[Value of intraoperative GeneSearch(TM) BLN assay to detect breast cancer metastases in sentinel lymph nodes].

OBJECTIVE: To evaluate the value of GeneSearch(TM) BLN assay as an intraoperative diagnostic method of sentinel lymph node metastases in breast cancer patients. METHODS: Ninety consecutive patients were involved in this study. SLNs were intraoperatively identified and dissected, and then sectioned vertically to the long axis into multiple blocks. The odd blocks were tested by BLN assay and even ones prepared for frozen sectioning (FS), while all blocks were evaluated by touch imprint cytology (TIC). Post-operatively, residual tissues of the even blocks were assessed by histopathologic examination (4 - 6 µm thick serial sectioning permanent H&E slides were performed every 150 µm and one block made 6 slides). RESULTS: BLN assay could be performed within less than 35 min after learning curve of 10 cases. A correlation was found between cycle time values of mammaglobin or cytokeratin-19 and size of metastases, with Spearman correlation coefficients of 0.67 and 0.71, respectively. The accuracy, sensitivity, specificity, positive predict value (PPV) and negative predict value (NPV) of the assay were 95.6%, 93.3%, 96.7%, 93.3% and 96.7%, While FS had the sensitivity, specificity, PPV, NPV of 76.7%, 100%, 100%, 89.6%, and TIC of 73.3%, 100%, 100%, 88.2%, respectively. The sensitivity of the assay was higher than that of FS (P = 0.07), and was significantly higher than that of FS (P = 0.04). When assessing patients with micro-metastases, the assay had a sensitivity of 85.7%, which was significantly higher than that of FS and TIC (P = 0.03). CONCLUSION: GeneSearch(TM) BLN Assay can replace FS and TIC for the intraoperative assessment of SLN.

Distributions of angiogenesis and lymphangiogenesis in gastrointestinal intramucosal tumors.

BACKGROUND: Although angiogenesis and lymphangiogenesis in gastrointestinal cancers has been investigated in many studies, their distribution characteristics in gastrointestinal intramucosal tumors have not been well addressed. METHODS: We evaluated the blood microvessel density (BMVD) and lymphatic microvessel density (LMVD) by immunostaining with monoclonal antibodies of CD34 and D2-40 in 37 patients with stomach intramucosal carcinoma and 28 patients with colorectal intramucosal neoplasia. Microvessels with endothelial cells labeled by CD34 but not by D2-40 were recognized as blood microvessels; and microvessels with endothelial cells labeled by both CD34 and D2-40 were recognized as lymphatic vessels. Furthermore, the relationships between expression of vascular endothelial growth factor (VEGF), VEGF-C, and BMVD, LMVD were investigated as well. RESULTS: The LMVD was significantly higher in peritumoral tissues than in corresponding normal tissues in gastrointestinal intramucosal tumors (20.87 versus 14.56, P = 0.003). However, there was no significant difference in the BMVD between peritumoral tissues and corresponding normal tissues (P = 0.166). The BMVD in peritumoral tissues was higher in patients with lymph node metastases than in patients without lymph nodes metastases (P = 0.047). Our results did not show significant association between VEGF, VEGF-C and BMVD, LMVD. CONCLUSIONS: Our results suggested that the increase of lymphangiogenesis seems superior to the increase of angiogenesis in gastrointestinal intramucosal tumors.

[Correlation of VEGF and Ki67 expression with sensitivity to neoadjuvant chemoradiation in rectal adenocarcinoma].

OBJECTIVE: To investigate the correlation of expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (Ki67) with sensitivity to neoadjuvant chemoradiation in rectal adenocarcinoma. METHODS: Samples of pretreatment biopsies and the resected specimens after neoadjuvant therapy in 32 patients with rectal adenocarcinoma were collected, and the expression of Ki67 and VEGF were detected by immunohistochemistry using specific antibodies. The correlation of Ki67 and VEGF expression with clinicopathological factors were analyzed. RESULTS: The level of VEGF expression was significantly correlated with lymph node metastasis (P = 0.033), depth of tumor invasion (P = 0.007) and TNM stage (P = 0.016), but not with histological type, tumor size, age and gender of the patients (P > 0.05). However, VEGF expression was found to be negatively and significantly correlated with the sensitivity to neoadjuvant chemoradiation (P = 0.016), and a transient increase of VEGF expression was detected in the resected specimens after neoadjuvant therapy (P = 0.035). Ki67 labeling index (Ki67-LI) was found to be significantly correlated with lymph node metastasis (P = 0.028), but not with tumor size, age and gender of the patients (P > 0.05). It was also found that tumors with lower Ki67-LI expression were more sensitive to neoadjuvant therapy than that with higher expression of Ki67-LI (P = 0.032). In contrast with VEGF, the Ki67 expression level decreased after neoadjuvant therapy, but no statistical significance was found between pretreatment and posttreatment specimens (P > 0.05). CONCLUSION: The preliminary results of this study demonstrate that the expression of VEGF and Ki67 in pretreatment biopsy of rectal adenocarcinoma may be used as a biomarker to predict tumor response to neoadjuvant chemoradiation.

[Molecular imaging of epidermal growth factor receptor in glioma-bearing rats].

OBJECTIVE: To investigate the value of 11C-PD153035 as an EGFR imaging agent in C6 tumor-bearing rat. METHODS: The tumor-bearing rats were generated by subcutaneous injection of glioma C6 cells. Positron emission tomography/computer tomography (PET/CT) scans started as soon as intravenous injection of 11C-PD153035 (15-20 MBq/0.3 ml) was completed, images were collected continuously. The region of interest (ROI) was used to study the percentage of radioactivity in major organs and implanted tumors in the rats. The accumulation and blocking study in vitro was completed. RESULTS: There were significant differences in 11C-PD153035 uptake among major organs. The maximum uptake in the organs ranked in the following order: liver > gastrointestinal tract > kidney > lung > brain > muscle. Radioactivity could be also observed in the tumors. The radioactivity ratio (T/NT, target/non-target) peaked (4.15) at 40 - 50 min post injection. The in vitro blocking study showed that 11C-PD153035 uptaken by C6 cells could be blocked by PD153035. CONCLUSION: The results of this study show that 11C-PD153035 can be uptaken by EGFR-expressing tumors. 11C-PD153035 has a potential as a bioprobe to yield useful information for both diagnosis and therapy of tumors. However, the high concentration of 11C-PD153035 in the gastrointestinal tract is unfavorably affecting the tumor detection in these organs.

[Correlation between FDG PET /CT and the expression of glutl and ki-67 antigen in esophageal cancer].

OBJECTIVE: To evaluate the correlation between standardized uptake valus (SUV) of 18F-fluorodeoxyglucose (18 F-FDG) of tumor at PET/CT examination and the expression of glucose transporter-1 (Glutl) and Ki-67 in esophageal cancer. METHODS: 56 patients with esophageal cancer were evaluated with 18 F-FDG PET/CT examination before operation. The expression of Glut1 and Ki-67 antigen in the tumor tissues was detected by immunohistochemistry after operation. RESULTS: (1) Positive rate of Glutl and Ki-67 expression in esophageal cancer tissues was 100% , respectively. There was a positive correlation between the expression of Glutl and Ki-67 and the clinical stages and differentiation of the tumor. The more the tumor and the clinical stages were advanced and the lower was the tumor differentiation, the more Glutl and Ki-67 were expressed. (2) There were abnormal radioactive high uptake regions on PET/CT imaging of esophagus in the 56 patients, which were confirmed by pathology as the primary carcinoma. The SUV was higher than 2. 5. There was a gradually increasing tendency in SUV along with the lowering of the tumor differentiation and the advance of clinical stages. (3)There was a correlation between the expression of Glutl, Ki-67 and the SUV, the more Glutl and Ki-67 were expressed, the higher the SUV of tumor 18F-FDG at PET/CT examination was in esophageal tumor tissues. CONCLUSION: There is a widespread expression of Glutl in esophageal cancer tissues, and the SUV may be used to indirectly evaluate the proliferative capacity of esophageal cancer.

Proliferation PET image to characterize pathological spatial features in patients with non-small cell lung cancer: a pilot study.

PURPOSE: (18)F-FLT-PET imaging was proposed as a tool for measuring in vivo tumor cell proliferation and detecting sub-volumes to propose escalation in radiotherapy. The aim of this study was to validate whether high FLT uptake areas in (18)F-FLT PET/CT are coincident with tumor cell proliferation distribution indicated by Ki-67 staining in non-small cell lung cancer, thus provide theoretical support for the application of dose painting guided by (18)F-FLT PET/CT. MATERIALS AND METHODS: Twelve treatment naive patients with biopsy proven NSCLC underwent (18)F-FLT PET/CT scans followed by lobectomy were enrolled. The surgical specimen was dissected into 4-7 µm sections at approximately 4-mm intervals. The best slice was sort out to complete Ki-67 staining. Maximum Ki-67 labelling Index and SUVmax of the corresponding PET image was calculated. The correlation between Ki-67 Labelling Index and SUVmax of FLT was determined using Spearman Correlation analysis. High uptake areas and high proliferating areas were delineated on the two images, respectively, and their location was compared. RESULTS: The maximal SUV was 3.26 ± 0.97 (1.96-5.05), maximal Ki-67 labeling index was 49% ± 27.56% (5%-90%). Statistical analysis didn't reveal a significant correlation between them (r = -0.157, P = 0.627, > 0.05). 9 patients can contour high proliferating area on Ki-67 staining slice, and eight can contour the high uptake areas. In 4 patients, we can observe a generally close distribution of high uptake areas and high proliferating areas, in one patient, both the uptake level and proliferation status was low, while the others didn't not find a significant co-localization. CONCLUSION: Noninvasive (18)F-FLT PET assessing the proliferative status may be a valuable aid to guide dose painting in NSCLC, but it needs to be confirmed further.

PDCD4 as a predictor of sensitivity to neoadjuvant chemoradiotherapy in locally advanced rectal cancer patients.

OBJECTIVE: The purpose of this study was to examine the role of programmed cell death 4 (PDCD4) expression in predicting tumor response to neoadjuvant chemoradiotherapy and outcomes for patients with locally advanced rectal cancer. METHODS: Clinicopathological factors and expression of PDCD4 were evaluated in 92 patients with LARC treated with nCRT. After the completion of therapy, 4 cases achieved clinical complete response (cCR), and thus the remaining 88 patients underwent a standardized total mesorectal excision procedure. There were 38 patients (41.3%) with a good response (TRG 3-4) and 54 (58.7%) with a poor one (TRG 0-2). RESULTS: Immunohistochemical staining analyses showed that patients with high expression of PDCD4 were more sensitive to nCRT than those with low PDCD4 expression (P=0.02). High PDCD4 expression before nCRT and good response (TRG3-4) were significantly associated with improved 5-year disease-free survival and 5-year overall survival (P<0.05). Multivariate analysis demonstrated that the pretreatment PDCD4 expression was an independent prognostic factor. CONCLUSION: Our study demonstrated that high expression of PDCD4 protein is a useful predictive factor for good tumor response to nCRT and good outcomes in patients with LARC.

Circulating lymphocytes as predictors of sensitivity to preoperative chemoradiotherapy in rectal cancer cases.

OBJECTIVE: The objective of this study was to identify clinical predictive factors for tumor response after neoadjuvant chemoradiotherapy (nCRT) in locally advanced rectal cancer (LARC). METHODS: All factors were evaluated in 88 patients with LARC treated with nCRT. After a long period of 4-8 weeks of chemoradiotherapy, 3 patients achieved clinical complete response (cCR) and thus aggressive surgery was avoided, and the remaining 85 patients underwent a curative-intent operation. The response to nCRT was evaluated by tumor regression grade (TRG) system. RESULTS: There were 32 patients (36.4%) with good tumor regression (TRG 3-4) and 56 (63.6%) with poor tumor regression (TRG 0-2). Lymphocyte counts and ratios were higher in good response cases (P=0.01, 0.03, respectively) while neutrophil ratios and N/L ratios were higher in poor response cases (P=0.04, 0.02, respectively). High lymphocyte ratios before nCRT and good tumor regression (TRG3-4) were significantly associated with improved 5-year disease-free survival (P<0.05). Pretreatment nodal status was also significantly associated with 5-year disease-free survival and 5-year overall survival (P<0.05). Multivariate analysis confirmed that the pretreatment lymphocyte ratio and lymph nodal status were independent prognostic factors. CONCLUSION: Our study suggested that LARC patients with high lymphocyte ratios before nCRT would have good tumor response and high 5-year DFS and OS.

A comparative proteomic study identified calreticulin and prohibitin up-regulated in adrenocortical carcinomas.

BACKGROUND: Identifying novel tumor biomarkers to develop more effective diagnostic and therapeutic strategies for patients with ACC is urgently needed. The aim of the study was to compare the proteomic profiles between adrenocortical carcinomas (ACC) and normal adrenocortical tissues in order to identify novel potential biomarkers for ACC. METHODS: The protein samples from 12 ACC tissues and their paired adjacent normal adrenocortical tissues were profiled with two-dimensional electrophoresis; and differentially expressed proteins were identified by mass spectrometry. Expression patterns of three differently expressed proteins calreticulin, prohibitin and HSP60 in ACC, adrenocortical adenomas (ACA) and normal adrenocortical tissues were further validated by immunohistochemistry. RESULTS: In our proteomic study, we identified 20 up-regulated and 9 down-regulated proteins in ACC tissues compared with paired normal controls. Most of the up-regulated proteins were focused in protein binding and oxidoreductase activity in Gene Ontology (GO) molecular function classification. By immunohistochemistry, two biomarkers calreticulin and prohibitin were validated to be overexpressed in ACC compared with adrenocortical adenomas (ACA) and normal tissues, but also calreticulin overexpression was significantly associated with tumor stages of ACC. CONCLUSION: For the first time, calreticulin and prohibitin were identified to be novel candidate biomarkers for ACC, and their roles during ACC carcinogenesis and clinical significance deserves further investigation. VIRTUAL SLIDES: The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1897372598927465.

Using intra-operative GeneSearchTM Breast Lymph Node Assay to detect breast cancer metastases in sentinel lymph nodes: results from a single institute in China.

BACKGROUND: Sentinel lymph node (SLN) biopsy has become a common procedure for early breast cancer patients. The GeneSearch(TM) Breast Lymph Node (BLN) Assay is a real-time RT-PCR assay for the detecting nodal metastases larger than 0.2 mm. China Breast Cancer Clinical Study Group (CBCSG)-001a is a prospective multi-center clinical trial that was conducted to validate the GeneSearch(TM) BLN Assay in China. METHODS: The SLNs from 90 consecutive patients were identified and dissected, and then sectioned along the short axis into multiple blocks. Intra-operatively, the odd blocks were tested by BLN assay and the even ones were used for frozen section, while all the blocks were evaluated by touch imprint cytology. Post-operatively, the remaining tissues were assessed by histological evaluation. RESULTS: A total of 189 SLNs was tested by BLN assay. The sensitivity, specificity, positive predictive value, and negative predictive value were 88.9%, 97.4%, 88.9% and 97.4%, respectively, for BLN assay, 75.0%, 100%, 100% and 94.4%, respectively, for frozen section, and 63.9%, 100%, 100% and 92.2%, respectively, for touch imprint cytology. The sensitivity of BLN assay was higher than that of touch imprint cytology (P = 0.01) and frozen section (P = 0.13). When assessing the nodes with micro-metastases, BLN assay had a significant higher sensitivity than frozen section (P = 0.023) and touch imprint cytology (P = 0.005). CONCLUSION: The GeneSearch(TM) BLN Assay is an accurate and rapid intra-operative assay for breast SLNs and it is suitable for application in general medical practice.

[Comparison of the characteristics in recurrence and metastasis between bronchioloalveolar carcinoma and other lung adenocarcinomas].

BACKGROUND & OBJECTIVE: The histological definition of bronchioloalveolar carcinoma (BAC) has been changed recently by the revised World Health Organization (WHO) classification. Although bronchioloalveolar carcinoma is a subtype of lung adenocarcinoma, its biological features are better than those of other lung adenocarcinomas. This study was to analyze differences in metastatic activity between bronchioloalveolar carcinoma and other lung adenocarcinomas. METHODS: The expression of E-Cadherin, Collagen IV, vascular endothelial growth factor receptor-2 (VEGFR-2), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in 28 specimens of stage I bronchioloalveolar carcinoma confirmed pathologically and 40 specimens of other stage I lung adenocarcinomas were detected by immunohistochemistry. Their correlations to tumor recurrence and metastasis were analyzed. RESULTS: The 5-year survival rate was significantly higher in ths patients with bronchioloalveolar carcinoma than in the patients with other lung adenocarcinomas (88.7% vs. 57.3%, P < 0.05). The intrathoracic recurrence rate was significantly higher and the extrathoracic metastasis rate was significantly lower in the patients with bronchioloalveolar carcinoma than in the patients with other lung adenocarcinomas (75% vs. 33.3%, 25% vs. 66.7%, P < 0.05). The positive rates of Collagen IV, E-Cadherin and TIMP-1 were significantly higher in bronchioloalveolar carcinoma than in other lung adenocarcinomas (78.6% vs. 42.5%, 78.6% vs. 40.0%, 67.5% vs. 42.9%, all P < 0.01). The positive rate of VEGFR-2 was significantly higher in other lung adenocarcinomas than in bronchioloalveolar carcinoma (85.7% vs. 77.5%, P < 0.05). There was no significant difference in the positive rate of MMP-9 between bronchioloalveolar carcinoma and other lung adenocarcinomas (85.0% vs. 78.6%, P = 0.494). CONCLUSION: As compared with other lung adenocarcinomas, stage I bronchioloalveolar carcinoma is less aggressive in clinical behavior and likely to develop intrathoracic recurrence, with less extrathoracic metastases and better prognosis.