Evidence for coordinated interaction of cyclin D3 with p21 and cdk6 in directing the development of uterine stromal cell decidualization and polyploidy during implantation.

Uterine decidualization, characterized by stromal cell proliferation, and differentiation into specialized type of cells (decidual cells) with polyploidy, during implantation is critical to the pregnancy establishment in mice. The mechanisms by which the cell cycle events govern these processes are poorly understood. The cell cycle is tightly regulated at two particular checkpoints, G1-S and G2-M phases. Normal operation of these phases involves a complex interplay of cyclins, cyclin-dependent kinases (cdks) and cdk inhibitors (CKIs). We previously observed that upregulation of uterine cyclin D3 at the implantation site is tightly associated with decidualization in mice. To better understand the role of cyclin D3 in this process, we examined cell-specific expression and associated interactions of several cell cycle regulators (cyclins, cdks and CKIs) specific to different phases of the cell cycle during decidualization in mice. Among the various cell cycle molecules examined, coordinate expression and functional association of cyclin D3 with cdk4 suggest a role for proliferation and, that of cyclin D3 with p21 and cdk6 is consistent with the development of polyploidy during stromal cell decidualization.

Transgelin: a potentially useful diagnostic marker differentially expressed in triple-negative and non-triple-negative breast cancers.

Triple negative (TN) (estrogen receptor [ER], progesterone receptor [PR] and HER2-) are highly aggressive, rapidly growing, hormone-unresponsive tumors diagnosed at later stage that affect younger women with shorter overall survival. Most TN tumors are of the basal type. For the remainder, identification of target markers for effective treatment strategies remains a challenge. Transgelin (TGLN) is a 22-kd actin-binding protein of the calponin family. It is one of the earliest markers of smooth muscle differentiation. TGLN has been shown to have important biologic activities including regulating muscle fiber contractility, cell migration, and tumor suppression. We examined TGLN expression in the different molecular subtypes of breast cancer. TGLN expression was examined as a function of tumor size, grade, histologic type, lymph node status, patients' age and overall survival, ER, PR, HER2, and Ki-67 in 101 tumors that included 35 luminal A, 28 luminal B, 4 HER2, and 34 TN types. TGLN positivity (defined as 2+ or 3+) was associated with more aggressive tumors (10% of grade I/II tumors were TGLN+ versus 53% of grade III tumors; P < .001), high Ki-67 count, and low ER and PR expression (P < .001) but not with tumor size, age, or lymph node metastasis. TN (n = 34) tumors were 7.7 times more likely to be TGLN+ than non-TN (n = 67) tumors (77% versus 10%, respectively; P < .001). TGLN may be an excellent diagnostic marker of TN tumors and could be useful in stratification of patients. TGLN may also prove a potential target for future treatment strategies.

A comparison of prognostic tumor markers obtained on image-guided breast biopsies and final surgical specimens.

BACKGROUND: This study was initiated to determine whether tumor markers obtained on image-guided breast biopsy specimens provide accurate prognostic information for women with invasive breast cancer. METHODS: Prognostic tumor markers on preoperative image-guided biopsy and final surgical specimens were compared in 44 patients with invasive breast cancer. RESULTS: Progesterone receptor (PR) discordance was 18%. In 87% of PR discordant cases, the image-guided biopsy was positive and the final specimen was negative (P = 0.03). Tumor grade was discordant in 36% of patients Discordance for estrogen receptor (ER) = 2%; MIB-1 = 18%; Her2/neu = 9%; EGFR = 10%; p53 = 9%; and bcl-2 = 0%. The discordance for these markers was random and did not reach statistical significance. CONCLUSION: Image-guided core needle biopsies provide reliable information for the majority of prognostic tumor makers. A positive progesterone receptor is significantly more likely to be determined by core biopsy rather than the final surgical specimen. Tumor grade should be based upon the final surgical specimen whenever possible.

Ki-67 expression in axillary lymph node metastases in breast cancer is prognostically significant.

Several studies have documented the prognostic significance of cell proliferation in breast cancer and its positive relationship with tumor grade, size, mitotic activity, hormonal and Her-2 status, and tumor progression. The Ki-67 antigen provides an accurate measure of the growth fraction of a tumor. Ki-67 expression in 103 primary breast carcinomas and their corresponding axillary lymph node metastases was correlated with age, tumor grade, size, estrogen receptor (ER), progesterone receptor (PgR), p53, epidermal growth factor receptor (EGFR), Bcl-2, Her-2 status, and patients' overall survival. Median Ki-67 expression in primary and metastatic tumors was 20% and 15%, respectively. Although there was no difference in overall survival (P = .65, log-rank test) between primary tumors with less than or at least 10% Ki-67 expression, there was significantly better overall survival when Ki-67 expression in lymph nodes was less than 10% (P = .040). For patients whose primary tumors exhibited Ki-67 expression less than 10%, most of their metastatic lesions had a similar low Ki-67; these patients had a favorable outcome. A small subgroup was noted to have a nodal Ki-67 of 10% or more and worse survival (P = .047). For patients whose primary tumors had a Ki-67 of 10% or more, most of their metastatic lesions had similar high Ki-67 values; however, a group of 12 patients had lymph node Ki-67 less than 10% and had a better overall survival (P = .092). Our results showed that measurement of Ki-67 in lymph node is superior to its evaluation in primary tumors. Identification of subgroups of patients in whom Ki-67 expression in lymph nodes differs from expression in primary tumor may assist in the selection of therapeutic options.

Prognostic significance of Bcl-2 in invasive mammary carcinomas: a comparative clinicopathologic study between "triple-negative" and non-"triple-negative" tumors.

Bcl-2 is a tumorigenic protein that is expressed in 25% to 50% of breast cancers. Although its expression has been widely accepted as a favorable prognostic marker, its protective mechanism of action remains unclear. "Triple-negative" tumors are an aggressive subgroup known to carry a poor prognosis. Studies documenting prognostic significance of Bcl-2 expression in triple-negative in comparison to non-triple-negative breast cancers are limited. Bcl-2 expression was correlated with tumor size, grade, histologic type, lymphovascular invasion, lymph node status, patients' overall survival, estrogen receptor, progesterone receptor, Her-2, p53, and epidermal growth factor receptor in 124 triple-negative and 458 non-triple-negative tumors. There were significant differences between triple-negative and non-triple-negative tumors in their relationship to Bcl-2 expression (81% versus 29%, respectively) and tumor aggression. As previously reported, in non-triple-negative tumors, Bcl-2 positivity correlated with less aggressive tumors (94% of grade I tumors were Bcl-2+ versus 62% of grade III tumors, P < .011) and overall survival (P = .008). However, the opposite was true in patients with triple-negative tumors, where Bcl-2 positivity was associated with poorer survival (P = .64). In triple-negative tumors, Bcl-2 positivity was not associated with any of the aforementioned parameters except for a lower incidence of lymph node metastasis. Moreover, by Cox regression analysis of all variables, in patients with triple-negative tumors, lymphovascular invasion (P = .009) and Bcl-2 expression (P = .028) were predictors of poor survival. In conclusion, there are major clinicopathologic differences between breast cancer phenotypes. Our results establish the value of using Bcl-2 in prognostic stratification of patients and its potential therapeutic implications in selecting patients for treatment.

Clinicopathological characteristics of triple-negative invasive mammary carcinomas in African-American versus Caucasian women.

African-American (AA) women are more likely to have late stage, aggressive, rapidly growing, and less hormone-responsive breast tumors. An aggressive subtype of cancer, known as "Triple-Negative" (TN), that is negative for Her-2 and for estrogen and progesterone receptors (ER and PR), is reported to be more common in AA women. We examined the clinical, histopathologic, and prognostic features of TN tumors in AA and Caucasian women. Tumor size, grade, histologic type, lymphovascular invasion (LVI), lymph node status, patient survival, ploidy status, and expression of ER, PR, p53, epidermal growth factor receptor (EGFR), MIB-1, Bcl-2, Her-2, p27, and p21 were evaluated. The TN tumors (75%) were high grade, large, aneuploid tumors that occurred in younger women and were more likely to have a high rate of LVI, elevated MIB-1 score, and nodal metastases. Patients with TN tumors showed poorer overall survival. There was no difference in overall or disease-free survival (p = 0.46) in the AA versus Caucasian women. LVI was a significant predictor of overall survival in AA but not in Caucasian women. There were minor differences in histopathologic features, biomarker expressions, and survival in AA and Caucasian women with TN tumors. The absence of LVI in AA patients predicted an excellent probability of survival.

A newly proposed semi-automated method of grading invasive lobular carcinoma: a unifying concept and correlation with prognostic markers and patient survival.

Invasive ductal carcinoma (IDC) of the breast is currently graded according to the Nottingham modification of the Scarff-Bloom-Richardson system (SBR). This system involves subjective evaluation of 3 morphologic features: tubule formation, nuclear pleomorphism, and mitosis. Our recently proposed semi-automated Nuclear and Proliferation Index [N+P] grading system for IDC has demonstrated agreement among grades and prognostic markers with better prediction of patient survival than the SBR system. Our present objective is to expand the utilization of the N+P system to grading invasive lobular carcinoma (ILC). Fifty-eight ILC cases were evaluated by the SBR and N+P systems. The 2 systems were compared in terms of correlation with patient survival, tumor size, grade, angiolymphatic invasion, lymph node status, ploidy status, and ER, PR, Her-2, p53, EGFR, and Bcl-2 staining. The N+P and SBR systems demonstrated overall agreement when correlated with clinical and prognostic parameters. Twenty-four of 30 tumors initially classified as SBR Grade II were down-graded to N+P I. Three of 26 tumors initially classified as SBR Grade I were up-graded to N+P II. Grading of ILC provides valuable predictive and prognostic information. The N+P grading system for ILC decreases the element of subjectivity for assessing mitotic activity and appears to be superior to the SBR system in predicting patient survival.

Biphasic acute myeloid leukemia with near-tetraploidy and immunophenotypic transformation.

This report describes a case of acute myeloid leukemia (subtype M1) with biphasic morphology. The bone marrow biopsy showed 2 distinct regions of blasts, one containing large cells and the other small cells. Morphometric and DNA ploidy analysis showed that the mean nuclear area and mean DNA index for the large cell region were 2-fold higher than those for the small cell region. Cytogenetic analysis showed an abnormal near-tetraploid clone. The tumor relapsed following aggressive therapy. The cells from the relapse specimen were similar to the original small cell region with respect to nuclear area and DNA index; however, there was immunophenotypic transformation with gain of CD7 and gain of CD56. Cytogenetically, the relapse specimen showed no evidence of the near-tetraploid clone, but instead had a previously unidentified abnormal clone containing 46 chromosomes and structural abnormalities of 2q and 7q. Biphasic morphology in acute myeloid leukemia may be predictive of a near-tetraploid subclone and immunophenotypic transformation.

HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation.

Stromal cell polyploidy is a unique phenomenon that occurs during uterine decidualization following embryo implantation, although the developmental mechanism still remains elusive. The general consensus is that the aberrant expression and altered functional activity of cell cycle regulatory molecules at two particular checkpoints G1 to S and G2 to M in the cell cycle play an important role in the development of cellular polyploidy. Despite the compelling evidence of intrinsic cell cycle alteration, it has been implicated that the development of cellular polyploidy may be controlled by specific actions of extracellular growth regulators. Here we show a novel role for heparin-binding EGF-like growth factor (HB-EGF) in the developmental process of stromal cell polyploidy in mice. HB-EGF, which is one of the earliest known molecular mediators of implantation in mice and humans, promotes stromal cell polyploidy via upregulation of cyclin D3. Adenoviral delivery of antisense cyclin D3 attenuates cyclin D3 expression and abrogates HB-EGF-induced stromal cell polyploidy in vitro and in vivo. Collectively, the results demonstrate that the regulation of stromal cell polyploidy and decidualization induced by HB-EGF depend on cyclin D3 induction.