Clinical–pathologic significance of cancer stem cell marker expression in familial breast cancers

Human breast cancer cells with a CD44⁺/CD24^?/low or ALDH1+ phenotype have been demonstrated to be enriched for cancer stem cells (CSCs) using in vitro and in vivo techniques. The aim of this study was to determine the association between CD44⁺/CD24^?/low and ALDH1 expression with clinical–pathologic tumor characteristics, tumor molecular subtype, and survival in a well characterized collection of familial breast cancer cases. 364 familial breast cancers from the Ontario Familial Breast Cancer Registry (58 BRCA1-associated, 64 BRCA2-associated, and 242 familial non-BRCA1/2 cancers) were studied. Each tumor had a centralized pathology review performed. TMA sections of all tumors were analyzed for the expression of ER, PR, HER2, CK5, CK14, EGFR, CD44, CD24, and ALDH1. The Chi square test or Fisher’s exact test was used to analyze the marker associations with clinical–pathologic tumor variables, molecular subtype and genetic subtype. Analyses of the association of overall survival (OS) with marker status were conducted using Kaplan–Meier plots and log-rank tests. The CD44⁺/CD24^?/low and ALDH1+ phenotypes were identified in 16% and 15% of the familial breast cancer cases, respectively, and associated with high-tumor grade, a high-mitotic count, and component features of the medullary type of breast cancer. CD44⁺/CD24^?/low and ALDH1 expression in this series were further associated with the basal-like molecular subtype and the CD44⁺/CD24^?/low phenotype was independently associated with BRCA1 mutational status. The currently accepted breast CSCs markers are present in a minority of familial breast cancers. Whereas the presence of these markers is correlated with several poor prognostic features and the basal-like subtype of breast cancer, they do not predict OS.     

The expression of β-catenin in different subtypes of breast cancer and its clinical significance

The Wnt/β-catenin signaling pathway is implicated in mammary oncogenesis. Reports of β-catenin expression and its association with outcome in breast cancer are controversial. This study was performed to address the distribution of β-catenin expression in invasive breast cancer and the correlation between β-catenin expression and survival of breast cancer patients, and to determine whether β-catenin was specifically activated in any molecular subtypes. Immunohistochemistry was performed on a tissue microarray containing 169 invasive breast cancers to detect expression of β-catenin. One hundred thirty one of the 169 patients were followed up. Correlation between β-catenin expression and different molecular subtypes was determined using chi-square analysis. Overall survival (OS) was analyzed by Kaplan-Meier method with log-rank test. The invasive breast cancer displayed the different patterns of β-catenin expression from normal tissues with significantly increased cytoplasmic and nuclear staining of β-catenin. Aberrant β-catenin expression was observed in 109 in the 169 cases (64.50 %), and there was no difference in β-catenin expression in the four molecular subtypes. Furthermore, aberrant β-catenin expression was significantly associated with adverse outcome not only in the entire cohort but also in each of the different molecular subtypes. β-catenin activation is preferentially found and is associated with a poor clinical outcome in invasive breast cancer independent of molecular subtype.     

Drug-resistance mechanisms of non-small cell lung cancer to osimertinib and its potent treatment strategies北大核心CSCD

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFRTKIs) have been used in the treatment of patients with non-small cell lung cancer (NSCLC) and epidermal growth factor receptor (EGFR) gene mutations. The first and second-generation of EGFRTKIs have achieved significant results. Osimertinib is one of the third-generation of EGFR-TKIs that has good effects in the patients with the resistance to first-generation of EGFR-TKIs. However, the clinical treatment results show that osimertinib can induce drug resistance. At present, no effective solution has been developed for the drug resistance in clinical application of osimertinib, which has become one of the major obstacles in the treatment of EGFR-positive NSCLC. It has been found that osimertinib treatment can induce the mutations of MET, BIM and other genes, which results in drug resistance of tumor. This drug-resistance can likely be overcome using combination therapies that are targeted to these gene mutation sites. Further studies on the mechanism of resistance to osimertinib will be of great significance in guiding the clinical response to the occurrence of drug resistance. © 2019 by TUMOR All rights reserved.     

Expression of CK5/6 in patients with triple negative breast cancer and its clinical significance北大核心CSCD

Objective: To investigate the expression of cytokeratin 5/6 (CK5/6) in triple-negative breast cancer (TNBC) tissues and its clinical significance. Methods: A total of 347 invasive TNBC patients with complete follow-up accepted operation and confirmed by pathology and immunohistochemistry were collected from January 2010 to December 2012 in Yueyang Hospital of Integrated Traditional Chinese & Western Medicine and Huangpu District Central Hospital. The relationship between CK5/6 expression and clinicopathological features and the effect of CK5/6 on prognosis of TNBC patients were retrospectively analyzed. Results: Of 347 TNBC patients, the expression of CK5/6 was positive in 221 cases (63.8%, 221/347). The expression of CK5/6 was positively correlated with lymph node metastasis (P = 0.027, r = 0.127) and Ki-67 expression (P < 0.001, r = 0.208). The tumor size, clinical stage, lymph node metastasis and CK5/6 were independent prognostic factors of the disease-free survival (DFS) or overall survival (OS) in TNBC patients (all P < 0.05). After the median follow-up of 63 months, the recurrence and metastasis occurred in 21 cases with CK5/6 negative expression, and the death occurred in 15 cases of CK5/6-negative group; while 62 cases of recurrence and metastasis and 52 cases of death were in CK5/6-positive group. The median DFS and OS of TNBC patients in CK5/6-negative group were longer than those in CK5/6-positive group (P = 0.015, P = 0.007). Chemotherapy of anthracycline combined with paclitaxel was beneficial for DFS and OS of TNBC patients in CK5/6-positive group (both P < 0.05), while there was no benefit in CK5/6-negative group (both P > 0.05). Conclusion: The positive expression of CK5/6 is an important risk factor for the prognosis of TNBC patients. The CK5/6-positive patients achieves more benefits in chemotherapy of anthracycline combined with paclitaxel as compared with the CK5/6-negative patients. © 2019 by TUMOR.     

Clinical significance of estrogen receptor β in breast cancer

To estimate the clinical significance of estrogen receptor (ER) β in breast cancer we reviewed some reports and compared them with our preliminary results. The structure of ER β is similar to that of ER α. The DNA binding domain of ER β is 96% conserved compared with ER α, and the ligand binding domain shows 58% conserved residues, suggesting that both receptors can bind estrogen responsive elements on target genes and that they may also bind similar ligand. The target tissues of ER β such as testis, prostate, lung, brain, thymus, and ovary, are different from those of ER α, ovary, uterus, endometrium, and breast. Although the function of ER β in breast cancer progression is not well understood, 30-50% of breast cancers may express ER β mRNA signals. Additionally, ER β may be a useful prognostic factor in patients with breast cancer, because tumors that co-expressed ER α and ER β might be node positive and tend to be of higher grade. Further characterization of the function of ER β and its isoforms in breast cancer is warranted.     

Xanthine oxidoreductase – Clinical significance in colorectal cancer and in vitro expression of the protein in human colon cancer cells

Xanthine oxidoreductase (XOR) is a key enzyme in degradation of DNA and RNA, and has previously been shown to be decreased in aggressive breast and gastric cancer. In this study, XOR expression was assessed in tissue microarray specimens of 478 patients with colorectal cancer and related to clinical parameters. In addition, we performed in vitro studies of XOR activity, protein and mRNA in colon cancer cells (Caco-2). Results from the tissue expression analyses show that XOR was decreased in 62% and undetectable in 22% of the tumours as compared to normal tissue. Loss of XOR was associated with poor grade of differentiation (p = 0.006) and advanced Dukes stage (p = 0.03). In multivariate survival analysis, XOR was a prognostic factor (p = 0.008), independent of Dukes stage, histological grade, age and tumour location. The in vitro analyses show that XOR is not measurable in undifferentiated Caco-2 cells, but appears and increases with differentiation. We conclude that XOR expression is associated with histological grade of differentiation and extent of disease in colorectal cancer, and it provides significant prognostic information independently of established factors.     

Increased expression of enolase α in human breast cancer confers tamoxifen resistance in human breast cancer cells

Enolase-α (ENO-1) is a key glycolytic enzyme that has been used as a diagnostic marker to identify human lung cancers. To investigate the role of ENO-1 in breast cancer diagnosis and therapy, the mRNA levels of ENO-1 in 244 tumor and normal paired tissue samples and 20 laser capture-microdissected cell clusters were examined by quantitative real-time PCR analysis. Increased ENO-1 mRNA expression was preferentially detected in estrogen receptor-positive (ER+) tumors (tumor/normal ratio >90-fold) when compared to ER-negative (tumor/normal ratio >20-fold) tumor tissues. The data presented here demonstrate that those patients whose tumors highly expressed ENO-1 had a poor prognosis with greater tumor size (>2 cm, *P = .017), poor nodal status (N > 3, *P = .018), and a shorter disease-free interval (≦1 year, *P < .009). We also found that higher-expressing ENO-1 tumors confer longer distance relapse (tumor/normal ratio = 82.8–92.4-fold) when compared to locoregional relapse (tumor/normal ratio = 43.4-fold) in postsurgical 4-hydroxy-tamoxifen (4-OHT)-treated ER+ patients (*P = .014). These data imply that changes in tumor ENO-1 levels are related to clinical 4-OHT therapeutic outcome. In vitro studies demonstrated that decreasing ENO-1 expression using small interfering RNA (siRNA) significantly augmented 4-OHT (100 nM)-induced cytotoxicity in tamoxifen-resistant (Tam-R) breast cancer cells. These results suggest that downregulation of ENO-1 could be utilized as a novel pharmacological approach for overcoming 4-OHT resistance in breast cancer therapy.     

β-adrenoceptor signaling and its control of cell replication in MDA-MB-231 human breast cancer cells

MDA-MB-231 human breast cancer cells express high -adrenoceptor levels, predominantly the ₂ subtype. Receptor stimulation by isoproterenol evoked immediate reductions in DNA synthesis which were blocked completely by propranolol and were of the same magnitude as effects elicited by high concentrations of 8-Br-cAMP. Isoproterenol-induced inhibition of DNA synthesis was maintained throughout several days of exposure, resulting in a decrement in total cell number, and the effects were augmented by cotreatment with dexamethason; an even greater effect was seen when cAMP breakdown was inhibited by theophylline, with or without addition of isoproterenol. Despite the persistent effect of isoproterenol, receptor downregulation was evident with as little as 1 h of treatment, and over 90% of the receptors were lost within 24 h. Receptor downregulation was paralleled by homologous desensitization of the adenylyl cyclase response to -adrenoceptor stimulation. Dexamethasone augmented the effects of isoproterenol on DNA synthesis but did not prevent receptor downregulation or desensitization. These results indicate that -adrenoceptors are effectively linked, through cAMP, to the termination of cell replication in MDA-MB-231 human breast cancer cells, and that activation of only a small number of receptors is sufficient for a maximal effect. Novel pharmacologic strategies that focus on cell surface receptors operating through adenylyl cyclase may offer opportunities to combat cancers that are unresponsive to hormonal agents, or that have developed multidrug resistance.     

Clinical characteristics and prognosis of triple-negative breast cancer patients with postoperative initial visceral metastasis北大核心CSCD

Objective: To analyze and discuss the clinicopathologic characteristics of triple-negative breast cancer with initial visceral metastasis after surgery, as well as the efficacy of first-line chemotherapy and the prognostic factors. Methods: A retrospective analysis was performed using the clinical data of 107 triple-negative breast cancer patients with postoperative initial visceral metastasis, who were treated in Department of Medical Oncology, Fudan University Shanghai Cancer Center from January 1, 2011 to June 30, 2013. The analysis also included their first-line chemotherapy efficacy, survival situation and prognostic factors. Results: Among 107 patients, there were 101 patients (94.4%) with invasive ductal carcinoma and 6 patients (5.6%) with other pathological types or mixed patterns. The postoperative median disease-free interval was 14.4 months; the 1-, 2- and 3-year survival rates after visceral metastasis were 75.7%, 41.1% and 22.4%, respectively. The objective response rate of first-line platinum-based chemotherapy was 60.0%, and the median progressionfree survival time was 8.6 months; which were significantly higher or longer than that in the non-platinum chemotherapy group (36.2%, P = 0.014; 5.1 months, P = 0.023). However, there was no significant difference in the median survival time between the two groups (19.9 vs 20.9 months, P = 0.423). Univariate analysis showed that neoadjuvant chemotherapy, postoperative radiotherapy, lymph node metastasis status, clinical stage, disease-free interval, best curative effect of first-line chemotherapy, plurivisceral metastasis, and progression-free survival after first-line chemotherapy had significant effects on the overall survival (all P < 0.05). Additionally, multivariate analysis showed that neoadjuvant chemotherapy and disease-free interval had significant effects on overall survival (both P < 0.05). Conclusion: The triple-negative breast cancer patients with initial visceral metastasis have short disease-free interval time and low long-term survival rate. The first-line platinumbased chemotherapy is a good choice for these patients. The neoadjuvant chemotherapy and disease-free interval within one year may be the independent prognostic factors for their overall survival. Copyright © 2017 by TUMOR All rights reserved.     

Clinical characteristics and prognosis of triple-negative breast cancer patients with postoperative initial visceral metastasis北大核心CSCD

Objective: To analyze and discuss the clinicopathologic characteristics of triple-negative breast cancer with initial visceral metastasis after surgery, as well as the efficacy of first-line chemotherapy and the prognostic factors. Methods: A retrospective analysis was performed using the clinical data of 107 triple-negative breast cancer patients with postoperative initial visceral metastasis, who were treated in Department of Medical Oncology, Fudan University Shanghai Cancer Center from January 1, 2011 to June 30, 2013. The analysis also included their first-line chemotherapy efficacy, survival situation and prognostic factors. Results: Among 107 patients, there were 101 patients (94.4%) with invasive ductal carcinoma and 6 patients (5.6%) with other pathological types or mixed patterns. The postoperative median disease-free interval was 14.4 months; the 1-, 2- and 3-year survival rates after visceral metastasis were 75.7%, 41.1% and 22.4%, respectively. The objective response rate of first-line platinum-based chemotherapy was 60.0%, and the median progressionfree survival time was 8.6 months; which were significantly higher or longer than that in the non-platinum chemotherapy group (36.2%, P = 0.014; 5.1 months, P = 0.023). However, there was no significant difference in the median survival time between the two groups (19.9 vs 20.9 months, P = 0.423). Univariate analysis showed that neoadjuvant chemotherapy, postoperative radiotherapy, lymph node metastasis status, clinical stage, disease-free interval, best curative effect of first-line chemotherapy, plurivisceral metastasis, and progression-free survival after first-line chemotherapy had significant effects on the overall survival (all P < 0.05). Additionally, multivariate analysis showed that neoadjuvant chemotherapy and disease-free interval had significant effects on overall survival (both P < 0.05). Conclusion: The triple-negative breast cancer patients with initial visceral metastasis have short disease-free interval time and low long-term survival rate. The first-line platinumbased chemotherapy is a good choice for these patients. The neoadjuvant chemotherapy and disease-free interval within one year may be the independent prognostic factors for their overall survival. Copyright © 2017 by TUMOR All rights reserved.     

Bone morphogenetic protein-9 regulates expression of long non-coding RNAs in breast cancer cells北大核心CSCD

Objective: To screen out the long non-coding RNAs (LncRNAs) related to bone morphogenetic protein-9 (BMP-9) expression, and to investigate the role of BMP-9 in the growth, differentiation, migration and apoptosis of breast cancer cells by regulating the expression of LncRNA. Methods: MDA-MB-231 cells were infected with the recombinant adenovirus Ad-BMP-9 carrying whole BMP -9 gene (named as MDA-MB-231/BMP-9 cells) or the empty vehicle adenovirus Ad-GFP carrying green fluorescent protein (GFP) gene (named as MDA-MB-231/GFP cells), respectively. Microarray technology was used to detect the difference in LncRNAs expression between MDAMB- 231/BMP-9 and MDA-MB-231/GFP cells. The changes of LncRNA LINC00443, LINC00638, LINC00486, RHNO1, SERHL, HOXA11-AS, IQCA1, LINC00461, LOC440173, LHFPL, ANKRD36BP2, BVES-AS1, LINC00937 and LINC00608 were validated by real-time fluorescent quantitative PCR. The biological funcation and related pathways of the above LncRNAs were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Results: The expression level of BMP-9 mRNA in MDA-MB-231/BMP-9 cells was significantly higher than that in MDA-MB-231/GFP cells (as the control). Compared with the control group, the expression levels of LncRNAs in MDA-MB-231/BMP-9 cells changed significantly. The expression levels of LINC00443, LINC00638 and LINC00486 were up-regulated, while the expression levels of IQCA1, LINC00461, LOC440173, LHFPL and ANKRD36BP2 were down-regulated. The altered LncRNAs participated in the formation of cytoskeleton, cell membrane and so on, or played roles as signal molecules in intercellular signal transduction. Conclusion: The expression profile of LncRNAs in MDA-MB-231 cells with BMP-9 overexpression is significantly changed, indicating that LncRNAs may play key roles in the proliferation and invasion of breast cancer MDA-MB-231 cells regulated by BMP-9. © 2019 by TUMOR.     

Clinical significance of serum GP73, AFP,and CA199 test in the diagnosis of hepatic cancer北大核心CSCD

Objective: To investigate the clinical value of serum Glogi glycoprotein 73 (GP73) and its combination to alpha-fetoprotein (AFP) and CA199 for the diagnosis of primary hepatic cancer (PHC). Methods: Sera from 110 patients (50 hepatic cancer, 20 hepatitis, 20 liver cirrhosis, and 20 malignant tumors of the digestive system except hepatic cancer) and 20 healthy controls were collected. Serum GP73 was analyzed by enzyme-linked immunosorbent assay. The levels of AFP and CA199 were quantified by electrochemiluminescence immunoassays. Results: The median serum GP73 concentration of PHC (104.4 μg/L) was higher than that of hepatitis (22.7 μg/L), liver cirrhosis (53.8 μg/L), malignant tumors of the digestive system except hepatic cancer (41.4 μg/L), and healthy controls (19.3 μg/L) significantly (P<0.05). The sensitivity, specificity, and accuracy to diagnose PHC by GP73 alone were 72.0%, 95.0% and 86.2%, respectively, and were higher than that of AFP or CA199 alone. GP73's sensitivity was highest (92.0%) for the parallel test of GP73, AFP, and CA199; its specificity was highest (100.0%) for the series-wound test of GP73, AFP, and CA199; and its accuracy was the highest (87.7%) for the parallel test of GP73 and AFP. The area under the ROC curve of GP73 was 0.824, which is higher than that of AFP or CA199 alone. The serum GP73 level was not significantly correlated with that of AFP or CA199. Conclusion: GP73 is a new effective marker in the diagnosis of PHC, and the combined examination of GP73, AFP, and CA199 can improve the diagnosis value of PHC.     

Overexpression of β1-chain-containing laminins in capillary basement membranes of human breast cancer and its metastases

Introduction  

Laminins are the major components of vascular and parenchymal basement membranes. We previously documented a switch in the expression of vascular laminins containing the α4 chain from predominantly laminin-9 (α4β2γ1) to predominantly laminin-8 (α4β1γ1) during progression of human brain gliomas to high-grade glioblastoma multiforme. Here, differential expression of laminins was studied in blood vessels and ductal epithelium of the breast.     

TGF-β1 mRNA expression in clinical breast cancer and its relationship to ER mRNA expression

Eighty nine primary breast cancers were investigated forthe expression of TGF-1 and ER mRNA usingPCR of reverse transcribed RNA. PCR products werevalidated using Southern blots and hybridization with radiolabelledcDNA probes. TGF₁ mRNA was found to beexpressed in 56/89 (63%) of the breast cancerswhile ER mRNA was expressed in 23/89 (26%)of the tumours. Using chi-square analysis TGF- mRNAexpression was found to correlate significantly with ERmRNA expression (p < 0.001), in that virtuallyall tumours that expressed ER mRNA co-expressed TGF₁.In tumours that were ER mRNA negative, TGF₁expression was more variable. These results suggest thatduring tumour progression, ER expression is lost morefrequently than is growth factor expression.