Flow cytometry and quantitative immunohistochemical study of cell cycle regulation proteins in invasive breast carcinoma

Abstract

BACKGROUNDBetween January 11, 1991 and January 8, 1992, 104 patients with previously untreated, invasive, primitive breast carcinoma were admitted to the authors' hospital.METHODSFor each patient, flow cytometry DNA analyses on frozen samples and on immunohistochemical staining were performed, including Ki‐67, cyclin A, p53, and p21waf1 (p21), with assessment of the percentages of positive nuclei were assessed. Correlations with classic clinicopathologic data and survival (overall, metastasis free, or recurrence free) and a multivariate analysis were performed.RESULTSAfter a multivariate analysis according to a Cox model that was stratified by age, tumor size, tumor grade, lymph node status, and receptor status, among the factors studied, the presence of p21 was the unique remaining prognostic factor for patients with invasive breast carcinoma. Because of the lack of a correlation between p21 and proliferative factors (Ki‐67, S‐phase, and cyclin A), the authors combined p21 with those markers and found that, for the different combinations, after statistical analysis, only p21 combined with S‐phase or with cyclin A and lymph node status were salient survival prognostic factors.CONCLUSIONSImmunohistochemical study of proteins involved in the cell cycle and assessment of proliferative activity using flow cytometric DNA analysis aided the authors in singling out correlations of cyclin A and S‐phase, along with p21, with metastasis free survival and overall survival in patients with invasive breast carcinoma. These promising results will require confirmation in a larger series of patients. Cancer 2003;97:1376–86. © 2003 American Cancer Society.DOI 10.1002/cncr.11209

Cyclin A as a marker for prognosis and chemotherapy response in advanced breast cancer

“…We found a strong correlation between cyclin A, mitotic count, tumour grade and Ki-67, which is in line with previous studies in breast cancer (Michalides et al, 2002;Michels et al, 2002). Cyclin A was the only marker that showed a statistically significant correlation to both TFR and OS; therefore, it seems to be the most useful marker of proliferation.…”

“…We used a cutoff value of 10.5% for the high and low cyclin A positivity groups. This is in line with three other studies, where median values varying between 8 and 11% have been used (Michalides et al, 2002;Michels et al, 2002;Kuhling et al, 2003;Rudolph et al, 2003). In the fourth study, four different categories were used for cyclin A positivity and the median category was between 15 and 30% (Bukholm et al, 2001).…”

“…An association between a high cyclin A expression and poor prognosis has previously been described in three studies, all executed after primary therapy for breast cancer (Bukholm et al, 2001;Michalides et al, 2002;Michels et al, 2002). A fourth study, however, in node-negative patients showed no correlation between cyclin A and prognosis Rudolph et al, 2003).…”

“…Cyclin A is essential in the S phase for DNA replication, it is also involved in G2 -M-phase transition (Zindy et al, 1992;Fan and Bertino, 1997), and is therefore one of the most useful markers signifying proliferating cells. Overexpression of cyclin A has been associated with worse prognosis for breast cancer in several studies (Bukholm et al, 2001;Michalides et al, 2002;Michels et al, 2002), but the results have not been consistent Rudolph et al, 2003). Since the lack of prognostic significance of cyclin A is mainly seen in patients with lymph node-negative breast cancer, it is possible that cyclin A plays a more important role in aggressive tumours and metastatic disease .…”

Ki-67 as prognostic marker in early breast cancer: a meta-analysis of published studies involving 12 155 patients

“…HR associated with Ki-67 positivity in this case) from studies with inherent dissimilarities. The observed disparity among the conclusions of different studies, responsible for the observed heterogeneity, can be quantified by applying Bukholm et al (2003) 147 No Ceccarelli et al (2000) 217 Yes Galiegue et al (2004) 117 No Gasparini et al (1992) 164 Yes Haerslev et al (1996) 487 Yes Jalava et al (2000) 414 No Kroger et al (2006) 157 No Kronblad et al (2006) 377 Yes Lampe et al (1998) 142 Yes Liu et al (2000) 225 Yes Michels et al (2003) 104 No Molino et al (1997) 322 Yes Rudas et al (1994) 184 No Tsutsui et al (2005) 249 Yes Yang et al (2003) 147 No…”