High survivin mRNA expression is a predictor of poor prognosis in breast cancer: a comparative study at the mRNA and protein level

Background

Survivin plays a key role in the initiation and progression of breast cancer. However, its prognostic relevance to breast cancer patients has long been a matter of debate. The purpose of this study was to examine the expression of survivin and its role in predicting clinical outcome in a series of human breast cancer cases both at the mRNA and protein level.

Methods

Formalin-fixed paraffin-embedded tumor tissues from 245 female patients with invasive breast cancer and 13 patients with ductal carcinoma in situ were examined for survivin mRNA by quantitative real-time RT-PCR (RT-qPCR). In addition, 237 of these tumors with invasive breast cancer were available for immunohistochemistry (IHC). The relationship between survivin status and clinicopathological characteristics and prognosis was evaluated.

Results

RT-qPCR revealed that high levels of survivin mRNA were strongly associated with high nuclear grade, positive axillary lymph nodes, negative hormone receptor status, positive Her2 amplification, higher Ki67 labeling index, and presence of vascular invasion. In the Cox proportional regression model analysis, survivin mRNA was shown to be a significant univariate parameter for relapse-free survival (RFS), distant relapse-free survival (DRFS), and breast cancer-specific survival (BCSS) as well as a significant multivariate parameter for RFS, DRFS, and BCSS. In hormone receptor (HR)-positive/Her2-negative subtype cases, survivin mRNA expression was also an independent predictor in terms of DRFS. Immunohistochemically, positive staining was seen in the cytoplasm and/or nucleus of cancer cells, although this did not correlate with the mRNA level, and harbored no prognostic value.

Conclusions

High mRNA expression of survivin was an independent marker of poor prognosis both in the entire cohort and in the HR-positive/Her2-negative subtype, whereas the protein expression of survivin was not. These findings suggest that RT-qPCR can provide more reliable data than IHC in validating the prognostic significance of survivin for breast cancer patients.     

Cullin1 is a novel marker of poor prognosis and a potential therapeutic target in human breast cancer

BackgroundTo investigate the role of Cullin1 (Cul1) in the development of breast cancer, we examined the expression of Cul1 in breast cancer tissues and analyzed the correlation between Cul1 expression and clinicopathologic variables and patients survival.Patients and methodsWe evaluated the Cul1 expression by immunohistochemistry using a tissue microarray (TMA) which includes 393 breast cancer tissues. We also studied the role of Cul1 in breast cancer cell proliferation, migration and invasion by carrying out CCK8 cell proliferation assay, cell migration and invasion assay.ResultsThe Cul1 expression was significantly correlated with breast cancer histology grade (P = 0.000), estrogen receptor status (P = 0.001), progesterone receptor status (P = 0.001) and human epidermal growth factor receptor 2 status (P = 0.002). Furthermore, we showed a strong correlation between high Cul1 expression and worse 5-year overall and disease-specific survival rates in breast cancer patients (P = 0.026 and P = 0.015, respectively). Finally, we found that Cul1 knockdown inhibits cell proliferation, migration and invasion abilities.ConclusionsCul1 overexpression is significantly correlated with breast cancer progression and predicts worse survival. Cul1 regulates breast cancer cell proliferation, migration and invasion.     

High BRCA2 mRNA expression predicts poor prognosis in breast cancer patients

The prognostic significance of BRCA2 mRNA levels in tumor tissues was studied in sporadic breast cancer patients. BRCA2 mRNA levels were determined by real-time PCR. Histologic grade III tumors showed significantly (p = 0.001) higher BRCA2 mRNA levels (0.828 +/- 0.102 BRCA2/beta-glucuronidase mRNA ratio, mean +/- SE) than histologic grade I and II tumors (0.438 +/- 0.055) and estrogen receptor (ER)-negative tumors (0.773 +/- 0.102) showed a nonsignificant (p = 0.072) trend toward an increase in BRCA2 mRNA levels compared to ER-positive tumors (0.541 +/- 0.079). Other clinicopathologic parameters, such as menopausal status, lymph node status and tumor size, were not significantly associated with BRCA2 mRNA levels. Patients with high BRCA2 mRNA levels showed a significantly (p = 0.006) lower 5-year disease free survival rate (63%) than those with low levels (94%). Lymph node metastases, ER negativity and high histologic grade were also significantly (p < 0.05) associated with poor prognosis. Multivariate analysis revealed that BRCA2 mRNA levels were a significant prognostic factor, being independent of the other conventional prognostic factors. Our results suggest that BRCA2 mRNA levels might serve as a clinically useful prognostic factor in breast cancer patients.     

An evaluation of immunoreactivity for c-erbB-2 protein as a marker of poor short-term prognosis in breast cancer

Eighty-five breast carcinomas from the same number of patients have been assessed immunohistochemically using the antiserum 21N for the presence of the c-erbB-2 protein. Twenty-two of the patients had evidence of advanced disease (tumour fixation or distant metastases) at presentation. Follow-up was for a median of 24 months. c-erbB-2 protein was detected in the majority of cells in 14 (16.5%) carcinomas, and to a lesser extent in a further six (7%) tumours. There was no relationship between staining and stage, node status or size but more poorly differentiated carcinomas had evidence of staining (36%) than well (17%) or moderately (14%) differentiated carcinomas (P = 0.02). There was a significant association between staining and mortality (P = 0.009) and recurrence (P = 0.0002). The relative risk of death for staining compared to no staining (after adjusting for node status, stage and grade) was 2.97 (95% confidence interval 1.29, 6.84) and the relative risk of recurrence for staining compared to no staining after similar adjustment was 3.85 (95% confidence interval 1.86-7.97). In this particular group of patients immunoreactivity for c-erbB-2 protein is an independent indicator of poor short-term prognosis.     

Expression of neuronal protein synuclein gamma gene as a novel marker for breast cancer prognosis

Synucleins are emerging as central players in the fundamental neural processes and in the formation of pathologically insoluble deposits characteristic of neurodegenerative diseases. However, synuclein γ (SNCG), previously identified as a breast cancer specific gene (BCSG1), is also highly expressed in breast carcinomas, but not expressed in normal or benign breast tissues. We analyzed SNCG gene expression in 93 clinical breast specimens and associated it with clinical outcome. Overall SNCG mRNA expression was detectable in 36% breast cancers. However, 81% of stage III/IV breast cancers were positive for SNCG expression, while only 15% of stage I/II breast cancers were positive for SNCG expression. In contrast, SNCG was undetectable in benign breast lesions. Expression of SNCG in the primary tumor also significantly associated with lymph node involvement and metastasis. There was no significant correlation between SNCG gene expression and age, menstruation, and status of ER, PR, PCNA, and HER-2. Patients whose tumors expressed SNCG had a significantly shorter DFS and a high probability of death when compared with those whose tumors did not express SNCG. The hazard ratio of metastasis or recurrence according to the SNCG status was 4.515 (95% CI, 1,188–17.154; P = 0.027). Cox multivariate analysis showed that SNCG had independent prognostic significance above and beyond conventional variables. This study suggests that the expression of SNCG is an independent predictive marker for recurrence and metastasis in breast cancer progression. SNCG is expected to be a useful marker for breast cancer progression and a potential target for breast cancer treatment.     

The pS2 gene, mRNA, and protein: a potential marker for human breast cancer

Approximately 50% of human breast tumors secrete a small cysteine-rich protein called pS2. In the human breast cancer cell line MCF-7, expression of the pS2 protein is strongly induced by estrogen, and cloning and sequence analysis of the pS2 gene has revealed an "estrogen responsive element" in the gene's 5'-flanking region. The results of immunohistochemical assays and radioimmunoassays on breast cancer biopsies indicate that the pS2 protein is a marker for hormone-dependent breast tumors and that its expression is associated with longer overall, and disease-free, survival. The pS2 protein is also expressed in normal stomach mucosa and in regenerative tissues in ulcerative diseases of the gastrointestinal tract. Its physiological function is unknown.     

Expression of Y-Box-binding protein 1 in Chinese patients with breast cancer

The purpose of this study was to investigate the expression of Y-Box-binding protein 1 (YB-1) in breast cancer and its correlation with clinicopathological characteristics and prognosis. Paraffin sections were retrospectively collected from 239 cases of stage I–III breast cancer patients and 30 healthy females who received surgery between January 2000 and December 2004 in the Chinese People’s Liberation Army General Hospital. The protein expression of YB-1 was detected by immunohistochemistry. The expression difference between the two groups and the correlation between YB-1 expression and clinicopathological characteristics and breast cancer prognosis were analyzed. Within the breast cancer group, YB-1 was expressed in the cytoplasm in 100.0% (239/239) of cases and in the nucleus in 36.8% (88/239) of cases. Within the control group of normal breast tissue, YB-1 was expressed in the cytoplasm in 100.0% (30/30) of cases and in the nucleus in 16.7% (5/30) of cases. The expression of YB-1 in the nucleus of breast cancer cells was significantly higher than that in normal breast tissue (P = 0.029). The expression of YB-1 in the nucleus of breast cancer cells positively correlated with the Scarff–Bloom–Richardson grade (P = 0.007) and HER-2 expression (P = 0.005), negatively correlated with ER expression (P = 0.004), and was independent of the age, menstrual status, pathological type, tumor size, lymph node status, presence of thrombosis, PR expression, and EGFR expression. The 5-year disease-free survival (DFS) and overall survival (OS) of patients with positive YB-1 expression in the nucleus were significantly lower than those of patients who were negative for nuclear YB-1 expression, and the difference was statistically significant (DFS 65.9% vs. 82.1%, P = 0.000; OS 79.5% vs. 92.1%, P = 0.000). Multivariate analysis suggested that the expression of YB-1 in the nucleus is an independent prognostic factor that affects DFS and OS in breast cancer patients (DFS P = 0.015; OS P = 0.035). In conclusion, the expression of YB-1 in the nucleus is related to carcinogenesis and the development of breast cancer. Therefore, YB-1 is an important molecular marker that can be used to predict breast cancer prognosis.     

Low RAI2 expression is a marker of poor prognosis in breast cancer

Breast Cancer Research and Treatment - Retinoic acid-induced 2 (RAI2) has been shown to be a putative suppressor of the early hematogenous dissemination of tumor cells to the bone marrow in breast...     

Thymidine kinase 1 immunoassay: a potential marker for breast cancer

Previous research indicates that thymidine kinase I (TKI) possesses value as a tool for both prognosis and diagnosis in breast cancer. However, drawbacks to the existing radioassay for thymidine kinase have frustrated its clinical use. To overcome these drawbacks, we developed a monoclonal antibody to TK1. We have assessed this antibody for a linear antibody-antigen response and for reproducibility using ELISA techniques. We also have evaluated this antibody for TKI specificity as determined by Western blot. To test the accuracy of this monoclonal antibody further, we treated human MCF-7 breast cancer cells with tamoxifen and measured decreasing TKI activity and protein levels with the radioassay and with our monoclonal antibody in an ELISA, respectively. We then used the radioassay and our monoclonal antibody to measure TK1 activity and protein levels, respectively, in 218 serum samples of postoperative breast cancer patients and found a correlation between the two assays. Our results demonstrated that the TK1 immunoassay not only had a linear, reproducible, and specific response but accurately measured TK1 levels in both MCF-7 breast cancer cells and serum. Thus, our monoclonal antibody may demonstrate potential for practical use in a clinical setting for the management of breast cancer.     

Expression of CLDN1 in colorectal cancer: a novel marker for prognosis

Claudin1 (CLDN1) plays an important role not only in the intercellular barrier function of tight junctions (TJs) but also in migration and invasiveness of cancer cells. Previous reports show that CLDN1 overexpression is significantly related to the malignant behavior in several cancer types whereas its significance in colorectal cancer (CRC) is not fully understood. The present study comprised 119 patients who underwent surgery for CRC, as well as 3 cell lines derived from human CRC. The correlation of gene expression with clinical parameters in patients was assessed by knockdown experiments using 3 cell lines. Patients with high CLDN1 expression were statistically shown to have a relatively better prognosis, and those with low CLDN1 expression showed poorer overall survival and disease-free survival than those with high expression. The assessment of CLDN1 knockdown in the 3 cell lines demonstrated that the siRNA inhibition resulted in a statistically significant increase in cell invasiveness. In conclusion, the present data strongly suggest that CLDN1 expression is a prognostic factor in CRC patients.     

MAGE-D4B is a novel marker of poor prognosis and potential therapeutic target involved in breast cancer tumorigenesis

Melanoma-associated antigen (MAGE) family members are generally described as tumor-specific antigens. An association between MAGE-D4B and breast cancer has yet to be reported and the functional role of the encoded protein has never been established. We performed microarray analysis of 104 invasive breast tumors and matched non-cancerous breast biopsies. qPCR was used for validation in an independent biobank. To investigate the biological relevance of MAGE-D4B in breast tumorigenesis, its phenotypic effects were assessed in vitro. Overall, MAGE-D4B was detected in 43% of tumors while undetected in normal breast tissue. MAGE-D4B was found to correlate with tumor progression and to be an independent prognostic marker for poor outcome in term of relapse-free and overall survival, with potential predictive relevance in relation to response to chemotherapy. RNA interference-mediated knockdown of MAGE-D4B significantly hampered the invasive properties of Hs578T cells by affecting anchorage-independent growth, adhesion, migration and invasion affecting anchorage-independent growth, adhesion, migration and invasion and by modulating expression of invasion-suppressor gene E-cadherin.     

Overexpression of cyclinD1 predicts for poor prognosis in estrogen receptor-negative breast cancer patients

CyclinD1 plays a critical role in regulating cell cycle progression. CyclinD1 mRNA and protein are overexpressed in approximately 50% of primary breast cancer cases. However, its clinical significance as a predictive factor remains unclear. One hundred and seventy-three female patients diagnosed with invasive ductal carcinoma who had undergone a mastectomy (161 patients) or breast-conserving surgery (12 patients) were followed up for 6-119 months (median 86 months) postoperatively. Immunoreactivity for monoclonal anti-cyclinD1 antibody (clone DCS-6) with paraffin-embedded carcinoma tissues was investigated using a labeled streptavidin-biotin method. Overexpression of cyclinD1 was found in 42% (73 of 173), and strongly correlated with estrogen receptor (ER) expression (p < 0.000001). Univariate analysis revealed no association between overexpression of cyclinD1 and overall survival or relapse-free survival in all patient groups. However, in the ER-negative subgroup (n = 75), overexpression of cyclinD1 was significantly correlated with shorter overall survival (p = 0.018) and relapse-free survival (p = 0.014) as well as the lymph node status and tumor size. In contrast, there were no significant associations between overexpression of cyclinD1 and clinical outcome in the ER-positive subgroup. According to Cox's multivariate analysis in the ER-negative subgroup, overexpression of cyclinD1 had the most significant effect on overall survival (p = 0.02) and relapse-free survival (p = 0.0058), followed by nodal status and histologic grade. These findings suggest that overexpression of cyclinD1 is an independent prognostic indicator in ER-negative breast cancer patients.     

High PD-L1 expression was associated with poor prognosis in 870 Chinese patients with breast cancer

Background

To investigate the role of PD-L1 expression in tumor recurrence and metastasis in Chinese patients with breast cancer.

Methods

Suitable tissue samples were available from 870 patients with breast cancer. Paraffin-embedded tumor sections were stained with PD-L1 antibody. The correlations between PD-L1 expression and clinical characteristics, ER/PR/HER2 status and survival parameters were analyzed. Kaplan-Meier and univariate Cox proportional hazards model analyses were used to compare the survival of patients with high PD-L1 expression and patients with no PD-L1 expression.

Results

The median follow-up time was 98 months(range, 17–265 months). The positive rate of PD-L1 expression in breast cancer was 21.7% (189/870). PD-L1 high expression was inversely associated with larger tumor size, higher tumor grade, more positive lymph node number, as well as negative ER and PR status. PD-L1 expression was particularly higher in TNBC compared with non-TNBC, although no statistical significance was observed. Nomogram logistic regression results based on clinical and pathological features showed that the following factors were more likely associated with high PD-L1 expression: patient age younger than 35 years, larger tumor size, lymphovascular invasion and advanced stage. Our data indicated that patients with high PD-L1 expression had poor DFS, DMFS and overall survival compared with those with no PD-L1 expression. Univariate Cox proportional hazards model analysis showed that PD-L1 was an independent prognostic factor for tumor prognosis.

Conclusions

PD-L1 expression is an important indicator of unfavorable prognosis in breast cancer patients.     

Hypoploidy defines patients with poor prognosis in breast cancer

The clinicopathological features currently used in breast cancer prognosis often fail to characterize the clinical heterogeneity of the disease accurately. Our study is aimed to investigate the predictive value of DNA flow cytometry in breast cancer. Previously untreated breast carcinoma samples (584) were snap frozen for flow-cytometry. Tumors were classified into three DNA index (DI) categories: i) tumors showing a DI =0.96-1.15 (diploid and near-diploid); ii) tumors with a DI >or=1.16 (hyperdiploid, tetraploid, multiploid and/or those with more than one diploid population); and iii) tumors with a DI 相似文献   

High Wilms' tumor 1 mRNA expression correlates with basal-like and ERBB2 molecular subtypes and poor prognosis of breast cancer

The role of Wilms' tumor 1 (WT1) in breast cancer and the relationship between WT1 expression and clinicopathological factors, molecular subtypes and prognosis of breast cancer patients have not been clarified to date. We used publicly available microarray datasets of 266 early breast cancer patients to perform bioinformatics analysis on the relationship between WT1 mRNA expression and breast cancer. Results showed that WT1 mRNA expression was correlated with higher histological grades, ER-negative and basal-like and ERBB2 molecular subtypes in breast cancer. With regard to disease-free survival analysis, the WT1 high expression group showed worse prognosis than the low expression group in univariate analysis, and WT1 was demonstrated to be an independent prognostic indicator in multivariate analysis. This study confirms an oncogenic role of WT1 and demonstrates a possible relation between WT1 and progression of breast cancer.     

FKBPL: a marker of good prognosis in breast cancer

FK506-binding protein-like (FKBPL) has established roles as an anti-tumor protein, with a therapeutic peptide based on this protein, ALM201, shortly entering phase I/II clinical trials. Here, we evaluated FKBPL''s prognostic ability in primary breast cancer tissue, represented on tissue microarrays (TMA) from 3277 women recruited into five independent retrospective studies, using immunohistochemistry (IHC). In a meta-analysis, FKBPL levels were a significant predictor of BCSS; low FKBPL levels indicated poorer breast cancer specific survival (BCSS) (hazard ratio (HR) = 1.30, 95% confidence interval (CI) 1.14–1.49, p < 0.001). The prognostic impact of FKBPL remained significant after adjusting for other known prognostic factors (HR = 1.25, 95% CI 1.07–1.45, p = 0.004). For the sub-groups of 2365 estrogen receptor (ER) positive patients and 1649 tamoxifen treated patients, FKBPL was significantly associated with BCSS (HR = 1.34, 95% CI 1.13–1.58, p < 0.001, and HR = 1.25, 95% CI 1.04–1.49, p = 0.02, respectively). A univariate analysis revealed that FKBPL was also a significant predictor of relapse free interval (RFI) within the ER positive patient group, but it was only borderline significant within the smaller tamoxifen treated patient group (HR = 1.32 95% CI 1.05–1.65, p = 0.02 and HR = 1.23 95% CI 0.99–1.54, p = 0.06, respectively). The data suggests a role for FKBPL as a prognostic factor for BCSS, with the potential to be routinely evaluated within the clinic.     

散发性乳腺癌中DJ-1蛋白高表达与不良预后

目的 探讨DJ-1蛋白表达水平在散发性乳腺癌发生、发展及预后中的临床意义.方法 采用免疫组织化学法检测了包含199例散发性乳腺癌组织与160例乳腺良性增生组织的组织微阵列芯片中DJ-1蛋白的表达情况,同时应用酶联免疫吸附试验检测48例散发性乳腺癌及35例乳腺良性增生患者血清中DJ-1蛋白的水平,并分析组织中DJ-1蛋白表达水平与乳腺癌患者临床病理指标及生存资料的相关性.统计分析采用Pearson ×^2检验和独立样本t检验;生存分析采用log-rank检验绘制Kaplan-Meier曲线,Cox回归用于进行多变量分析.结果 DJ-1蛋白在乳腺良性增生组织及乳腺癌组织中阳性表达率分别为33.8％ （54/160）、48.7％（97/199）,在乳腺良性增生及乳腺癌患者血清中DJ-1蛋白平均水平分别为（25.86±7.28） ng/ml和（31.31±8.39） ng/ml,与乳腺良性增生相比,乳腺癌组织（×^2=8.182,P=0.004）及血清（t=3.161,P=0.002）中DJ-1蛋白的表达水平均显著增加;淋巴结转移阳性的乳腺癌组织与阴性组织相比,DJ-1表达水平显著增加（×^2=5.372,P=0.020）;随着组织学分级的增加,浸润性导管癌中DJ-1表达水平显著增高（×^2=5.244,P=0.022）;DJ-1阳性表达与患者生存期缩短显著相关（OS：×^2=5.427,P=0.020;×^2=4.606,DFS：P=0.032）.单变量Cox回归分析发现,DJ-1表达水平作为危险因素与患者的OS及DFS显著相关（OS：P=0.023; DFS：P=0.036）,但多变量Cox回归分析发现,DJ-1表达水平并非影响患者预后的独立风险因素（OS：P=0.561; DFS：P=0.342）.结论 在散发性乳腺癌中,DJ-1蛋白表达水平显著增高,其高表达与乳腺癌的侵袭转移、病程进展及不良预后相关.