CDKN2A inhibits formation of homotypic cell-in-cell structures.

Cell-in-cell (CIC) structures, characterized by enclosure of one or more cells within another cell, were extensively documented in human cancers. Although elevated CIC formation was found in cancers with CDKN2A inactivation, a causal link between them remains to be established. We reported here that inhibiting CDKN2A expression effectively promoted homotypic CIC formation, whereas ectopic overexpression of p16INK4a or p14ARF, two proteins encoded by CDKN2A gene, significantly suppressed CIC formation in MCF7 cells. The regulation of CIC formation by CDKN2A was tightly correlated with subcellular redistribution of E-cadherin, F-actin rearrangement and reduced phosphorylation of myosin light chain 2 (p-MLC2), consistent with which, CDKN2A expression imparted cells winner/outer identity in competition assay. Moreover, CIC formation negatively correlates with p16INK4a expression in human breast cancers. Thus, our work identifies CDKN2A as the first tumor suppressor whose inactivation promotes homotypic CIC formation in human cancer cells.

A nomogram for predicting survival in patients with breast cancer brain metastasis.

Brain metastasis (BM) is common in patients with breast cancer. Predicting patient survival is critical for the clinical management of breast cancer brain metastasis (BCBM). The present study was designed to develop and evaluate a prognostic model for patients with newly diagnosed BCBM. Based on the clinical data of patients with BCBM treated in the Affiliated Hospital of Academy of Military Medical Sciences (Beijing, China) between 2002 and 2014, a nomogram was developed to predict survival using proportional hazards regression analysis. The model was validated internally by bootstrapping, and the concordance index (c-index) was calculated. A calibration curve and c-index were used to evaluate discriminatory and predictive ability, in order to compare the nomogram with widely used models, including recursive partitioning analysis (RPA), graded prognostic assessment (GPA) and breast-graded prognostic assessment (Breast-GPA). A total of 411 patients with BCBM were included in the development of this predictive model. The median overall survival time was 14.1 months. Statistically significant predictors for patient survival included biological subtype, Karnofsky performance score, leptomeningeal metastasis, extracranial metastasis, the number of brain metastases and disease-free survival. A nomogram for predicting 1- and 2-year overall survival rates was constructed, which exhibited good accuracy in predicting overall survival with a concordance index of 0.735. This model outperformed RPA, GPA and Breast-GPA, based on the comparisons of the c-indexes. The nomogram constructed based on a multiple factor analysis was able to more accurately predict the individual survival probability of patients with BCBM, compared with existing models.

Circulating plasma microRNAs as potential markers to identify EGFR mutation status and to monitor epidermal growth factor receptor-tyrosine kinase inhibitor treatment in patients with advanced non-small cell lung cancer.

We aimed to identify a panel of circulating plasma microRNAs that can predict EGFR mutation status and monitor epidermal growth factor receptor-tyrosine kinase inhibitor treatment in patients with non-small cell lung cancer. Microarrays were performed for the preliminary screening of dysregulated microRNAs in 9 EGFR mutation-positive patients versus healthy controls. MiR-107 was upregulated and miR-195 was downregulated in the exon 19 deletion versus wild-type group. The areas under the receiver operating characteristic curves for miR-107, miR-195, and a panel of these 2 microRNAs were 0.72, 0.75, and 0.74, with sensitivities and specificities of 64.7% and 76.6%, 71.8% and 69.1%, and 71.7% and 78.9%, respectively. MiR-122 was significantly upregulated in the p.L858R versus wild-type group. An area under the receiver operative characteristic curve of 0.75 suggests that miR-122 might be a specific biomarker for patients with the p.L858R mutation. In addition, dynamic changes in these 3 microRNAs were also found to correlate with responses to epidermal growth factor receptor-tyrosine kinase inhibitor treatment, indicating that circulating plasma microRNAs may represent potential biomarkers for monitoring epidermal growth factor receptor-tyrosine kinase inhibitor treatment. This study demonstrates the prospective application of circulating plasma microRNAs as potential non-invasive, convenient biomarkers for patients with EGFR-sensitive mutations.

Bone metabolism markers: Indicators of loading dose intravenous ibandronate treatment for bone metastases from breast cancer.

To investigate the changes in bone metabolism markers after second-line treatment with loading dose intravenous (i.v.) ibandronate in patients with bone metastases (BM) from breast cancer, 80 patients were enrolled in this study during January 2010 to April 2014. All the patients were treated with a second-line loading dose ibandronate for advanced breast cancer with BM and moderate-to-severe bone pain. Ibandronate (6 mg) was intravenously administered on three consecutive days followed by maintenance treatment every 3-4 weeks. Clinical data, including pain score, Karnofsky performance status (KPS) score, and changes in bone metabolism markers, were analyzed. Sixty-two patients were included in the final analysis. Compared with their pre-treatment scores, patients exhibited significantly increased KPS scores (P < .01) and a reduced dose of analgesic medication (oxycodone) (P < .01) after 3 and 6 weeks' post-treatment. The levels of serum bone alkaline phosphatase (BAP), tartrate-resistant acid phosphatase (TRACP-5b), and cross-linked carboxy-terminal telopeptide of type I collagen (ICTP) were significantly reduced after 3 and 6 weeks' post-treatment (P < .001). Aside from a few adverse events, no liver or renal toxicity was observed. Bone metabolism markers decreased by varying degrees after treatment with a loading dose of ibandronate in patients with BM from breast cancer. It might be convenient using bone metabolism markers to potentially evaluate the efficacy of bisphosphonates treatment for bone metastasis.

Receptor conversion in metastatic breast cancer: a prognosticator of survival.

OBJECTIVE: This retrospective study investigated the association between hormone receptor (HR) conversion and survival in breast cancer patients. METHODS: Estrogen receptor (ER) and progesterone receptor (PR) status (positive or negative) of primary tumors and of paired metastatic sites in 627 breast cancer patients were analyzed by McNemar's test for rates of receptor conversion. A survival analysis was performed using the Kaplan-Meier method, and prognostic factors were assessed using Cox's proportional hazards regression model. RESULTS: Conversion of ER occurred in 165 (26.31%) patients, and conversion of PR in 213 (33.97%; P < 0.001, both). For 82 patients whose ER and PR were reassessed 2-4 times during metastatic progression, ER and PR re-conversion occurred in 22 (26.83%) and 29 (35.36%), respectively. The change of ER or PR from positive to negative was associated with worse overall survival and post-recurrent survival (log-rank; P < 0.001, both). A subgroup analysis of HR-positive patients (i.e., positive ER, PR, or both) in primary tumor and HR-negative in metastatic sites showed that patients who accepted both salvage endocrine therapy and chemotherapy had better post-recurrent survival than did those who accepted salvage chemotherapy only (log-rank; P = 0.003). CONCLUSION: ER and PR status may change several times during metastatic tumor progression. A change of HR from positive to negative was associated with worse survival compared with consistent positivity. Repeated evaluations of HR status are necessary in metastatic breast cancer. Salvage hormonal therapy is still worth trying for patients whose HR status changes from positive to negative.

Cystic brain metastasis is associated with poor prognosis in patients with advanced breast cancer.

PURPOSE: Brain metastasis (BM) with a cystic component from breast cancer is rare and largely uncharacterized. The purpose of this study was to identify the characteristics of cystic BM in a large cohort of breast cancer patients. RESULTS: A total of 35 eligible patients with cystic BM and 255 patients with solid BM were analyzed. Three factors were significantly associated with an increased probability of developing cystic lesions: age at diagnosis ≤ 40 years, age at BM ≤ 45 years, and poor histological grade (p < 0.05). Patients with cystic metastasis were also characterized by a larger metastasis volume, a shorter progression-free survival (PFS) following their first treatment for BM, and poor overall survival after BM (p < 0.05). Multivariate analysis further demonstrated that local control of cystic BM was only potentially achieved for HER2-negative primary tumors (p = 0.084). METHODS: Breast cancer patients with parenchymal BM were reviewed from consecutive cases treated at our institution. Cystic BM was defined when the volume of a cystic lesion was greater than 50% of the aggregated volume of all lesions present. Clinicopathologic and radiographic variables were correlated with development of cystic lesions and with prognosis of cystic BM. CONCLUSIONS: This study shows that cystic BM from breast cancer, a special morphological type of BM, had worse prognosis than the more commonly observed solid BM. Younger age and low tumor grade were associated with the development of cystic lesions. Further comprehensive research and management of cystic BM are warranted to improve its poor prognosis.

Sunitinib as salvage treatment including potent anti-tumor activity in carcinomatous ulcers for patients with multidrug-resistant metastatic breast cancer.

OBJECTIVE: To evaluate the efficacy and safety of single-agent sunitinib as salvage treatment in Chinese patients with multidrug-resistant metastatic breast cancer (MBC). RESULTS: 37 patients were enrolled with median age of 48 years. 17 had hormone receptor (HR)-positive tumors, 7 had HER2-positive tumors, and 10 had triple-negative tumors. Among 32 evaluable patients with follow-up, 6 (18.8%) achieved partial response, 14 (43.8%) achieved stable disease, and 11 (34.4%) exhibited tumor shrinkage. The response rate in 9 patients with carcinomatous ulcers was 77.8%. The median progression free survival (PFS) was 8.6 weeks. Patients with a better response had improved overall survival and PFS relative to patients with a worse response (p = 0.007, p < 0.001). Compared with HR-negative tumor, HR-positive tumor had significantly better response to sunitinib (p = 0.035). The most frequent non-hematologic adverse events were fatigue (82.8%) and hypertension (34.5%). Grade 3/4 hematologic toxicity included neutropenia (82.8%) and thrombocytopenia (79.3%). There was no correlation between the clinical response and IHC findings. MATERIALS AND METHODS: Patients with MBC who were resistant to multiple salvage regimens (≥ 3 previous chemotherapy lines) were enrolled to receive sunitinib monotherapy. Dosage adjustment was allowed depending on adverse events. 14 patients underwent immunohistochemistry (IHC) testing for VEGF, PDGFR, EGFR and c-KIT. CONCLUSIONS: Sunitinib salvage treatment provided modest antitumor effect to patients with refractory multidrug-resistant MBC, especially to those with troublesome carcinomatous ulcers. The treatment-related adverse events of sunitinib were manageable through dosage adjustment.

Combined treatment with everolimus and fulvestrant reversed anti-HER2 resistance in a patient with refractory advanced breast cancer: a case report.

BACKGROUND: Everolimus, an inhibitor of the mammalian target of rapamycin, shows promising antitumor activity when combined with trastuzumab and chemotherapy for human epidermal growth factor receptor-2 (HER2)-positive breast cancer or when combined with endocrine agents for hormone receptor (HR)-positive tumors. However, data are limited regarding the effect of everolimus in combination with endocrine drugs in HER2-positive advanced breast cancer regardless of the HR status. CASE PRESENTATION: A 44-year-old female was diagnosed with recurrent HER2-positive breast cancer. The primary tumor was HR positive; however, the metastatic tumor was HR negative. The patient was resistant to classical chemotherapeutic agents and anti-HER2 treatment. Thus, the combination of everolimus and fulvestrant, a selective estrogen receptor downregulator, was chosen to reverse the resistance to anti-HER2 therapy. Indeed, the patient experienced long-term disease stabilization. Adverse events associated with the treatment were manageable by dose adjustments. We performed genetic testing of the metastatic tumor, which harbored a PIK3CA gene mutation but was positive for phosphatase and tensin homologue expression, which might result in resistance to the mammalian target of rapamycin inhibitor. CONCLUSION: This case study indicates that combined treatment with everolimus and fulvestrant might be a viable option for the treatment of metastatic breast cancer patients who are HER2 positive and carry a PIK3CA gene mutation but are resistant to anti-HER2 therapy and classical chemotherapeutic agents. Further prospective randomized trials are needed to confirm this finding.

Incidence and relapse risk of intracranial metastases within the perihippocampal region in 314 patients with breast cancer.

PURPOSE: The safe prerequisite of hippocampal-sparing whole brain radiotherapy (HS-WBRT) for patients with breast cancer is unclear. This study investigated the risk and relapse of perihippocampal (PH) metastases in breast cancer. METHODS: Consecutive breast cancer patients with brain metastasis (BM) were reviewed. Metastases and hippocampi were contoured in cranial magnetic resonance imaging (MRI). The closest distance from metastasis to hippocampus was calculated. Clinical and radiographic variables were correlated with PH (in or within 5mm around the hippocampus) metastasis. The risk of post-treatment PH recurrence was estimated. RESULTS: Three hundred and fourteen patients with 1678 metastases exhibited a median breast cancer-specific overall survival (OS) and OS after BM (BMOS) of 75.4 and 14.3 months, respectively. Hippocampal metastases were identified in 1.2% of metastases and 4.1% of patients. PH lesions comprised 3.5% of lesions in 11.1% of patients. The number and aggregated volume of BM were associated with PH disease probability (univariate). Only the number of BM significantly correlated with PH disease in the multivariate analysis. The patients with PH lesions exhibited more non-oligometastatic disease, increased tumor volume, and poor BMOS. One hundred and eleven patients without original PH lesions developed intracranial progression post-treatment. The risks of PH metastasis recurrence were 4.6% for WBRT and 6.8% for sub-therapeutic irradiation in the PH region. The increase in the absolute risk of PH recurrence with hippocampal-sparing irradiation was approximately 2%. CONCLUSIONS: These novel findings indicate that BM from breast cancer exhibits low risks of metastases and relapse within the hippocampal avoidance region. Non-oligometastatic disease is associated with PH metastasis. Thus, HS-WBRT is considered safe and suitable for breast cancer.

[PTEN loss correlates withthe clinical efficacy of lapatinib in HER2 positive metastatic breast cancer with trastuzumab-resistance].

OBJECTIVE: To discuss the indexes related to the efficacy of lapatinib after failure in trastuzumab in HER2-positive metastatic breast cancer (MBC) such as the status of PTEN, p-4EBP1 and clinical features. METHODS: Sixtymatched patients were included. Immunohistochemical (IHC) test of tissue specimens of metastatic lesions were applied to determine the status of PTEN and p-4EBP1. The correlation betweenclinical efficacy andthestatus of PTEN, p-4EBP1 and clinical features were analysed by long-rank test and Cox regression. RESULTS: In all patients, themedian progression free survival (PFS), ORR and CBR was 4.6 months, 36.7% and 50.0% respectively.Univariate analysis revealed that lapatinib-treated patients with PTEN loss (P = 0.015) and liver metastasis (P = 0.02) had significantly shorter median PFS. Multivariate analysis revealed that patients with PTEN lossandliver metastasis had higher risk of diseaseprogression (P = 0.005, 0.006, respectively). CONCLUSION: HER2-positive MBC with trastuzumab-resistance could benefit from lapatinib regimen. PTEN statusand liver metastasis couldpredict theclinical efficacyof subsequent lapatinib therapy. The detection ofrelated biomarkers could provide some referenceto optimize the personal treatment with HER2-positive breast cancer.

[Clinical efficacy and safety of trastuzumab plus capecitabine as first-line therapy for HER-2 positive metastatic breast cancer].

OBJECTIVE: To evaluate the efficacy and safety of trastuzumab plus capecitabine as first-line therapy for HER-2-positive metastatic breast cancer patients who have previously received anthracyclines and taxanes. METHODS: The patients who have HER-2-positive breast cancer recurrence and metastasis after treated by anthracyclines and taxanes were enrolled in this study. The patients received trastuzumab (6 mg/kg every 21 days following a loading dose of 8 mg/kg on cycle 1) and capecitabine (2 000 mg/m², days 1 to 14 every 21 days). RESULTS: 38 patients were enrolled. The median PFS for trastuzumab plus capecitabine was 8.6 months. The objective response rate (ORR) was 31.6%. Clinical benefit rate (CBR) was 65.8%. The most serious adverse event was leucopenia in one patient. Relatively common hematology toxicity was grade I or II leucopenia and neutropenia. The most common nonhematologic toxicity was hand-foot syndrome (HFS), which observed in 8 patients (21%). The morbidity of grade II and III HFS were 8% and 3% respectively. CONCLUSION: Trastuzumab plus capecitabine is an effective and safe regimen as first-line treatment for HER-2-positive metastatic breast cancer patients whose cancer is resistant to treatment with anthracyclines and taxanes.

National consensus in China on diagnosis and treatment of patients with advanced breast cancer.

The recently available guidelines on the management of advanced breast cancer (ABC) organized by Chinese Anti-Cancer Association, Committee of Breast Cancer Society (CACA-CBCS) do not elucidate ABC in details. To instruct clinicians in treatment of ABC, a Chinese expert consensus meeting on diagnosis and treatment of ABC was held in June 2014 and a consensus is developed. The following consensus provides the level of evidence and supporting documents for each recommendation, and introduces research topics to be urgently addressed. Notably, the consensus on diagnosis and treatment of ABC in China is developed to be applied nationwide. In different areas, multidisciplinary treatment (MDT) tailored to the each patient and the disease itself should be applied based on the basic principles of modern oncology.

Classification of Epidermal Growth Factor Receptor Gene Mutation Status Using Serum Proteomic Profiling Predicts Tumor Response in Patients with Stage IIIB or IV Non-Small-Cell Lung Cancer.

OBJECTIVES: Epidermal growth factor receptor (EGFR) gene mutations in tumors predict tumor response to EGFR tyrosine kinase inhibitors (EGFR-TKIs) in non-small-cell lung cancer (NSCLC). However, obtaining tumor tissue for mutation analysis is challenging. Here, we aimed to detect serum peptides/proteins associated with EGFR gene mutation status, and test whether a classification algorithm based on serum proteomic profiling could be developed to analyze EGFR gene mutation status to aid therapeutic decision-making. PATIENTS AND METHODS: Serum collected from 223 stage IIIB or IV NSCLC patients with known EGFR gene mutation status in their tumors prior to therapy was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and ClinProTools software. Differences in serum peptides/proteins between patients with EGFR gene TKI-sensitive mutations and wild-type EGFR genes were detected in a training group of 100 patients; based on this analysis, a serum proteomic classification algorithm was developed to classify EGFR gene mutation status and tested in an independent validation group of 123 patients. The correlation between EGFR gene mutation status, as identified with the serum proteomic classifier and response to EGFR-TKIs was analyzed. RESULTS: Nine peptide/protein peaks were significantly different between NSCLC patients with EGFR gene TKI-sensitive mutations and wild-type EGFR genes in the training group. A genetic algorithm model consisting of five peptides/proteins (m/z 4092.4, 4585.05, 1365.1, 4643.49 and 4438.43) was developed from the training group to separate patients with EGFR gene TKI-sensitive mutations and wild-type EGFR genes. The classifier exhibited a sensitivity of 84.6% and a specificity of 77.5% in the validation group. In the 81 patients from the validation group treated with EGFR-TKIs, 28 (59.6%) of 47 patients whose matched samples were labeled as "mutant" by the classifier and 3 (8.8%) of 34 patients whose matched samples were labeled as "wild" achieved an objective response (p<0.0001). Patients whose matched samples were labeled as "mutant" by the classifier had a significantly longer progression-free survival (PFS) than patients whose matched samples were labeled as "wild" (p=0.001). CONCLUSION: Peptides/proteins related to EGFR gene mutation status were found in the serum. Classification of EGFR gene mutation status using the serum proteomic classifier established in the present study in patients with stage IIIB or IV NSCLC is feasible and may predict tumor response to EGFR-TKIs.

HPIP is upregulated in colorectal cancer and regulates colorectal cancer cell proliferation, apoptosis and invasion.

Hematopoietic pre-B cell leukemia transcription factor (PBX)-interacting protein (HPIP) was shown to play a role in cancer development and progression. However, the role of HPIP in colorectal cancer (CRC) is unknown. Here, we report that HPIP is overexpressed in most of CRC patients and predicts poor clinical outcome in CRC. HPIP promotes CRC cell proliferation via activation of G1/S and G2/M checkpoint transitions, concomitant with a marked increase of the positive cell cycle regulators, including cyclin D1, cyclin A, and cyclin B1. HPIP inhibits CRC cell apoptosis accompanied by the decreased levels of BAX and PIG3, the inducers of apoptosis, and the increased level of the apoptosis inhibitor BCL2. HPIP blocks caspase-3-mediated cleavage of PARP, an important apoptosis marker. HPIP promotes CRC cell migration and invasion, and regulates epithelial-mesenchymal transition (EMT), which plays a critical role in cancer cell migration and invasion. Activation of MAPK/ERK1/2 and PI3k/AKT pathways is required for HPIP modulation of CRC cell proliferation, migration and EMT. Moreover, HPIP knockdown suppresses colorectal tumor growth in nude mice. These data highlight the important role of HPIP in CRC cell proliferation and progression and suggest that HPIP may be a useful target for CRC therapy.

[Detection of circulating tumor cells and its clinical value for different stages and various subtypes of breast cancer].

OBJECTIVE: To explore the prognostic value of applying a semi-automated detection system for detecting circulating tumor cells (CTC) for different stages and various subtypes of breast cancer. METHODS: Immunomagnetic separation and immunofluorescent staining were employed to detect the expressions of CTC and CTC HER-2. Chi-square test, univariate and multivariate analyses were used to examine the correlations between CTC and clinical characteristics. RESULTS: CTC was detected in 57.5% (138/240) of metastatic breast cancer (MBC). CTC enumeration, HER-2 status, number of metastasis and bone metastasis were relevant (P < 0.05). CTC was detected in 23.1% (6/26) of early-stage breast cancer (EBC). CTC enumeration, pathological type, estrogen receptor (ER), progesterone receptor (PR), Ki-67, tumor size and lymph node metastasis were irrelevant (P > 0.05). HER-2-positive MBC detected less CTC than HER-2-negative (χ(2) = 12.296, P < 0.001) . CTC HER-2 expression was different between HER-2-positive and negative patients in 99 MBC cases (χ(2) = 8.082, P = 0.004). CONCLUSION: CTC enumeration is different for various stages and different subtypes of breast cancer. CTC enumeration and bone metastasis are relevant. CTC detection in EBC may predict tumor relapse. And CTC HER-2 expression is different between HER-2-positive and negative MBC patients.

Schlafen-11 sensitizes colorectal carcinoma cells to irinotecan.

Schlafen-11 (SLFN11) showed a highly significant positive correlation with the response of topoisomerase inhibitors in cancer cell lines derived from prostate, lung, etc. However, this finding has not been validated in colorectal cancers (CRCs). Although irinotecan (CPT-11), a topoisomerase inhibitor, is one of the most important drugs in the treatment of advanced and/or metastatic CRC, resistance is a critical drawback to its clinical effectiveness. The present study aimed to investigate the mechanism of SLFN11 in the response of CRC cell lines to SN-38 (an active CPT-11 metabolite) treatment. Western blotting was used to measure protein expression levels of SLFN11 in human CRC cell lines. Then, SLFN11 expression was modulated by transfecting human CRC cell lines with vectors carrying the SLFN11 gene or specific SLFN11 small interfering RNAs. The effects of SN-38 treatment on CRC cells with different SLFN11 expression levels were detected, including inhibition of cell growth, induction of apoptosis, and cell cycle arrest. This study showed that SLFN11 expression varied between the CRC cell lines and high-level SLFN11 expression promoted SN-38-induced antiproliferative activity, apoptosis, and cell cycle arrest. Our results suggest that SLFN11 plays a key role in cell cycle arrest and/or induction of apoptosis in response to exogenous SN-38-induced DNA damage and might be used as a new predictive biomarker for CRC treatment.

[Serum HER2 ECD level and its clinical significance in advanced breast cancer patients with different molecular subtypes].

OBJECTIVE: To evaluate the serum HER2 ECD level and its significance in advanced breast cancer patients with different molecular subtypes. METHODS: A total of 322 advanced breast cancer patients were enrolled. The serum HER2 ECD concentrations were quantitatively detected by enzyme-linked immunosorbent assay(ELISA). Its relationship with clinicopathological characteristics and clinical significance were analyzed. RESULTS: It was found that 55.9% (19/34)Luminal A, 42.7% (44/103) Luminal B-HER2(-), 70.6% (60/85) Luminal B-HER2(+), 73.8% (45/61)HER2-enriched and 23.1% (9/39) triple-negative patients had serum concentrations of HER2 ECD at least 15 ng/ml respectively. The prevalence of elevated ECD level in patients of different molecular subtypes differed significantly (P < 0.001). Tissue HER2 status, number of metastatic sites, visceral metastasis, CA15-3 and carcinoembryonic antigen(CEA) levels exhibited statistically significant correlations with the prevalence of elevated serum HER2 ECD level. The serum concentrations of HER2 ECD decreased after effective targeted therapy in tissue HER2-positive patients. CONCLUSION: The prevalence of elevated serum levels of HER2 ECD differed significantly in advanced breast cancer patients with different molecular subtypes. The serum HER2 ECD level may reflect both histological HER2 status and tumor load. And the dynamic changes of ECD concentrations are somewhat correlated with the efficacies of targeted treatment.

[Clinical observations of ductal carcinoma in situ and early stage breast cancer with invasive depth less than 1 cm].

OBJECTIVE: To explore the clinical characteristics and prognosis of ductal carcinoma in situ (DCIS) and early stage ductal breast cancer with invasive depth ≤ 1 cm. METHODS: Follow-up analyses were performed for the clinical data of 57 patients with DCIS, 46 with pT(1mic) and 74 with pT(1a-b) breast cancer treated or consulted at our hospital. Single factor analysis was used to examine their prognostic factors. RESULTS: Among them, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER-2) positive rate and visual tumor size (2.0(0.1-7.0) vs 2.0(0.5-10.0) vs 2.0(0.3-10.0) cm)had no statistical differences between 3 groups (all P > 0.05). After median follow-up periods of 63, 38, 59 months, 12 patients suffered recurrence and metastasis and the rate of each group had no statistical differences. For pT(1a-b) patients with positive hormone receptor, endocrine therapy markedly reduced the risk of recurrence and metastasis (P = 0.024) . pT(1mic), pT(1a-b) patients on chemotherapy had higher or comparable recurrence rate versus those without. And DCIS patients on chemotherapy had a much higher recurrence rate (P = 0.016) . In pT(1a-b) group, HER-2 positive patients had higher recurrence and metastasis rates. Yet there was a decreasing tendency after Herceptin treatment. CONCLUSIONS: Chemotherapy without proper indications may not improve the prognosis of DCIS, pT(1mic) and pT(1a-b) patients. Endocrine therapy is the crucial prognostic factor of patients with positive hormone receptor. Use of Herceptin for HER-2-positive patients is probably significant.

Ki-67 index as a prognostic factor of subsequent lapatinib-based therapy in HER2-positive metastatic breast cancer with resistance to trastuzumab.

Prognostic factor analysis has been conducted to determine whether the parameters of clinical data and biomarkers would predict differential progression-free survival (PFS) or overall survival (OS) from lapatinib-based therapy in patients with primary or acquired resistance to trastuzumab. Treatment with lapatinib plus capecitabine for HER2-positive metastatic breast cancer (MBC) with primary or acquired resistance to trastuzumab was analyzed retrospectively. Tumor biomarkers, which came from the biopsies before the starting of lapatinib therapy, were evaluated by immunohistochemistry (IHC). Prognostic factors related to PFS or OS of the lapatinib therapy were assessed by univariate and multivariate analysis. Ki-67 index and liver metastases were the significant prognostic factors for predicting PFS of subsequent lapatinib therapy in the univariate analysis and the multivariate analysis. The risk for disease progression in patients who had a Ki-67 index<40% was 59% less than that in patients had Ki-67 ≥ 40 (HR = 0.41, 95% CI, 0.23-0.74, P = 0.003). TTP of prior trastuzumab therapy, liver metastases, and the number of metastatic sites were three independent prognostic factors of subsequent lapatinib therapy. Ki-67 index was the significant prognostic factors for predicting PFS of the subsequent second line targeted therapy in patients with trastuzumab resistance.

PTEN mutation, methylation and expression in breast cancer patients.

The tumor suppressor gene, PTEN, has previously been demonstrated to be involved in breast tumorigenesis and tumor progression. The aim of the present study was to investigate the expression and significance of PTEN in breast carcinomas, to detect the mutation frequency of PTEN in sporadic breast carcinoma tissues and to determine the association between PTEN promoter methylation and gene expression. Immunohistochemical methods were used to analyze the expression of the PTEN gene in 146 cases of breast carcinoma and 10 cases of normal breast tissue closely adjacent to the carcinoma. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis was used to analyze conformation polymorphisms in 45 breast carcinoma and 10 normal breast tissues. Point mutations of abnormal single stranded conformation were detected by DNA sequencing. The methylation of the PTEN promoter was analyzed by methylation-specific PCR. Expression of PTEN was detected in 57.5% (84/146) of patients with breast carcinoma. By contrast, PTEN expression was detected in 100% of normal samples. Expression of PTEN was found to negatively correlate with the tumor size, the pathological stage and the expression of the estrogen receptor (ER) and the progesterone receptor (PR) in breast cancer. The 2-year disease-free survival of patients with a high expression of PTEN was higher compared with those with low PTEN expression (P<0.05). Missense mutations in exon 2 of PTEN were identified in 1/45 breast cancer cases. PTEN promoter methylation was detected in 31.1% (14/45) of breast carcinomas, of which 64.3% (9/14) were associated with a loss of PTEN expression. The tumor suppressor gene, PTEN, was abnormally expressed in the breast carcinomas. The number of PTEN mutations were low (1/45) in the sporadic breast cancer cases analyzed in the present study and PTEN promoter methylation may have been the main mechanism leading to the decreased expression of PTEN. These results indicate that PTEN is important for the tumorigenesis, development and prognosis of breast cancer.