The value of progesterone receptor expression in predicting the Recurrence Score for hormone-receptor positive invasive breast cancer patients.

BACKGROUND: OncotypeDX(®) (ODX) is a well-validated assay for breast cancer treatment planning. We explored whether the conventional pathological factors could pick up high risk patients without the help of the ODX. METHODS: The ODX was performed on 139 hormone receptor-positive invasive breast cancers in a single Japanese institution. The recurrence risk was compared between the ODX and the St. Gallen Consensuses. The correlations were analyzed between the Recurrence Score (RS) measured by ODX and the pathological factors. In addition, we performed a follow-up survey and examined the association of the RS with the confirmed recurrence or death. RESULTS: The ODX classified 68 (49 %) as low RS, 52 (37 %) as intermediate RS, and 19 (14 %) as high RS cases. Correlations were noted between RS and progesterone receptor (PR) (r = -0.53), Ki-67 (r = 0.42), and nuclear grade (NG) (r = 0.41). None had a high RS with PR(3+) or NG1. Only one high RS patient had a Ki-67 (<20 %). The combinations of high RS with PR(0)/Ki-67 (≥20 %) and PR(1+)/Ki-67 (≥20 %) were 70 and 58 %, respectively. The combinations with high RS and PR(0)/NG3, PR(0)/NG2, and PR(1+)/NG3 were 83, 75, and 75 %, respectively. The median follow-up was 39.1 months (range 24.0-67.8). There were one low RS (1 %), four intermediate RS (8 %), and three high RS patients (16 %) who developed local or distant recurrence. CONCLUSION: Hormone receptor-positive invasive breast cancers are stratified with the combinations of PR/Ki-67 or PR/NG. Some of the high recurrence risk cases might be identified without the ODX.

Ultrasonography mapping combined with mammography before breast-conserving surgery for primary breast cancer with microcalcifications: a novel approach.

INTRODUCTION: Evaluation for the spread of breast cancer with microcalcifications is challenging, because the microcalcifications sometimes spread beyond the lesions detectable by ultrasonography (US). An original method for preoperative mapping was performed for such lesions, using US in combination with mammography (MG) (US + MG mapping) before breast-conserving surgery (BCS). MATERIALS AND METHODS: A total of 885 consecutive patients underwent BCS for primary breast cancer. Of the 885 patients, 154 (17.4%) with ductal carcinoma in situ or invasive carcinoma having microcalcifications underwent US + MG mapping preoperatively. Five patients who received neoadjuvant chemotherapy and 17 patients who were lost to follow-up were excluded. Accordingly, 133 lesions in 132 patients were retrospectively evaluated. The associations among this method, surgical margin (positive, close, or negative), pathologic characteristics, the area of the lesion within the specimen, and local recurrence rate during 5 years of follow-up were analyzed. RESULTS: The median age and follow-up duration were 51.3 years (range, 28-80 years) and 71.4 months (range, 60-79 months), respectively. The surgical margin was negative in 96 lesions (72.2%), close in 27 lesions (20.3%), and positive in 10 lesions (7.5%). Local recurrence was noted in 1 patient (0.8%). There was no significant association between surgical margin status and the presence of invasive carcinoma. Larger lesion area was significantly associated with positive or close margin (P = .027). CONCLUSION: US + MG mapping is useful and results in a high complete-resection rate and an extremely low 5-year local recurrence rate.

FR-167653, a selective p38 MAPK inhibitor, exerts salutary effect on liver cirrhosis through downregulation of Runx2.

Liver cirrhosis remains a difficult-to-treat disease with a substantial morbidity and mortality rate. There is an emerging body of data purporting a pivotal role of the activated p38 mitogen-activated protein kinase (MAPK) in the process of cirrhosis. Several anticirrhotic agents have been developed over the past few years, and most of them exert their effects by indirectly inhibiting the p38 pathway. Effect of a selective p38 inhibitor is yet to be reported. In this study, we evaluated the salutary effect of FR-167653 (FR), a selective p38 inhibitor, in a carbon tetrachloride (CCl(4))-induced rat cirrhotic model. Twenty rats were assigned into four groups: Sham, olive oil only; Control, CCl(4) in olive oil; FR50, FR 50 mg/kg/day and CCl(4); and FR100, FR 100 mg/kg/day and CCl(4). FR dose-dependently inhibited activation of p38 and had an ameliorating effect on cirrhosis formation. Significant dose-dependent reduction in alpha-smooth muscle actin immunostaining and hydroxyproline content of the liver was noticed in the FR-treated rats. Also densitometric analysis showed a significant reduction in azan-stained area in the FR-treated rats. These fibrotic changes were observed in the myofibroblasts including the hepatic stellate cells and portal fibroblasts. mRNA expression of runt-related protein 2 (Runx2), a profibrogenic transcription factor, was significantly low in FR-treated livers, indicating that Runx2 might be a key downstream regulator of the p38 pathway. A similar reduction in expression of Smad4 and tissue inhibitor of metalloproteinase-1 was noticed in the FR-treated rats. In conclusion, FR treatment exerted a significant beneficial effect in a CCl(4)-induced rat cirrhotic model. The ameliorating effect of FR could be partially attributable to an inhibition of the Smad4/p38/Runx2 axis in the cirrhotic liver.

Differential expression of RUNX genes in human esophageal squamous cell carcinoma: downregulation of RUNX3 worsens patient prognosis.

BACKGROUND: The RUNX proteins are a family of transcriptional factors that have essential functions during embryogenesis and development, whereas deregulation in expression of RUNXs is often linked to tumor formation. To date, there has been no study describing the precise expression, prognostic impact and methylation status of RUNXs in esophageal squamous cell carcinoma. METHODS: Resected specimens from 61 patients with esophageal SCC were used to identify the expression of RUNX1, RUNX2 and RUNX3 by real-time RT-PCR. Localization of mRNA was done by in situ hybridization, expression of Smad4 was evaluated by immunohistochemistry, and the methylation status of RUNX3 was analyzed by methylation-specific PCR. RESULTS: RUNX3 had a significant impact on patient prognosis with worse survival in the RUNX3-negative group. In early tumors (T1/T2), the prevalence of lymph vessel invasion and the number of metastatic lymph nodes were significantly higher in RUNX3-negative tumors. Furthermore, RUNX3 became a strong prognostic factor only in Smad4-positive tumors. Also, the methylation status of the RUNX3 promoter had a significant correlation with the loss of RUNX3 expression. CONCLUSION: Downregulation of RUNX3 may play a role in disease progression of esophageal SCC, and hypermethylation of the promoter region might be one of the crucial pathways to silence RUNX3 gene.

Tetracycline analogues (doxycycline and COL-3) induce caspase-dependent and -independent apoptosis in human colon cancer cells.

Tetracycline analogues (TCNAs) possess cytotoxic activities as well as matrix metalloproteinase (MMP) inhibitory properties. Previously, we demonstrated that doxycycline (DOXY) could induce apoptosis in human HT29 colon cancer cells. In present study, the molecular apoptotic mechanisms induced by two kinds of TCNAs, designated as DOXY and COL-3 (chemically modified tetracycline-3; 6-demethyl, 6-deoxy, 4-dedimethylamino tetracycline), were evaluated in cultured HT29 cells. Both TCNAs inhibited the proliferation of 6 different colorectal cancer cell lines in a dose-dependent manner. Especially, COL-3 had a stronger effect on cancer cells than DOXY. Apoptotic changes were actually observed by 10 mug/ml COL-3 and 20 mug/ml DOXY in a time-dependent manner. COL-3 produced the increase in cytosolic cytochrome c and the loss of mitochondrial membrane potential after 3 hr treatment, and thereafter activated caspases. In case of DOXY, these changes were observed after 24 hr. Bax translocation was not a prerequisite for cytochrome c releasing in COL-3 treatment. Pretreated pancaspase inhibitor (Z-VAD-FMK) reduced COL-3 and DOXY mediated apoptosis up to 81.3 and 35.3%, as compared with nontreated cells, respectively. These data indicated that TCNAs could induce mitochondria-mediated apoptosis through both caspase-dependent and -independent pathway. In fact, endonuclease G and apoptosis-inducing factor were released into cytosol after the treatment of TCNAs, which indicated that caspase-independent apoptotic pathway is also one of the key mechanisms for the treatment of TCNAs. Taken together, we believe that TCNAs could have strong potentials for clinical application in treating colorectal cancers and improve cancer chemotherapy.

Doxycycline inhibits cell proliferation and invasive potential: combination therapy with cyclooxygenase-2 inhibitor in human colorectal cancer cells.

Matrix metalloproteinases (MMPs) and cyclooxygenase (COX) enzymes play pivotal roles in the metastatic process of colorectal cancers. Inhibition of both MMPs and COX could be an attractive option for the inhibition of cell growth and invasion. Two human colorectal cancer cell lines, LS174T and HT29, were challenged with MMP inhibitor (doxycycline), selective COX-2 inhibitor (NS-398), or a combination of these agents to evaluate cancer cell proliferation and invasion. Dose-dependent growth inhibition was observed in both cell lines when they were treated with a single therapy. These effects were not related to MMP-2 or MMP-9 expression potential of the cell lines. Doxycycline (10 microg/mL) induced G(0)/G(1) arrest, and 20 microg/mL provoked annexin V positivity and up-regulated caspase-3 activity in HT29 cells. However, 20 microg/mL doxycycline caused no distinct apoptotic change in LS174T cells. Although MMP expression was not inhibited by 5 to 10 microg/mL doxycycline or 50 to 100 micromol/L NS-398, MMPs' activities were down-regulated by these concentrations. Cellular invasion was noticed in LS174T cells, but their capacity for invasion was diminished by these inhibitors. The antiproliferative and antiinvasive effects of the combination therapy were more pronounced. Doxycycline (5 microg/mL) with 50 micromol/L NS-398 inhibited cell proliferation and doxycycline (5 microg/mL) with 100 micromol/L NS-398 attenuated MMP expression and activity, as well as capacity for invasion, compared with single therapy. These data suggest that combination therapy consisting of an MMP inhibitor with a COX-2 inhibitor is an attractive approach to the treatment of colorectal cancers. The use of this treatment regimen for chemoprevention or treatment of colorectal cancers should be considered in future clinical trials.