Chemotherapy-Induced IL8 Upregulates MDR1/ABCB1 in Tumor Blood Vessels and Results in Unfavorable Outcome.

Tumor endothelial cells (TEC) lining tumor blood vessels actively contribute to tumor progression and metastasis. In addition to tumor cells, TEC may develop drug resistance during cancer treatment, allowing the tumor cells to survive chemotherapy and metastasize. We previously reported that TECs resist paclitaxel treatment via upregulation of ABCB1. However, whether TEC phenotypes are altered by anticancer drugs remains to be clarified. Here, we show that ABCB1 expression increases after chemotherapy in urothelial carcinoma cases. The ratio of ABCB1-positive TEC before and after first-line chemotherapy in urothelial carcinoma tissues (n = 66) was analyzed by ABCB1 and CD31 immunostaining. In 42 cases (64%), this ratio increased after first-line chemotherapy. Chemotherapy elevated ABCB1 expression in endothelial cells by increasing tumor IL8 secretion. In clinical cases, ABCB1 expression in TEC correlated with IL8 expression in tumor cells after first-line chemotherapy, leading to poor prognosis. In vivo, the ABCB1 inhibitor combined with paclitaxel reduced tumor growth and metastasis compared with paclitaxel alone. Chemotherapy is suggested to cause inflammatory changes in tumors, inducing ABCB1 expression in TEC and conferring drug resistance. Overall, these findings indicate that TEC can survive during chemotherapy and provide a gateway for cancer metastasis. Targeting ABCB1 in TEC represents a novel strategy to overcome cancer drug resistance. SIGNIFICANCE: These findings show that inhibition of ABCB1 in tumor endothelial cells may improve clinical outcome, where ABCB1 expression contributes to drug resistance and metastasis following first-line chemotherapy.

A small-scale experimental study of breast FNA consultation on the internet using Panoptiq.

INTRODUCTION: To test the potential for cytopathology consultation using Panoptiq (ViewsIQ, Richmond, BC, Canada; this is a new type of whole-slide image that is made manually and incorporates video content), we investigated its application in the cytopathological diagnosis of cases that were difficult to diagnose by breast fine-needle aspiration (FNA). MATERIALS AND METHODS: Panoptiq files were created from liquid-based cytology slides prepared by the BD CytoRich Red (BD, Franklin Lakes, NJ) method. The slides were prepared from 23 consecutive samples of breast FNA that had been diagnosed as atypical or suspicious by the Hokkaido Cancer Center, Hokkaido, Japan. Nine volunteer reviewers, who were provided with the URL of the Panoptiq file, the original cytopathological diagnosis, and the clinical information, were asked to classify the cytopathological diagnosis of each case into 4 diagnostic categories (benign, atypical, suspicious, or malignant). We examined the consultation benefit (CB)-how much closer the reviewer's cytopathology diagnosis came to the final histopathological diagnosis than the original cytodiagnosis. The CB scoring system was decided in advance. RESULTS: All 9 reviewers showed a positive total CB score and 2 reviewers showed a significantly higher CB score (Wilcoxon's signed rank test). The representative diagnosis (ie, the most frequently rendered diagnosis in each case) also showed a significant CB. CONCLUSIONS: Our small-scale experimental study, in which Panoptiq was used in the diagnosis of cases that were difficult to diagnose definitively by breast FNA, revealed a positive CB score by every reviewer and the representative diagnosis showed a significant CB. The study suggests that Panoptiq could be used for cytopathology consultation.

Clinicopathological significance of PSF3 expression in uterine endometrial carcinomas.

PSF3 (Partner of SLD Five 3) is a member of the heterotetrameric complex termed GINS. Previous studies have shown that PSF3 is up-regulated in several cancers and is associated with tumor malignancy. However, the clinicopathological significance of PSF3 expression in endometrial lesions is still poorly understood. To investigate whether PSF3 could serve as a useful biomarker for endometrial carcinomas, we performed immunohistochemical analysis of PSF3 expression. In 155 cases of endometrial carcinomas (ECs), the mean tumor proportion score of PSF3 expression was 30.7% in G1 endometrioid carcinoma, 55.0% in G2 endometrioid carcinoma, 59.0% in G3 endometrioid carcinoma, and 58.9% in nonendometrioid carcinomas. In 25 cases of atypical hyperplasia, the mean tumor proportion score of PSF3 expression was significantly lower (10.4%). High expression of PSF3 was associated with more advanced pathologic T stage (P = .000), lymphatic invasion (P = .001), and poor clinical outcomes such as shorter relapse-free survival (P = .000) and overall survival (P = .001). When we compared the immunostaining of PSF3 and Ki-67, the proportions of PSF3-positive cells in tumor epithelial cells were comparable to those of Ki-67-positive cells. However, PSF3-positive cells were selectively found in tumor cells, whereas Ki-67-positive cells were also found in tumor stromal cells. These results demonstrated that PSF3 immunostaining was valuable as a histopathologic marker for differential diagnosis between atypical hyperplasia and ECs, for tumor histologic grading, and for determining a patient's prognosis. PSF3 may play a crucial role in tumor progression in EC.

The therapeutic candidate for immune checkpoint inhibitors elucidated by the status of tumor-infiltrating lymphocytes (TILs) and programmed death ligand 1 (PD-L1) expression in triple negative breast cancer (TNBC).

BACKGROUND: The status of tumor-infiltrating lymphocytes (TILs) is a prognostic factor for triple negative breast cancer (TNBC). Recent studies have shown that programmed cell death 1 (PD-1) or programmed death ligand 1 (PD-L1) is expressed on T lymphocytes or tumor cells modulating antitumor immunity. The regulation of immune checkpoints between tumor cells and T lymphocytes may serve as a target for improvement of TNBC prognosis. We investigated TILs and PD-L1 status in TNBCs before or after preoperative systemic therapy (PST) to elucidate the clinical significance of PD-L1 expression. METHODS: Ninety patients received PST, and materials of core needle biopsies (CNB) taken before PST were available for 32 patients. TILs were scored as "% stromal", and tumors were defined as High-TILs (≥30%) or Low-TILs (<30%). The expression of PD-L1 was assessed by immunohistochemistry. RESULTS: TILs status in CNB is significant in pathological therapeutic grade: 1 vs. 2 or 3 (p = 0.0359). Disease-free survival (DFS) in patients with Low-TIL tumors were significantly worse than those with High-TIL tumors (p = 0.0383), but overall survival (OS) showed no significance (p = 0.0772). However, in patients with Low-TIL tumors, both DFS and OS in patients with High-PD-L1 expression were extremely unfavorable than in patients with Low-PD-L1 expression (p = 0.0032, p = 0.0002). CONCLUSION: The patients with TNBCs with combined Low-TILs and High-PD-L1 status in pre-PST situation showed unfavorable prognosis. The subset of TNBCs with Low-TILs and High-PD-L1 status could be the therapeutic target for immune checkpoint inhibitor.

Voided urine cytology and low-grade urothelial neoplasia of the bladder: factors that influence the sensitivity.

INTRODUCTION: The aims of this study were to show the 10-year results of voided urine cytology (VUC) performed using liquid-based cytology (LBC) with CytoRich Red and to discuss the factors that influence the sensitivity of low-grade urothelial neoplasia (LGUN) of the urinary bladder. MATERIALS AND METHOD: We calculated the sensitivity of VUC in 421 histologically confirmed cases included in the pathology database of Hokkaido Cancer Center in Japan and studied various factors influencing sensitivity. RESULTS: The cumulative sensitivity of VUC was 95.8% in 143 cases of primary high-grade urothelial carcinomas, compared with 59.5% in 74 cases of LGUN. These findings were only slightly different from the previous results of Koss et al. The sensitivity in LGUN, however, showed lower values in some conditions, including in secondary cases, with a lower frequency of examinations and smaller tumor volumes. LBC preparations allowed us to observe a greater number of tumor cells and cell clusters than conventional methods in LGUN cases. CONCLUSIONS: The sensitivity of VUC can be improved by increasing the frequency of examinations and adopting a valid preparation method in order to augment the number of cells and cell clusters on individual glass slides. LBC preparations may allow cytopathologists to obtain a better sense for and understanding of the cytologic findings of LGUN.

Z-axis video for cytology database is a useful tool for the case presentation prior to the cytology training workshop.

We have built a database on the Internet managing z-axis video for cytology (Zavic), and report on a new style of case discussion supported by the Zavic database. Z-axis video for cytology is a movie file derived from the video recording of a microscopic field with changes in the focus. We used it for the case presentation of EUS-FNA of pancreatic lesions on the Internet prior to a training workshop. The attendees were asked to observe the Zavic and to make diagnoses of 20 cases before the workshop. Fourteen attendees also observed lesions under a microscope on that day, and the results were compared with those of Zavic observation. The evaluation of the Zavic database (DB) was surveyed by a questionnaire. The average number of accurate diagnoses by 46 Zavic observers was 10.8. These accuracies for those who observed both the Zavic and glass slides were 11.57 and 11.43, respectively, for the videos and slides. Compared with Zavic observation alone, the diagnoses with glass slide observation were shifted to a correct diagnosis in two cases, but were shifted to an incorrect diagnosis in two cases. Approximately 60% of Zavic observers replied in the questionnaire that the movies on Zavic DB started to play within 3-4 seconds after clicking the play button. We successfully carried out the new style of case discussion supported by the Zavic DB. It was evaluated favorably by many attendees, who were psychologically still dependent on the glass slide observation.

Ultrastaging of para-aortic lymph nodes in stage IIIC1 endometrial cancer: a preliminary report.

OBJECTIVE: The aim of this study was to determine the rate of occult metastasis, including isolated tumor cells, in para-aortic lymph nodes of patients with stage IIIC1 endometrial cancer who underwent pelvic and para-aortic lymphadenectomy. METHODS: A series of 15 patients who had undergone combined pelvic and para-aortic lymphadenectomy during the period from 2004 to 2010 and who were diagnosed as being positive for pelvic node metastasis but negative for para-aortic node metastasis were included in this study. Ultra-staging by multiple slicing, staining with hematoxylin/eosin and cytokeratin, and microscopic inspection was performed on a total of 242 para-aortic lymph nodes. RESULTS: Eleven (73.3%) of the 15 patients had occult para-aortic lymph node metastasis. Two patients (13.3%) had macrometastasis and nine patients (60.0%) had isolated tumor cells. Type 2 endometrial cancer tended to have a higher rate of occult metastasis than that of type 1 cancer (90% vs. 40%, P=0.07). The rate of occult para-aortic node metastasis was not related to the number of metastatic pelvic nodes. Five patients suffered recurrence in the lung or in the intraabdomen, but lymph node recurrence was not found in any case. CONCLUSION: Patients with stage IIIC1 endometrial cancer have a potentially high rate of occult para-aortic node metastasis. Local treatment of the para-aortic region should be considered in patients with stage IIIC1 endometrial cancer until effective adjuvant therapy is established.

Comparison between a traditional single still image and a multiframe video image along the z-axis of the same microscopic field of interest in cytology: Which does contribute to telecytology?

The limitation of cytologic still images is one of the reasons why telecytology has not met with widespread acceptance by the cytology community. Cytologic still image only displays a single depth of field, and this is a particularly acute problem in cytology where the specimen is often much thicker than a single microscopic depth of focus. In this article, we examine the validity of a "z-axis" video of a microscopic field of interest.After observing videos of fields of interest from 10 cases, five cytotechnologists reached suitable cytologic findings and diagnosed the fields correctly in great majority of cases. Five other cytotechnologists, who looked only at a single representative still image, could not always make a correct diagnosis. The difference between two observer groups was statistically significant by Wilcoxon's matched pairs signed-rank test.The results indicate that "z-axis" video of microscopic field of interest provides a similar experience to "focusing through" observation of the specimen under a microscope and may improve an accuracy of primary telecytodiagnosis. And we expect that video image telecytology will strongly influence cytology, especially in education and training.