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What we know 2000-To read more from these articles from 2000 Click Here.

2000: Primary inflammatory carcinoma of the breast: retrospective review of mammographic findings.

AJR Am J Roentgenol 2000 Feb;174(2):535-8, Kushwaha AC, et al.

“OBJECTIVE: Our goal was to describe the mammographic characteristics of primary inflammatory carcinoma of

the breast. MATERIALS AND METHODS: We identified the medical records of 43 women who participated in a

chemotherapy protocol for primary inflammatory carcinoma of the breast between 1994 and 1997. Mammograms

were available for review in 26 women (age range, 34-78 years; mean age, 56 years). Two radiologists

independently reviewed the 26 mammograms obtained before patients underwent treatment. A third observer was

the final arbiter when needed. RESULTS: Mammographic findings included skin thickening in 24 patients (92%),

diffusely increased density in 21 patients (81%), trabecular thickening in 16 patients (62%), axillary

lymphadenopathy in 15 patients (58%), architectural distortion or focal asymmetric density in 13 patients

(50%), and nipple retraction in 10 patients (38%). Malignant-appearing calcifications were seen in six

patients (23%), and a mass was seen in four patients (15%). CONCLUSION: Diffuse mammographic abnormalities

such as skin thickening, increased density, trabecular thickening, and axillary lymphadenopathy are common

at presentation in patients with primary inflammatory carcinoma of the breast. Mammographic masses and

malignant-appearing calcifications are uncommon manifestations of this disease.

2000: The Role of the RhoC GTPase Oncogene in Inflammatory Breast Cancer.

AACR Poster 4533, van Golen, KL, et al.

“Inflammatory breast cancer (IBC) is an extremely aggressive and metastatic form of breast cancer that has a

5-year disease-free survival rate of less than 45%. Little is known about the genetic components of the

disease. We identified 17 genes by differential display comparing the SUM149 IBC cell line with two normal

human mammary epithelial (HME) cell lines, and the patients own lymphocytes. Twenty-nine IBC and 19 stage-

matched non-inflammatory breast tumors from patients were probed for expression of these genes by in situ

hybridization. Two genes were found to be concordantly altered in 91% of IBC versus 0% of the non-

inflammatory samples analyzed. RhoC GTPase, a putative oncogene, was found to be over-expressed, while

expression of a novel gene, LIBC, was found to be lost in most of the IBC samples analyzed. To understand

what role these genes play in the metastatic IBC phenotype, stable transfectants of HME cells over-

expressing RhoC were established. The transfectants were found to form 10-100-fold more colonies in soft

agar than control-transfected HME cells. And, in a Matrigel invasion assay had, on average, a 5-fold

increase in invasion over the control cells. When evaluated for production of angiogenic factors, the RhoC

transfectants produced VEGF to ~90% the level of the SUM149 IBC cell line. Preliminary in vivo data

indicates that the HME RhoC transfectants are able to form tumors in nude mice. Taken together these data

support that RhoC acts as an oncogene in IBC and likely is a key determinant of it s metastatic phenotype.”

2000: Esthetic reconstruction after mastectomy for inflammatory breast cancer: is it worthwhile?”

J Am Coll Surg 2000 Mar;190(3):304-9, Chin PL, et al.

“BACKGROUND: Because inflammatory breast cancer (IBC) has been viewed as a malignancy with a poor likelihood

of longterm survival, few women have been offered esthetic reconstruction after mastectomy for IBC. Recent

advances in multimodality therapy have improved the outcomes for women with this disease. The purpose of

this review was to assess the results of esthetic breast reconstruction in the population with IBC. STUDY

DESIGN: Review of medical records at the City of Hope National Medical Center for the 10-year period ending

in May 1997, revealed 23 women who underwent elective esthetic breast reconstruction after mastectomy for

IBC. The records of these patients were reviewed retrospectively. Patients requiring reconstruction for

large surgical chest wall defects were not included in the review. RESULTS: Treatment for IBC included

mastectomy in all patients, chemotherapy in 22, and chest wall radiation therapy in 14. Immediate

reconstruction was performed at the time of mastectomy (n = 14) or was delayed (n = 9). The types of

reconstruction included transverse rectus abdominis musculocutaneous flap (n = 18), latissimus dorsi flap (n

= 2), or prosthetic mammary implant reconstruction (n = 3). Seven women chose to undergo additional

reconstruction procedures (ie, nipple reconstruction) after their initial reconstruction. With a median

followup of 44 months for survivors, 16 patients developed recurrence after reconstruction. Of these, 6 were

local recurrences and 10 were distant failures. Seven patients are currently alive with no evidence of

disease, 4 are currently alive with disease, and 12 have died as a result of breast cancer. The median

disease-free survival after reconstruction was 19 months. The median overall survival after reconstruction

for all patients was 22 months. The only negative predictor of survival was a positive surgical margin at

mastectomy. CONCLUSIONS: The significant emotional and esthetic benefits of breast reconstruction should be

available to women with IBC. In light of the improving prognosis of IBC with current aggressive

multimodality treatment, reconstructive procedures should be offered as part of comprehensive therapy.”

2000: RhoC GTPase, a novel transforming oncogene for human mammary epithelial cells that partially

recapitulates the inflammatory breast cancer phenotype.

Cancer Res 2000 Oct 15;60(20):5832-8, van Golen, KL, et al.

“Inflammatory breast cancer (IBC) is the most aggressive form of breast cancer and is phenotypically

distinct from other forms of locally advanced breast cancer. In a previous study, we identified specific

genetic alterations of IBC that could account for a highly invasive phenotype. RhoC GTPase was overexpressed

in 90% of IBC archival tumor samples, but not in stage-matched, non-IBC tumors. To study the role of RhoC

GTPase in contributing to an IBC-like phenotype, we generated stable transfectants of human mammary

epithelial cells overexpressing the RhoC gene. The HME-RhoC transfectants formed large colonies under

anchorage-independent growth conditions, were more motile, and were invasive. In conjunction with an

increase in motility, overexpression of RhoC led to an increase in actin stress fiber and focal adhesion

contact formation. Furthermore, orthotopic injection into immunocompromised mice led to tumor formation.

Taken together, these data indicate that RhoC GTPase is a transforming oncogene in human mammary epithelial

cells and can lead to a highly invasive phenotype, akin to that seen in IBC.”

2000: Inflammatory breast cancer survival: the role of obesity and menopausal status at diagnosis.

Breast Cancer Res Treat 2000 Nov;64(2):157-63, Chang, S, et al.

“No previous studies have evaluated the effect of body size and menopausal status at diagnosis on survival

from inflammatory breast cancer (IBC). We evaluated whether obesity and menopausal status had an impact on

IBC survival in a cohort of 177 female IBC patients seen from 1974 to 1993 at The University of Texas MD

Anderson Cancer Center. Survival time was defined as time from diagnosis until death or censorship at last

date of contact. We categorized women by body size by using the National Institutes of Health/National

Heart, Lung, and Blood Institute’s definitions of obesity as body mass index ((BMI) = weight in kg/(height

in m)2) > or = 30, overweight as 25 < or = BMI < 30kg/m2, and normal/lean as BMI < 25 kg/m2. Cox

proportional hazards analysis, adjusting for axillary lymph node involvement and chemotherapy protocol,

revealed a modifying effect of menopausal status at diagnosis on the association between obesity and IBC

survival (P = 0.02). Relative to postmenopausal women, premenopausal women had significantly worse survival

(hazard ratio (HR) = 1.51, 95% confidence interval (CI) = 1.03-2.22). After stratifying by menopausal

status, premenopausal obese women had non-significantly better survival than their leaner premenopausal

counterparts (HR = 0.63, 95% CI = 0.34-1.15) while postmenopausal obese women had significantly worse

survival than their leaner counterparts (HR = 1.86, 95% CI = 1.02-3.40). These findings suggest that factors

associated with larger body size at diagnosis may contribute to shorter IBC survival among postmenopausal

women but not premenopausal women, who were found to have poorer survival regardless of body size.”

2000: Inflammatory breast carcinoma: pathological or clinical entity?

Breast Cancer Res Treat 2000 Dec;64(3):269-73, Amparo RS, et al.

“Inflammatory breast carcinoma (IBC) diagnosis is usually based in the presence of typical clinical symptoms

(redness and edema in more than 2/3 of the breast), which are not always associated with pathologic

characteristics (subdermal lymphatics involvement). Whether exclusively pathologic findings without clinical

symptoms are sufficient for IBC diagnosis remains controversial. A retrospective analysis of 163 clinically

diagnosed IBC (CIC) either with dermal lymphatics invasion or not, was compared with another group of 99

patients with dermal lymphatics invasion without clinical symptoms (occult inflammatory carcinoma) (OIC).

The following clinical and pathological characteristics have been analyzed and compared: age, menopausal

status, clinical axillar node involvement, symptoms duration before diagnosis, grade, estrogen receptors,

presence of metastases at diagnosis, local recurrence, metastasic dissemination, disease-free (DFS) and

overall survival (OS). Median age was younger in CIC (52.3 vs. 63.8 years; p < 0.001). Symptom duration

before diagnosis were significantly shorter in CIC (3.4 vs. 6.8 months: p < 0.0001). Visceral (36.2% vs.

17.2%; p = 0.001) and brain metastases (7.4% vs. 1%; p = 0.02) was significantly more frequent in CIC.

Negative estrogen receptors were more frequent in CIC (34.9% vs. 65.1%: p < 0.004). Five-years DFS (25.6 vs.

51.6%; p < 0.0001) and OS (28.6 vs. 40%; p < 0.05) were shorter in CIC. CIC (regardless of subdermal

lymphatics involvement) must be clearly differentiated from OIC. Prognosis of CIC patients is poorer, so

this two entities should be clearly differentiated when therepeutic results are reported.”

2000: Molecular biology of breast cancer metastasis. Inflammatory breast cancer: clinical syndrome and

molecular determinants.

Breast Cancer Res 2000;2(6):423-9, Kleer, CG, et al.

“Inflammatory breast cancer (IBC) is an aggressive form of locally advanced breast cancer (LABC) that

effects approximately 5% of women with breast cancer annually in the USA. It is a clinically and

pathologically distinct form of LABC that is particularly fast growing, invasive, and angiogenic. Nearly all

women have lymph node involvement at the time of diagnosis, and approximately 36% have gross distant

metastases. Despite recent advances in multimodality treatments, the prognosis of patients with IBC is poor,

with a median disease-free survival of less than 2.5 years. Recent work on the genetic determinants that

underlie the IBC phenotype has led to the identification of genes that are involved in the development and

progression of this disease. This work has been aided by the establishment of primary human cell lines and

animal models. These advances suggest novel targets for future interventions in the diagnosis and treatment

of IBC.”

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2001: Absence of endothelial cells, central necrosis, and fibrosis are associated with aggressive

inflammatory breast cancer.

Cancer Res 2001 Jan 15;61(2):445-51, Shirakawa K, et al.

“We recently established a new human inflammatory breast cancer (IBC) xenograft (WIBC-9) originating from a

patient with IBC. The graft was transplantable in BALB/c nude and severe combined immunodeficient (SCID)

mice. WIBC-9 was frequently accompanied by lung metastasis and exhibited erythema of the overlying skin,

reflecting its human counterpart. Histological study of the original tumor and WIBC-9 revealed invasive

ductal carcinoma with a hypervascular structure of solid nests and marked lymphatic permeation in the

overlying dermis. In the central part of the solid nests, absence of endothelial cells, central necrosis,

and fibrosis were observed. In vitro, WIBC-9 formed tube-like structures and loops, reflecting its in vivo

feature and its human counterpart. WIBC-9 exhibited aneuploidy, ErbB-2 gene amplification, and an absence of

estrogen receptor and progesterone receptor, which is consistent with IBC. Comparative studies of WIBC-9,

three established non-IBC xenografts, and a human breast cancer cell line (SK-BR3) by reverse transcription

-PCR, ELISA, and immunohistochemistry indicated that certain human genes (interleukin 8, vascular epidermal

growth factor, basic fibroblast growth factor, angiopoietin 13, Flt-1, Tie-2, and Tie-1) and certain murine

genes (integrin alpha(v)beta3, flt-1, tie-2, vascular epidermal growth factor, and CD31) were overexpressed

in exposure to tumor cells. The molecular basis and these unique histological features may be associated

with aggressive IBC on angiogenic and nonangiogenic pathways.”

2001: Persistent E-cadherin expression in inflammatory breast cancer.

Mod Pathol 2001 May;14(5):458-64, Kleer CG, et al.

“E-cadherin is a transmembrane glycoprotein that mediates epithelial cell-to-cell adhesion. Because loss of

E-cadherin expression results in disruption of cellular clusters, it has been postulated that E-cadherin

functions as a tumor suppressor protein. The role of E-cadherin in inflammatory breast cancer (IBC), a

distinct and highly aggressive form of breast cancer, is largely unknown. The aim of our study was to

elucidate whether E-cadherin expression contributes to the development and progression of the IBC phenotype

and to investigate any differences in E-cadherin expression between IBC and stage-matched non-IBC. Forty-two

breast cancer cases (20 IBC and 22 non-IBC) were identified. Strict and well-accepted criteria were used for

the diagnosis of IBC. Clinical and pathologic features were studied, and formalin-fixed, paraffin-embedded

tissue sections were immunostained for E-cadherin, estrogen and progesterone receptors (ER and PR,

respectively), and HER2/neu. Statistical analysis was performed using Fisher’s exact test. All IBC uniformly

expressed E-cadherin, whereas 15 of the 22 (68%) of the non-IBC expressed the protein (P = .006).

Intralymphatic tumor emboli in the IBC cases were also all E-cadherin positive. Two IBC tumors demonstrated

invasive lobular histology, and both cases were positive for E-cadherin. Of the non-IBC cases, three were

invasive lobular carcinomas, and all were positive for E-cadherin. No association was found between E-

cadherin expression and ER, PR status, or HER2/neu overexpression. Our study demonstrates that there is a

strong association between E-cadherin expression and IBC and suggests that E-cadherin may be involved in the

pathogenesis of this form of advanced breast cancer. In our study, we demonstrate that circulating IBC tumor

cells strongly express E-cadherin, thereby providing an important exception to the positive association

between E-cadherin loss and poor prognosis in breast cancer.”

2001: An intact overexpressed E-cadherin/alpha,beta-catenin axis characterizes the lymphovascular emboli of

inflammatory breast carcinoma.

Cancer Res 2001 Jul 1;61(13):5231-41, Tomlinson JS, et al.

“The step of intravasation (lymphovascular invasion), a rate-limiting step in metastasis, is greatly

exaggerated in inflammatory breast carcinoma (IBC). Comparing MARY-X with common non-IBC cell

lines/xenografts, we discovered an overexpressed and overfunctioning E-cadherin/alpha,beta-catenin axis. In

MARY-X, the E-cadherin and catenins were part of a structurally and functionally intact adhesion axis

involving the actin cytoskeleton. In vitro, MARY-X grew as round compact spheroids with a cell density 5-10

-fold higher than that of other lines. The spheroids of MARY-X completely disadhered when placed in media

containing absent Ca(2+) or anti-E-cadherin antibodies or when retrovirally transfected with a dominant-

negative E-cadherin mutant (H-2K(d)-E-cad). Anti-E-cadherin antibodies injected i.v. immunolocalized to the

pulmonary lymphovascular emboli of MARY-X and caused their dissolution. A total of 90% of human IBCs showed

increased membrane E-cadherin/alpha,beta-catenin immunoreactivity. These findings indicate that it is the

gain and not the loss of the E-cadherin axis that contributes to the IBC phenotype.”

2001: Allelic loss detection in inflammatory breast cancer: improvement with laser microdissection.

Lab Invest 2001 Oct;81(10):1397-402, Bertheau P, et al.

“SUMMARY: Solid tumors are composed not only of tumor cells but also of stromal nonneoplastic cells. In

whole tumor samples, stromal cells retaining their alleles may therefore obscure detection of loss of

heterozygosity (LOH) in tumor cells. An increasing number of studies have used laser-assisted tissue

microdissection to improve LOH detection, but the real gain in sensitivity has been poorly quantified. We

studied a group of 16 inflammatory breast carcinomas that were submitted to both standard DNA extraction

from frozen whole tumor samples and laser microdissection performed on paraffin-embedded tumor samples.

Using PCR with fluorescence-labeled primers, we comparatively analyzed ten polymorphic markers with both

sources of DNA. With the LOH detection threshold set at -25%, we showed that 25 LOHs could not be diagnosed

with whole tumor samples out of 73 LOHs positively diagnosed in microdissected samples (34%). With the LOH

detection threshold set at -50%, the respective figures were 39 LOHs not diagnosed out of 55 LOHs (71%).

Measuring the intensity of the allelic decrease, we showed that the mean decrease of the lost allele is -34%

with whole tumor samples and -67% with microdissected samples. The increase in sensitivity of LOH detection

with microdissection is associated with the density of stromal cells. This strong improvement in LOH

detection in this aggressive type of breast cancer indicates that many other molecular studies performed on

heterogeneous solid tumors may benefit from a first step of laser microdissection.”

2001: Case control study of prognostic markers and disease outcome in inflammatory carcinoma breast: a

unique clinical experience.

Breast J 2001 Nov-Dec;7(6):398-404, Aziz SA, et al.

“Inflammatory breast carcinoma (IBC) is a rare but aggressive form of breast cancer. In this first-ever

study, we investigated the role of nine prognostic markers’ expression (estrogen receptor [ER], progesterone

receptor [PR], p53, C-erbB-2, epidermal growth factor receptor [EGFR], cathepsin D [CD], proliferating cell

nuclear antigen [PCNA], DNA ploidy, and S-phase fraction [SPF]) and disease outcome in IBC cases compared

with the control group. A case control study of IBC was conducted on 40 test cases with two controls per

case matching age, grade, and number of axillary lymph nodes sampled. During 7 years of this study, 10% of

all patients with breast cancer had IBC. In this study, 84% of IBC cases showed positive axillary lymph

nodes compared with 63% in control group. The expression of nine prognostic markers, that is, ER, PR, p53,

C-erbB-2, EGFR, CD, PCNA, SPF, and DNA ploidy, was studied by immunohistochemistry and flow cytometry.

Hormone receptor status showed an inverse correlation (p < 0.05). Among p53, C-erbB-2, EGFR, and CD in the

IBC group, only p53 showed a significant correlation, with 70% positivity in IBC versus 48% positivity in

the control group (p < 0.05). Much higher SPF and PCNA positivity was seen in the IBC group compared with

the control group (p < 0.05). DNA ploidy also showed a significant correlation compared with the control

group (p < 0.05). After a median follow up of 18 months, median overall survival in the IBC group was 1.8

years (range 0.6-5.8 years) compared with 3.0 years (range 2.5-7.0 years), with a p value of 0.0001.”

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2002: Hemodynamics in vasculogenic mimicry and angiogenesis of inflammatory breast cancer xenograft.

Cancer Res 2002 Jan 15;62(2):560-6, Shirakawa K, et al.

“In the present study, we examined hemodynamics in vasculogenic mimicry (VM) and angiogenesis of

inflammatory breast cancer (IBC) xenografts (WIBC-9), having previously reported on the unique histological

features and molecular basis of these processes (K. Shirakawa et al., Cancer Res., 61: 445-451, 2001).

Histologically, the WIBC-9 xenografts exhibited invasive ductal carcinoma with a hypervascular structure

(angiogenesis) in the tumor margin and VM without endothelial cells, central necrosis, or fibrosis in the

tumor center. Results of molecular analysis indicated that WIBC-9 had a vasculogenic phenotype, including

expression of Flt-1 and Tie-2. Comparison of WIBC-9 with an established non-IBC xenograft (MC-5), using

time-coursed dynamic micromagnetic resonance angiography analysis (with our newly developed intravascular

macromolecular magnetic resonance imaging contrast agent), electromicroscopy, and immunohistochemistry,

demonstrated blood flow and a VM-angiogenesis junction in the central area of the WIBC-9 tumor. It has

previously been considered impossible to prove a connection between VM and angiogenesis using angiography,

because there are no intravascular macromolecular magnetic resonance imaging contrast agents that do not

exhibit significant leakage through the vascular wall. In the present study, laser-captured microdissection

was performed in regions of WIBC-9 tumors that exhibited VM without endothelial cells, central necrosis, or

fibrosis, revealing expression of human-Flt-1 and human-Tie2 and the absence of human-CD31, human-endothelin

B receptor, and human-thrombin receptor. These facts led us to hypothesize that the VM of WIBC-9 involves

hemodynamics that serve to feed WIBC-9 cells, and this in turn suggests a connection between VM and

angiogenesis.”

2002: Rapid Accumulation and Internalization of Radiolabeled Herceptin in an Inflammatory Breast Cancer

Xenograft with Vasculogenic Mimicry Predicted by the Contrast-enhanced Dynamic MRI with the Macromolecular

Contrast Agent G6-(1B4M-Gd)(256).

Cancer Res 2002 Feb 1;62(3):860-6, Kobayashi H, et al.

“The rapid blood flow and perfusion of macromolecules in the inflammatory breast cancer xenograft (WIBC-9),

which exhibits a “vasculogenic mimicry” type of angiogenesis without the participation of endothelial cells

and expresses high levels of the HER-2/neu antigen, was evaluated in mice using 3D-micro-MR angiography

using a novel macromolecular MR contrast agent [G6-(1B4M-Gd)(256)]. Herceptin, which recognizes the HER-

2/neu antigen and has similar size (10 nm) to G6-(1B4M-Gd)(256), accumulated and internalized in the WIBC-9

tumors more quickly than in the control MC-5 tumors that progress with normal angiogenesis. Three

dimensional micro-MRI with the G6-(1B4M-Gd)(256) macromolecular MRI contrast agent distinguishes between the

different types of angiogenesis and is predictive of the rapid accumulation and internalization of Herceptin

in the WIBC-9 inflammatory breast cancer xenograft.”

2002: The Molecular Basis of Inflammatory Breast Cancer.

California Breast Cancer Research Program Symposium, Barsky SH, et al.

“Introduction: Inflammatory breast cancer (IBC) is a poorly understood, little studied form of breast cancer

which is very aggressive and particularly devastating in disadvantaged minority women. IBC is characterized

by florid tumor emboli within lymphovascular spaces, a phenotype which distinguishes it from other forms of

breast cancer. The molecular basis of this phenotype is the focus of this research.

“Methods: Using a novel human-scid model of IBC, we have conceptually analyzed this phenotype in three

parts.

“Results:

1. The tumor cell embolus (IBC spheroid) forms on the basis of an intact and overexpressed E-cadherin /

alpha-, beta-catenin axis which mediates tumor cell-tumor cell adhesion analogous to the embryonic

blastocyst and accounts for both the compactness of the embolus and its complete dissolution with anti-E-

cadherin antibodies, absent calcium, or E-cadherin dominant-negative mutant approaches. Dissolution of the

tumor cell embolus by any of these approaches induces apoptosis via an anoikis pathway. The compactness of

the embolus results in its resistance to chemotherapy / radiation therapy and its efficiency at metastasis

formation and therefore therapeutic strategies which disadhere it are highly desirable.
2. The tumor cell embolus (IBC spheroid), in contrast, fails to bind the surrounding vascular endothelial

cells because of complete absence of sialyl-Lewis X/A carbohydrate ligand-binding epitopes on its

overexpressed MUC1 which are necessary for binding endothelial cell E-selectin. This natural tumor cell-

endothelial cell aversion of the tumor cell embolus (IBC spheroid) further contributes to the compactness of

the IBC spheroid and its passivity in metastasis dissemination. Experiments with purified

glycosyltransferases which add sialyl-Lewis X/A to MUC1 on the IBC spheroids produce strong electrostatic

repulsions which disrupt the E-cadherin homodimers and cause disadherence.
3. The tumor cell embolus finds itself within the vascular lumen in the first place because it stimulates

a vascular channel to form around it rather than intravasating into a pre-existing lymphatic or capillary.

The enveloping vascular channel does not form from angiogenesis but rather from vasculogenesis as evidenced

by experiments where tumor cell emboli (IBC spheroids) are admixed with murine embryonal fibroblasts labeled

with green fluorescent protein (GFP) and injected into scid mice. Tumor emboli are observed within

lymphovascular spaces where the endothelial cells express vasculogenesis markers as well as endothelial

markers. These endothelial cells also express GFP, evidence that they must have formed from the injected

GFP-labeled murine embryonal fibroblasts.

“Conclusion: The molecular basis of IBC provides a mechanism by which IBC bypasses the traditional steps of

intravasation, dissemination and extravasation in its metastatic pathway and affords opportunities for

therapeutic intervention.”