necessary to further evaluate CIP2A as a biomarker for cervical cancer.     

宫颈癌细胞株和宫颈上皮组织中Pin1和Cyclin D1的表达及临床意义

背景与目的：肽基脯氨酰顺反异构酶Pin1在人类多种肿瘤中过表达,Pin1过表达可增强Cyclin D1表达,在肿瘤发生发展中起重要作用。本研究拟探讨宫颈癌细胞株及宫颈上皮组织中Pin1、Cyclin D1的表达及其临床意义。方法：应用RT-PCR、Western blot法检测HeLa、SiHa、C33a、Caski细胞中Pin1和Cyclin D1基因蛋白表达,免疫组化法检测10例正常宫颈、21例宫颈上皮内瘤样病变、57例浸润癌组织中Pin1和Cyclin D1的表达状况,评价其临床意义。结果：HeLa、SiHa、C33a、Caski细胞存在不同程度Pin1 mRNA及蛋白过表达（P〈0．05）。正常宫颈、宫颈上皮内瘤样病变、浸润癌组织中Pin1蛋白表达逐渐增强,分别为0％、47．62％、64．91％（P〈0．05）。Pin1蛋白过表达与宫颈癌临床分期、病理分级、淋巴结转移无关（P〉0．05）;但宫颈腺癌组织中Pin1表达明显高于鳞癌（P〈0．05）。Pin1过表达与Cyclin D1表达呈显著正相关（P〈0．05）。结论：Pin1在宫颈癌细胞株及宫颈癌组织中存在mRNA及蛋白水平上的过表达。Pin1过表达与Cyclin D1表达密切相关．Pin1和Cyclin D1异常表达可能参与宫颈癌的发生发展。     

USP18在宫颈鳞状细胞癌组织中的表达及临床意义

目的:探讨泛素特异性蛋白酶18(ubiquitin specific peptidase 18,USP18)在宫颈鳞状细胞癌组织中的表达及意义,分析USP18的表达水平与宫颈癌临床病例特征的关系.方法:实时荧光定量PCR法检测USP18在30例新鲜宫颈癌组织及癌旁宫颈组织中mRNA的表达情况,蛋白免疫印迹法检测USP1...     

抑癌基因PRDM5在HPV16病毒致宫颈鳞癌发生过程中的作用研究

目的 探讨抑癌基因PRDM5表达下降是否是HPV16病毒感染导致宫颈癌发生的机制之一。方法 采用免疫组化法检测宫颈癌组织及正常宫颈组织中PRDM5蛋白及HPV16 E6/HPV18 E6蛋白的表达;HPV16 E6 shRNA质粒转染SiHa细胞后, 应用RT-PCR及 Western blot法检测PRDM5的表达。结果 HPV16/18 E6在宫颈癌组织中表达阳性率明显高于正常宫颈组织;宫颈癌组织中PRDM5蛋白表达水平较正常宫颈组织中表达水平低;将HPV16 E6 shRNA3转染SiHa细胞干扰HPV16 E6表达后, 发现SiHa细胞中PRDM5 mRNA及蛋白表达上调。结论 PRDM5可能介导HPV16病毒感染导致宫颈癌的发生发展过程。     

Expression profile of histone deacetylases 1, 2 and 3 in ovarian cancer tissues

Objective

To investigate the expression levels of histone deacetylase (HDAC) 1, 2, and 3 in ovarian cancer tissues and normal ovarian tissues.

Methods

Randomly assigned each of six patients with serous, mucinous and endometrioid ovarian cancer were included. Another six patients with normal ovarian tissue were included for comparison. RT-PCR was performed to quantify the levels of HDACs1-3 mRNA in the cancer and normal tissues. Western blot analysis was performed to measure the expression levels of HDACs1-3 protein. The HDACs1-3 expression pattern was also topologically examined by immunohistochemistry.

Results

Increased mRNA expressions of HDCA1, HDAC 2 and HDAC 3 were detected in 83%, 67% and 83% of 18 cancer tissue samples, compared to normal tissue samples. The relative densities of HDAC1 mRNA and HDAC3 mRNA in the serous, mucinous and endometrioid cancer tissues, and HDAC2 mRNA in serous cancer tissues were significantly higher than those of the normal tissues, respectively (p<0.05). Overexpression of HDAC1, HDAC2 and HDAC3 proteins were detected in 94%, 72% and 83% of 18 cancer samples, respectively. The relative densities of HDAC1 protein and HDAC3 protein in serous, mucinous and endometrioid cancer, and HDAC2 protein in serous and mucinous cancer tissues were significantly higher than those of normal tissues, respectively (p<0.05). Most cancer tissues expressed moderate to strong staining of HDACs1, 2 and 3 in immunohistochemistry. Staining of HDAC2 was weak in only one endometrioid cancer tissue.

Conclusion

HDACs1-3 are over expressed in ovarian cancer tissues and probably play a significant role in ovarian carcinogenesis.     

Expressions of IGFBP-5, cFLIP in cervical intraepithelial neoplasia,cervical carcinoma and their clinical significances: a molecular pathology

Background

Insulin-like growth factor binding protein (IGFBPs) have been as potential tumor suppressors in the occurrence and development of tumors. Cellular Fas-associated death domain-like interleukin-1β-converting enzyme (FLICE)-like inhibitory protein (cFLIP) contains a death effect domain (DED), which blocks death receptor pathway and inhibits apoptosis.

Methods

We collected normal cervical tissues from 28 subjects, CIN samples from 37 patients, and cervical cancer tissues from 40 patients. In these samples, we then measured the expression levels of IGFBP-5 and cFLIP via RT-PCR and immunohistochemistry, and we detected the presence of high-risk HPV by Hybrid capture II assays in cervical secretions provided by the subjects.

Results

significant differences in the expression of IGFBP-5 protein among the normal, CIN, and CC tissues (P < 0.05). The highest expression of IGFBP-5 protein was found in CIN stage II and III tissues, whereas the expression of IGFBP-5 in CC samples was decreased relative to controls. The expression level was affected by factors such as clinical stage, pathological differentiation, and lymph node metastasis. Relative to the controls, IGFBP-5 mRNA content was higher in the CC group and lower in the CIN group (P < 0.05). No expression of cFLIP protein or mRNA was detected in normal cervical tissues. However, the degree of pathological changes correlated with increasing expression of cFLIP protein and mRNA, and significant differences were therefore detected between groups (P < 0.05). The HPV infection rates in the CIN and CC groups were much higher than in the normal group (P < 0.05).

Conclusion

IGFBP-5 expression is up-regulated in response to progression of CIN and down-regulated in invasive cervical carcinoma. Detection of IGFBP-5 and cFLIP expression levels, may prove particularly useful for diagnosing and differentiating CIN and CC.     

Promoter methylation of death-associated protein kinase and its role in irradiation response in cervical cancer

This study was aimed at investigating the death-associated protein kinase (DAPK) promoter methylation and its clinical relevance in cervical cancer. The DAPK promoter methylation was detected by methylation-specific PCR (MSP) and correlated with DAPK mRNA and protein expression. The effect of DAPK expression on the radiosensitivity of the cervical cancer cell line was assessed by overexpressing DAPK in the radioresistant cell line SiHa. DAPK hypermethylation was found in 56.08% of the cervical cancer samples and was associated with the tumor histological cell type of squamous cell carcinoma (p=0.002) and advanced tumor stage (p=0.005). Subsequently, DAPK protein expression was found to significantly decrease in cervical cancer samples when compared to normal tissues. The DAPK mRNA and protein expression levels were absent or remarkably reduced in SiHa and HeLa in which the DAPK promoter was hypermethylated. The expression levels of DAPK could be restored after demethylation treatment with 5-aza-2'-deoxycytidine. Overexpressing DAPK in vitro had no significant influence to the survival of the radioresistant SiHa cell after being challenged by irradiation. Our findings suggest that DAPK might not directly be responsible for the cellular radiosensitivity, however, DAPK hypermethylation appeared to be of prognostic significance in the advanced stages of cervical cancer.     

DYRK1B蛋白在宫颈癌组织中的表达及意义

目的 探讨双特异性酪氨酸磷酸化调节激酶1B(DYRK1B)蛋白在宫颈癌组织中的表达情况及其临床意义.方法 选取84例宫颈癌病理组织标本(宫颈癌组)、40例宫颈上皮内瘤变(CIN)组织标本(CIN组)、40例正常宫颈组织标本(对照组),采用免疫组织化学染色方法检测3组标本中DYRK1B蛋白的表达情况,并探讨DYRK1B蛋白表达与宫颈癌病理特征的关系.结果 宫颈癌组中DYRK1B蛋白的阳性表达率为72.62%,高于CIN组中的22.50%和对照组的12.50%(P﹤0.05);CIN组和对照组中的DYRK1B蛋白的阳性表达率比较,差异无统计学意义(P﹥0.05);不同临床分期、组织浸润深度、分化程度、淋巴结转移情况的宫颈癌组织中DYRK1B蛋白的阳性表达率比较,差异均有统计学意义(P﹤0.05);而不同年龄、病理学类型的宫颈癌组织中DYRK1B蛋白的阳性表达率比较,差异均无统计学意义(P﹥0.05).结论 DYRK1B蛋白在宫颈癌组织中表达上调,可能与宫颈癌的发生发展有关.     

叉头框蛋白Q1在宫颈鳞癌中的表达及其临床意义

目的:探讨叉头框蛋白Q1(forkhead box Q1,FOXQ1)在宫颈鳞癌细胞系及组织中的表达及其与患者临床病理特征和预后的关系。方法:采用RT-PCR和Western blot检测宫颈鳞癌细胞系SiHa和CaSki及人宫颈永生化鳞状细胞系Ect1/E6E7中FOXQ1 mRNA和蛋白的表达情况;分别应用RT-PCR及免疫组化法检测60例宫颈鳞癌组织中FOXQ1 mRNA和蛋白的表达情况;进一步采用Log-rank检验和Cox回归模型对宫颈鳞癌患者的预后因素进行分析,并用Kaplan-Meier计算不同FOXQ1表达情况下患者的生存率曲线。结果:与Ect1/E6E7相比,SiHa和CaSki中FOXQ1 mRNA和蛋白的表达水平均显著上调（P<0.05）;宫颈鳞癌组织中FOXQ1 mRNA的表达水平显著高于配对癌旁组织（3.096±0.109 vs 0.902±0.040,P<0.01）;免疫组化结果显示,FOXQ1蛋白在宫颈鳞癌组织中高表达,阳性率为68.33%,且FOXQ1蛋白的表达与肿瘤浸润深度、淋巴结转移、淋巴脉管浸润及FIGO分期有关（P<0.05）,而与患者的年龄、肿瘤分化程度和肿瘤大小无关（P<0.05）;Kaplan-Meier 单因素分析显示,FOXQ1蛋白是宫颈鳞癌患者的重要预后因素（P<0.05）,进一步Cox多因素回归分析显示FOXQ1蛋白是影响宫颈鳞癌患者的独立预后因素（HR=2.703,95%CI:1.081~6.758,P<0.05）。结论:FOXQ1的高表达可能与宫颈鳞癌的发生、发展及预后关系密切,有望成为评估宫颈鳞癌预后的有效指标。     

极光激酶A在宫颈鳞状细胞癌患者血清中的表达及其临床意义

目的:应用生物信息学技术分析和筛选影响宫颈癌(CC)预后的关键基因并探讨极光激酶A(AURKA)在宫颈鳞状细胞癌(CSCC)中的表达及其临床意义。方法:从基因表达综合(GEO)数据库下载并分析CC患者的mRNA表达谱及临床数据,利用生物信息学技术筛选出CC组织与正常宫颈组织间的差异表达基因(DEGs)。PPI网络和生存分析结果显示AURKA基因的表达和CC分期及患者预后相关。利用GEPIA2、HPA数据库分别验证AURKA mRNA及蛋白在正常宫颈组织和不同病理分期CC组织中的表达情况。采用Kaplan-Meier法分析CC患者的AURKA基因高、低表达(以中位数为截断值)与生存期是否相关。随后进一步通过实时荧光定量PCR(qPCR)和酶联免疫吸附试验(ELISA)技术分别检测在CSCC、宫颈上皮内瘤变(CIN)和女性健康体检者血清中AURKA mRNA及蛋白的表达。结果:GEPIA2数据库分析结果显示AURKA mRNA在CC组织中高表达(P<0.05),且AURKA mRNA在CC不同阶段的表达水平存在显著差异(P<0.05),随着肿瘤分期的进展AURKA mRNA的表达升高。HPA数据库分析结果显示,与正常宫颈组织相比,AURKA蛋白表达水平在CC组织中亦升高,差异具有统计学意义(P<0.05)。Kaplan-Meier生存分析结果显示,AURKA基因高表达的CC患者生存时间更短。qPCR和ELISA实验显示,CSCC和CIN患者血清AURKA mRNA的相对表达水平明显高于健康体检者(P<0.05)。CSCC患者血清AURKA蛋白的表达水平明显高于CIN及健康体检者(P<0.05)。AURKA mRNA及蛋白在CC血清中均高表达。ROC曲线及χ^(2)检验分析结果显示AURKA mRNA的诊断价值较高,血清AURKA mRNA高表达组患者FIGO分期、淋巴结转移、肿瘤体积及肌层浸润深度均分别高于AURKA mRNA低表达组(P<0.05)。结论:CSCC患者血清中AURKA mRNA及蛋白的表达水平升高,两者联合检测对CSCC的诊断及预后监测具有潜在应用价值。     

JAG1和Notch3在乳腺癌组织中的表达及预后价值

目的探讨JAG1和Notch3在乳腺癌组织中的表达水平及预后意义。方法通过在线Oncomine数据库和Kaplan-Meier Plotter数据库分析JAG1和Notch3在乳腺癌组织中的表达情况及对预后的影响。收集2008年1月至2013年12月乳腺浸润性导管癌组织及配对癌旁组织各80例,采用实时荧光定量PCR(QPCR)和免疫组织化学SP法分别检测JAG1和Notch3的mRNA和蛋白水平,分析JAG1和Notch3蛋白表达与乳腺癌临床病理特征和总生存时间(OS)的关系。多因素分析采用Cox比例风险回归模型。结果 Oncomine数据库分析显示,乳腺癌组织中JAG1和Notch3的mRNA水平均高于癌旁组织(P<0.05)。80例乳腺浸润性导管癌组织中JAG1 mRNA水平为1.58±0.56,高于癌旁组织的1.23±0.48(P=0.0039);乳腺浸润性导管癌组织中Notch3 mRNA水平为2.39±0.83,高于癌旁组织的1.95±0.64(P=0.0036)。JAG1和Notch3蛋白在乳腺癌组织中的高表达率为71.3%(57/80)和80.0%(64/80),分别高于癌旁组织的8.8%(7/80)和11.2%(9/80),差异均有统计学意义(P=0.005,P=0.003). JAG1和Notch3蛋白表达在乳腺癌组织中呈正相关(r=0.267,P=0.017)。JAG1和Notch3表达与乳腺癌淋巴结转移、TNM分期和组织学分级有关(P<0.05)。Kaplan-Meier Plotter在线分析显示, JAG1或Notch3高表达的乳腺癌患者的OS短于低表达者(P>0.05)。80例乳腺浸润性导管癌患者中,JAG1高表达者的中位OS为38.3个月,低于JAG1低表达者的80.4个月(P=0.0394);Notch3高表达者的中位OS为39.3个月,低于Notch3低表达者的80.2个月(P=0.0334),且JAG1和Notch3均高表达者的中位OS最短,为27.3个月。Cox多因素分析显示,淋巴结转移、JAG1表达和Notch3表达是影响乳腺癌患者OS的独立因素(P<0.05)。结论 JAG1和Notch3在乳腺癌组织中高表达,且与预后不良有关,有望成为临床评估乳腺癌预后的潜在生物标志物。     

3-磷酸甘油酸脱氢酶在宫颈鳞状细胞癌中高表达

目的:检测宫颈鳞状细胞癌组织和细胞中3-磷酸甘油酸脱氢酶(3-phosphoglycerate dehydrogen-ase,PHGDH)的表达水平,探讨其在肿瘤发生发展中的作用.方法:使用实时荧光定量PCR法和免疫组织化学法检测宫颈鳞状细胞癌和正常宫颈组织中PHGDH mRNA转录水平和蛋白翻译水平;利用实时荧光定量...     

Endo-sulfatase Sulf-1 protein expression is down-regulated in gastric cancer

In our recent report on gene expression in gastric cancer we identified the endo-sulfatase Sulf-1 gene to be up-regulated in gastric tumors relative to apparently normal (AN), and paired normal (PN) gastric tissue samples. In the present report we investigate the protein expression levels of Sulf-1 gene in gastric tumors, AN and PN samples using tissue microarray (TMA) and immunohistochemistry. Expression data was collected from two sets of TMA's containing replicate sections of tissue samples. Scoring data from TMA set-1 revealed a significant difference in Sulf-1 immunoreactivity between tumors and "normals" (PN and AN) (p-value = 0.001928). Also, Sulf-1 expression in tumors was also significantly different from either PN (p-value = 0.019) or AN (p-value = 0.006) samples. Similar results were obtained from analysis of scoring data from the second set of arrays. Comparison of mRNA expression and protein expression in gastric tumor tissues revealed that in 6/20 (30%) tumor samples showed up-regulated protein expression concordant with over-expression of mRNA. However, a discord with mRNA being over-expressed relative to down regulated protein expression was observed in majority 14/20 (70%) of tumor samples. Our study indicates down regulation of Sulf-1 protein expression in gastric tumors relative to PN and AN samples which is discordant with mRNA over-expression seen in tumors.