Prognostic Indicator In Breast Cancer Research Articles

Relationships between mutant P53 protein overexpression and other prognostic indicators in breast cancer

Relationships between mutant P53 protein overexpression and other prognostic indicators in breast cancer

Tumor Suppressor p53 Mutations and Breast Cancer: A Critical Analysis

Alterations in the tumor suppressor gene p53 are the most commonly identified changes in cancer, including neoplasia of the breast. The activity of p53 is regulated post-translationally. Phosphorylation state, subcellular localization, and interaction with any of a number of cellular proteins are likely to influence the function of p53. The exact effect of p53-mediated growth suppression seems to be cell-type specific but appears to be directly related to the ability of p53 to act as a specific transcriptional activator. The role that transcriptional repression plays in the function of WT p53 is less clear. It is also possible that p53 has a more direct activity in DNA replication and repair. Most documented p53 mutations result in single amino acid substitutions which may confer one or more of a spectrum of transforming abilities on the protein. Mutation may lead to nuclear accumulation of p53 protein; however, inactivation of p53 by nuclear exclusion and interaction with the mdm2 protein also appear to be important in tumorigenesis. Used in conjunction with other established factors, accumulation of cellular p53 may be a useful prognostic indicator in breast cancer. A syngeneic mouse model system yielded evidence that p53 mutations are important in the early, preneoplastic stages of mammary tumorigenesis. This murine system may provide the ability to investigate the functions of p53 in the early stages of breast cancer which are technically difficult to examine in the human system.

Immunoreactive prostate‐specific antigen in lung tumors

Prostate-specific antigen (PSA) is a glycoprotein produced by the epithelial cells of the prostate. PSA is currently used clinically to diagnose and monitor prostate carcinoma. In previous work we have demonstrated that 30% of breast tumors and, more rarely other tumors, contain significant amounts of PSA. PSA appears to be a favorable prognostic indicator in breast cancer. Here, using a sensitive assay, we demonstrated for the first time that lung adenocarcinomas and squamous cell carcinomas also contain PSA. PSA in lung tumor extracts was present mainly in its 33 KDa form (free PSA), at levels measurable by commercial methods. The presence of PSA was associated more closely with male patients and adenocarcinomas. The physiological role of PSA in lung tissue and the prognostic significance of PSA in lung cancer remain to be determined. These and our previous data as well as reports by other groups support the view that PSA is a ubiquitous biochemical marker of steroid hormone action.

Association of vascular endothelial growth factor expression with tumor angiogenesis and with early relapse in primary breast cancer.

Angiogenesis is an independent prognostic indicator in breast cancer. In this report, the relationship between expression of vascular endothclial growth factor (VEGF; a selective mitogen for endothelial cells) and the microvessel density was examined in 103 primary breast cancers. The expression of VEGF was evaluated by immunocytochemical staining using anti‐VEGF antibody. The microvessel density, which was determined by immunostaining for factor VIII antigen, in VEGF‐rich tumors was clearly higher than that in VEGF‐poor tumors (P<0.01). There was a good correlation between VEGF expression and the increment of microvessel density. Furthermore, postoperative survey demonstrated that the relapse‐free survival rate of VEGF‐rich tumors was significantly worse than that of VEGF‐poor tumors. It was suggested that the expression of VEGF is closely associated with the promotion of angiogenesis and with early relapse in primary breast cancer.

Quantitative analysis of mutant p53 protein in breast tumor cytosols and study of its association with other biochemical prognostic indicators in breast cancer.

Breast tumors are thought to originate, grow, and metastasize in an environment which includes steroid hormone receptors, their cognate steroid ligands, and many gene products which are regulated by steroid hormone receptor-ligand complexes. In this paper we describe highly sensitive and quantitative immunofluorometric procedures for measuring three proteins that are candidate prognostic indicators in breast cancer, namely, the p53 tumor suppressor gene product, carcinoembryonic antigen (CEA), and prostate specific antigen (PSA). These proteins were quantified in over 950 cytosolic tumor extracts along with estrogen and progesterone receptors (ER, PR). Association analysis between all five biochemical parameters revealed strong negative associations between p53 and receptors and strong positive associations between CEA and receptors. Negative associations between p53 and CEA and between CEA and PSA were also found. These associations, not quantitatively studied in previous reports, are related to each other using a hypothetical model. The observed associations may further contribute to the understanding of the biology of breast tumors.

Flow cytometry: potential utility in monitoring drug effects in breast cancer.

Flow cytometric analysis of DNA ploidy and S-phase fraction are well recognized prognostic indicators in breast cancer. The present paper deals with the widening of the applications of flow cytometry to monitoring the effectiveness of antiestrogen therapy, detecting clonal selection and emergence of drug resistance, and monitoring chemosensitizing properties of drugs. Antiestrogen activity can be studied by DNA flow cytometry to address clinical research problems such as patient-specific pharmacokinetics, dosing compliance, and acquired antiestrogen resistance. Patient plasma specimens containing various concentrations of triphenylethylenes can be monitored for drug-induced effects using cell cycle measurements and correlated to in vivo drug levels. DNA flow cytometry has also been instrumental in the study of the effects of prolonged low-dose (0.5 microM for > 100 days) tamoxifen treatment on human estrogen receptor negative MDA-MB-231 cells, where it was shown that tamoxifen may significantly alter cell cycle kinetics and tumorigenicity of these cells, selecting a new, more aggressive, and rapidly growing clone. Lastly, it has been shown that the chemosensitizing properties of another triphenylethylene antiestrogen, toremifene, on estrogen receptor negative, multidrug resistant MDA-MB-231-A1 human breast cancer cells can be studied using flow cytometric analysis. Toremifene (and its metabolites N-desmethyltoremifene and toremifene IV) are able to "resensitize" MDA-MB-231-A1 cells to vinblastine and doxorubicin, as reflected in a marked shift of cells to G2/M phase of the cell cycle. Flow cytometry is a widely available technique that might be applied clinically to monitor, at the cellular level, drug effects on tumors, including the modulators of drug resistance.

Ectopic production of prostate specific antigen by a breast tumor metastatic to the ovary.

We have recently reported that about 30% of breast tumors produce prostate specific antigen (PSA). We examined here, 99 primary ovarian cancer tumors and found relatively low levels of PSA in only three tumors. One patient with metastatic ovarian cancer from a primary breast tumor, produced relatively high levels of PSA and responded well to antiestrogen treatment although she was steroid receptor-negative. Another patient with metastatic ovarian cancer from a primary breast tumor did not produce PSA and did not respond to treatment although she was steroid hormone receptor-positive. This data describe for the first time, ectopic PSA production by a breast tumor at the ovarian metastatic site and further support the view that PSA is a favourable prognostic indicator in breast cancer.

Prediction of early relapse and shortened survival in patients with breast cancer by proliferating cell nuclear antigen score

Cell cycle kinetic measures have been shown to have prognostic significance in breast cancer. Methods that have been used to assess the proliferating fraction of tumors include measurements of DNA content with S-phase calculation by flow cytometric analysis, radioisotope-labeled nucleotide incorporation, and cell cycle-associated protein expression. The recent discovery of the S-phase-specific nuclear protein proliferating cell nuclear antigen (PCNA) opens the door for a novel approach to cell kinetic measurement with an immunocytochemical assay. A quantitative immunocytochemical assay for PCNA was performed on 82 primary invasive breast carcinomas fixed in formaldehyde solution and embedded in paraffin and 18 corresponding axillary metastases. The percentage of tumor cells with strong nuclear staining was determined by visual count. This PCNA score was correlated with histologic parameters, age, relapse intervals, and long-term survival. PCNA scores were distributed normally among primary carcinomas (range, 5-54; mean, 22.5). Carcinomas had much higher scores than adjacent normal epithelium (means, 22.5 and 4.1, respectively; P < 0.00001), and axillary node metastases had slightly higher scores than corresponding primary breast tumors (means 26.4 and 22.5, respectively; P = 0.05). The PCNA score did not correlate with age, tumor size, axillary node status, intramammary lymphatic-vascular invasion, or estrogen and progesterone binding capacities. Furthermore, its variability could not be explained by grade. PCNA values increased as the mitotic rate increased. Cancers with high PCNA scores (> or = 25) were associated with shorter disease-free (P = 0.007) and overall survival times (P = 0.01) than tumors with low PCNA scores (< 25) (median follow-up, 166 months). PCNA score has potential value as a prognostic indicator in breast cancer. This method of assessing the proliferating pool offers advantages over other assays in terms of relative simplicity of the method, applicability to paraffin-embedded fixed tissue, and low cost.

Neu(c-erbB-2/HER2) and the epidermal growth factor receptor (EGFR) in breast cancer.

One hundred and eighty thousand new cases of invasive breast cancer were diagnosed in 1992 within the United States. This disease affects approximately 1 out of 8 women in the US. Chemotherapy and/or hormonal therapy have shown some improved disease-free and/or overall survival rates. Unfortunately, this type of therapy is not directed specifically to the malignant cells, and systemic toxicities are observed. In order to develop site-specific treatment, the biology of the disease must be understood such that certain genes or their products which are involved in the pathogenesis of the disease can be targeted. Two structurally related tyrosine kinase growth factors, the epidermal growth factor receptor (EGFR) and c-erbB-2 (neu) have been identified in human breast cancer tissue and, in many instances, may function as oncogenes. The clinical data related to these two growth factor receptors as prognostic factors for the disease have been critically evaluated. Several problems with the critical studies were identified, and solutions were proposed to clarify the conflicting results reported in the studies which have attempted to examine whether c-erbB-2 (neu), in particular, is a prognostic indicator for breast cancer. In addition, data related to the structure of, ligands for and interaction between the proteins have been reviewed and presented with respect to their role in breast cancer development. A more thorough understanding of the genetic changes which contribute to the development of breast cancer will lead to more specific and less toxic treatment for this disease.

Genetic markers as prognostic indicators in breast cancer.

Identifying markers that have the potential to predict tumor behavior is important in breast cancer because of the variability in clinical disease progression. Genetic alterations in tumors may appear as changes in total DNA content, individual chromosomes, single genes, or gene expression. Alteration in DNA content is an imprecise but accessible measurement of the genome. Diploid tumors have been associated with a better clinical outcome, and increased ploidy correlates with other indicators of poor prognosis. Concurrent analysis of DNA content with markers of genetic expression is feasible (e.g., myc oncogene) and may increase its prognostic power. Chromosomal studies could provide a more precise tool for localizing genetic damage, but there is little cytogenetic information about primary breast cancers, no convincing evidence has emerged to target locations in the karyotype that appear specifically altered, and many primary and cultured breast cancers contain cells that appear chromosomally normal. Attempts to define molecular markers have used probes of different chromosomal sites, some chosen because of logical associations with hormonal activity, known oncogenes, or tumor-suppressor genes, and some by chance. Currently, to the authors' knowledge, none has shown uniform changes by mutation, loss, or overexpression in all breast cancers, although a remarkable number of loci are altered to some extent. These lesions must be associated with particular disease subsets or, retrospectively, with differential survival if they are to have prognostic value. The authors examined several loci (ERBB2, INT2, MUC1) for gene amplification or loss of heterozygosity by Southern blotting and for gene expression by immunohistochemistry in breast tumors from patient groups selected by survival. A retrospective series showed gene amplification at the erbB2 locus in 22% of rapidly recurrent (RR) tumors and 13% of tumors from long-term tumor-free survivors (LTS), but the difference was not statistically significant (P = 0.18). The erbB2 product was displayed histochemically with equal frequency between those with RR tumors and LTS patients. Moreover, the correlation was poor between different analytic measures on the same tumors. This result was tested using a prospective study of erbB2 to correlate DNA analysis with western blot findings and frozen and fixed histochemical results. Another oncogene, int2, showed significant correlation between amplification and recurrence; 16% of RR tumors showing genetic amplification (P = 0.02). Loci on other chromosomes, 1 (muc1) and 17 (cmm86), also are being investigated in groups selected for differences in survival.

Long-term prognostic significance of thymidine labelling index in primary breast cancer.

Tumour growth rates, as measured by incorporation of tritiated thymidine, have been reported as being of prognostic importance in breast cancer. We have measured the thymidine labelling index (TLI) of 185 early breast cancers, followed-up for a minimum of 8 years. Above median TLI was associated with higher tumour grade, but not with other prognostic factors. TLI was not predictive of survival in either univariate or multivariate analysis. The inter- and intra-observer reproducibilities of TLI measurements were poor, which may be a factor limiting its usefulness as a prognostic indicator in breast cancer.

DNA ploidy and breast cancer-A retrospective study of paraffin-embedded tissues wtth 5 year follow-up

Over the past 7 years increasing demand has occurred for better prognostic Indicators of breast cancer. Besides the traditional factors of tumour size, type, grade and lymph node status, oestrogen receptor assay has now become a mandatory laboratory assay and there is increasing clinical demand for DNA quantitation, cell cycle analyses with S-phase fraction studies, MDR markers and proliferative activity. At Cauifield General Medical Centre we have studied 86 consecutive breast biopsies in which a diagnosis of carcinoma was made in 1985/*86. In each case frozen section reports matched the final paraffin section assessment. In 79 cases a diagnosis of in-situ or infiltrating duct carcinoma was made. Two cases each of infiltrating lobular, mucinous and papillary carcinoma and a single medullary carcinoma was identified. In each case a single block of paraffin-embedded formalin - fixed tissue was selected tor DNA pioidy assessment. The tissue was processed by a method slightly modified from Hedleys original description in 1983. The single cell suspension was stained with propidlum Iodide and analysed In a Coulter Epics Profile II. The smallest cell yield was 2750 cells and the largest 45,000. The mean CV was 7.1. DNA ploldy and S-phase fraction assessments were performed using Phoenix Multicyle software on an Arrow 386, 33 mHz computer with an Intel 387 Chip. Of the 86 carcinomas 46 were DNA diploid, 23 aneuploid and 17 tetraploid. Age distribution showed peak incidence in 7th & 8th decades with a mean tumour size of 22mm. Over half the patients were node negative (49/86). In our series survival curves show patients with Infiltrating duct carcinomas who present in the 6th & 7th decade who are node negative and have diploid tumours have a far superior 5 year survival (70%). Over the past 7 years increasing demand has occurred for better prognostic Indicators of breast cancer. Besides the traditional factors of tumour size, type, grade and lymph node status, oestrogen receptor assay has now become a mandatory laboratory assay and there is increasing clinical demand for DNA quantitation, cell cycle analyses with S-phase fraction studies, MDR markers and proliferative activity. At Cauifield General Medical Centre we have studied 86 consecutive breast biopsies in which a diagnosis of carcinoma was made in 1985/*86. In each case frozen section reports matched the final paraffin section assessment. In 79 cases a diagnosis of in-situ or infiltrating duct carcinoma was made. Two cases each of infiltrating lobular, mucinous and papillary carcinoma and a single medullary carcinoma was identified. In each case a single block of paraffin-embedded formalin - fixed tissue was selected tor DNA pioidy assessment. The tissue was processed by a method slightly modified from Hedleys original description in 1983. The single cell suspension was stained with propidlum Iodide and analysed In a Coulter Epics Profile II. The smallest cell yield was 2750 cells and the largest 45,000. The mean CV was 7.1. DNA ploldy and S-phase fraction assessments were performed using Phoenix Multicyle software on an Arrow 386, 33 mHz computer with an Intel 387 Chip. Of the 86 carcinomas 46 were DNA diploid, 23 aneuploid and 17 tetraploid. Age distribution showed peak incidence in 7th & 8th decades with a mean tumour size of 22mm. Over half the patients were node negative (49/86). In our series survival curves show patients with Infiltrating duct carcinomas who present in the 6th & 7th decade who are node negative and have diploid tumours have a far superior 5 year survival (70%).

乳癌サーモグラフィ所見と予後の検討;サーモグラフィにおける温度上昇と血管構築の変化

乳癌サーモグラフィ所見と予後の検討;サーモグラフィにおける温度上昇と血管構築の変化

Epidermal growth factor receptor expression as a prognostic indicator in breast cancer

The significance of epidermal growth factor receptor (EGFR) status as a prognostic indicator was investigated by a competitive binding assay in 135 primary breast cancer patients. 55 patients (41%) were EGFR positive and EGFR status was negatively correlated with oestrogen receptor (ER) status ( P < 0.01). 5-year postoperative follow-up showed that relapse-free survival for EGFR positive patients was significantly worse than that for EGFR negative patients ( P < 0.05). There was no difference between the two groups in tumour size, axillary node involvement, age and menopausal status. Analysis by axillary node status demonstrated the poor prognosis of the EGFR positive group in node positive patients. As yet, no difference in prognosis has been seen in node negative patients. A higher frequency of haematopoietic relapse was observed in EGFR positive patients. Simultaneous or sequential EGFR measurements in primary tumour and metastatic sites of 34 patients showed that expression of EGFR was more enhanced in metastatic sites.

Is aryl hydrocarbon hydroxylase activity a new prognostic indicator for breast cancer?

Aryl hydrocarbon hydroxylase (AHH) activity was measured in the breast tumours of 153 primary and 17 recurrent cancer patients, and in 18 patients with benign breast tumour. All operations were carried out in 1983-84. The cytosolic fraction was collected for steroid receptor determination, and microsomes were separated for AHH assay from the same tissue samples. The AHH distribution was wide and highly skewed in all groups. About 10% of the samples showed activities below detection limit. The medians and ranges for primary cancers were 34 (less than 5-2683), for recurrent cancers 40 (20-239) and for benign tumours 11 (less than 5-37) fmol min-1 mg-1 protein. After logarithmic transformation, the mean AHH activities of cancer samples differed significantly from those of benign tumours. The logarithm of AHH activity (log AHH) correlates positively with axillary lymph node status, and negatively with steroid receptor levels. The development of the disease and the survival of the patients were followed for 4 years. The survival and the disease-free interval of the cancer patients who had low AHH activity was significantly higher than that of the high AHH group. The multivariate analysis with Cox's proportional hazad model showed primary tumour size, progesterone receptor concentration, nodal status and log AHH to be the most important independent prognostic factors for survival, while the occurrence of metastases, log AHH and tumour size were the equivalent factors for the disease-free interval in primary breast cancers. We conclude that AHH activity may reflect the overall malignant potential of breast cancer tissue.

Prognostic factors in resectable breast cancer.

The traditional unfavorable clinicopathologic features in operable breast cancer are large tumor size, number of axillary node metastases, poorly differentiated grade, presence of lymphatic tumor emboli near to the primary malignancy, blood vessel invasion, tumor necrosis, intense lymphoplasmocytic reaction around the tumor and perimenopausal status. Current multidisciplinary approach is still based on two major clinico-pathologic findings such as tumor size and number of infiltrated axillary nodes. Only in recent years the presence or absence of estrogen receptors was taken into consideration to administer adjuvant tamoxifen or adjuvant chemotherapy, respectively. Since the fundamental prognostic indicators in breast cancer are the total tumor cell number and its inherent biological aggressiveness, it is important to assess the clinical role of the proposed new determinants as a guide to prognosis in series of consecutive patients staged and managed according to uniform treatment programs. Properties such as hormone receptor status, ploidy and tumor cell kinetics (3H-TdR labeling index and percent of S-phase cell) as well as oncogenes should be evaluated also as expression of tumor cell burden and/or indicators of clinical aggressiveness. Recent results from retrospective case series strongly suggest that tumor cell proliferative activity, in particular labelling index, is a prognostic factor independent of axillary nodes, tumor size, and receptor status for the relapse-free survival of node-positive and node-negative tumors. This new prognostic factor should be taken into consideration in the selection of candidates for adjuvant chemotherapy.

A potential radioiodinated ligand for androgen receptor: 7.alpha.-methyl-17.alpha.-[2'-(E)-iodovinyl]-19-nortestosterone

The presence of androgen receptor (AR) in prostate cancer has been linked to the androgen-dependent nature of the tumor and has also been shown to have prognostic significance; it also appears to be a positive prognostic indicator in breast cancer. However, due to the relatively low AR concentrations in most tumors and the inherently low specific activity of tritium, the assay of AR based on available 3H-ligands is not sensitive enough to measure accurately the amount of receptor in small specimens. A 125I-ligand like those available for the estrogen and progesterone receptors would be helpful, but development of such a ligand for AR has not been very successful. Although several androgen analogues containing iodine, bromine, or selenium have been synthesized specifically as potential probes for AR, none have shown any significant affinity or specificity for the receptor. We therefore undertook the synthesis of new potential AR ligands which could be radioiodinated, and determined their affinities for AR (from rat uterus and MCF-7 human breast cancer cells) by using a competition assay. We have examined both 5 alpha-dihydrotestosterone (5 alpha-DHT) and 19-nortestosterone analogues and have identified two such compounds which showed high AR affinity: (17 alpha,20E)-17 beta-hydroxy-21-iodo-5 alpha-pregn-20-en-3-one (17 alpha-[E)-iodovinyl)-5 alpha-DHT, 9) and 17 beta-hydroxy-7 alpha-methyl-(17 alpha,20E)-21-iodo-19-norpregna-4,20-dien-3- one (7 alpha-methyl-17 alpha-[E)-iodovinyl)-19-nortestosterone, 11). In fact, the affinity of the latter for human AR was found to be superior to that of 5 alpha-DHT itself. These iodovinyl analogues could be easily prepared in the radioiodinated form, and should prove to be extremely useful in assaying low levels of AR in small specimens.

The biology of metastatic breast cancer

Early detection and surgical removal of breast cancer are most effective in managing a disease that may affect up to one in ten women in North America and Western Europe. However, one of the most important prognostic indicators for breast cancer is the presence of neoplastic cells in the axillary lymph nodes. The dissemination of cells from a primary lesion, resulting in the progressive growth of metastatic carcinoma in distant sites (including bone, lungs, liver, and brain) is the most common cause of death in breast cancer patients. Experimental studies on the biology of metastatic breast cancer have used rodent tumor systems, and, in recent years, the transplantation of human breast carcinoma cells into athymic mice. The results of such studies, combined with clinical observations, suggest that metastasis is not a random event. The formation of secondary lesions is the result of a sequence of selective events. A better understanding of the metastatic phenotype from cellular and molecular analyses will provide a basis for rational approaches to preventing and treating this most lethal aspect of breast cancer.

Inverse Relationship Between Estrogen Receptor and Epidermal Growth Factor Receptor mRNA Levels in Human Breast Cancer Cell Lines

Epidermal growth factor receptors (EGF-R) are present in a number of human breast cancer cell lines and tumor biopsies. Furthermore, it has been suggested that EGF-R levels are higher in estrogen receptor negative (ER-) than in ER+ human breast tumors and that EGF-R status may be a prognostic indicator in breast cancer. The present study was undertaken to establish whether there is a quantitative relationship between EGF-R and ER mRNA concentrations in a series of 10 well-characterized human breast cancer cell lines. All cell lines expressed detectable quantities of EGF-R mRNA by Northern analysis but the relative abundance of EGF-R mRNA varied more than 50-fold. Two transcripts corresponding to the 10.5- and 5.8-kb mRNAs described in other cell types were present but in different relative proportions in different cell lines. When these lines were divided into an ER+ and an ER- group based on their ability to bind estradiol, ER- cell lines were shown to express significantly higher concentrations of EGF-R mRNA than did ER+ cell lines (p less than 0.005). Furthermore, linear-regression analysis revealed a significant inverse relationship between ER and EGF-R mRNA concentrations both within the group of 10 human breast cancer cell lines as a whole (r = 0.66) and within the 6 functionally ER + lines (r = 0.77). This demonstration of a significant (p less than 0.005) inverse relationship between the concentrations of ER and EGF-R mRNAs in ER + cell lines raises the possibility of reciprocal regulation of the expression of these genes in human breast cancer.

Relation of tumor size, lymph node status, and survival in 24,740 breast cancer cases.

Two of the most important prognostic indicators for breast cancer are tumor size and extent of axillary lymph node involvement. Data on 24,740 cases recorded in the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute were used to evaluate the breast cancer survival experience in a representative sample of women from the United States. Actuarial (life table) methods were used to investigate the 5-year relative survival rates in cases with known operative/pathologic axillary lymph node status and primary tumor diameter. Survival rates varied from 45.5% for tumor diameters equal to or greater than 5 cm with positive axillary nodes to 96.3% for tumors less than 2 cm and with no involved nodes. The relation between tumor size and lymph node status was investigated in detail. Tumor diameter and lymph node status were found to act as independent but additive prognostic indicators. As tumor size increased, survival decreased regardless of lymph node status; and as lymph node involvement increased, survival status also decreased regardless of tumor size. A linear relation was found between tumor diameter and the percent of cases with positive lymph node involvement. The results of our analyses suggest that disease progression to distant sites does not occur exclusively via the axillary lymph nodes, but rather that lymph node status serves as an indicator of the tumor's ability to spread.