A Rare Case of Pleural Epithelioid Mesothelioma With a Prominent Myxoid Stroma Reported With Morphology, Fluorescent In Situ Hybridization, and Ultrastructural Findings.

Herein, we report a rare case of pleural epithelioid malignant mesothelioma with a prominent myxoid stroma. To date, detailed morphological or molecular pathological findings have not been reported for this type of tumor. Hence, we aimed to describe the cytological, histological, immuno-cytohistological, electron-microscopic, and molecular pathological findings using fluorescence in situ hybridization (FISH) in such a case. The patient was a male in his mid-sixties with a history of asbestos exposure and had originally visited the hospital with a persistent cough and fever. Chest radiography revealed left pleural effusion, and laboratory examination revealed a high titer for hyaluronic acid in the effusion. Additionally, computed tomography revealed diffuse multinodular or cystic lesions in the left parietal pleura, and pleural effusion cytology revealed large epithelioid cells with mild nuclear atypia, which were considered reactive mesothelial cells. Cytologically, Giemsa staining revealed that these cells harbored variously sized intracytoplasmic vacuoles that were Alcian-blue-positive, suggesting hyaluronan production. Biopsy revealed large epithelioid cells that loosely proliferated against a prominent myxoid background. These cells were immuno-positive for calretinin, Wilms' tumor 1, D2-40, vimentin, and cytokeratin AE1/AE3 but not for carcinoembryonic antigen, Ber-EP4, or desmin. BRCA 1 associated protein 1 immunostaining showed nuclear loss, and FISH showed homozygous deletion of cyclin-dependent kinase inhibitor 2A (p16) on chromosome 9p21. Based on these findings, the lesion was diagnosed as an epithelioid mesothelioma with a prominent myxoid stroma. Electron-microscopy demonstrated a dense microvillus pattern on the surface of the tumor cells, indicating a mesothelial cell origin, and variously sized vacuoles in the cytoplasm, confirming the presence of intracytoplasmic vacuoles demonstrated on cytology. The tumor tissues obtained during surgery harbored prominent myxoid stroma, which proved that the present tumor was consistent with this type of mesothelioma. After informed consent was obtained, the patient and family wished for total resection of the tumor and postoperative chemotherapy, and the patient eventually died eight months after surgery.

[Thoracoscopic Right S6b+S8a+S9a Combined Subsegmentectomy for Small-sized Lung Cancer].

A 76-year-old man visited to the hospital with a complaint of cough. The chest computed tomography scan revealed a 20 mm part-solid nodule in the right lower lobe, which was suspected to be an early-stage lung cancer. Since the tumor was located in S8a, and was adjacent to S6b and S9a, thoracoscopic right S 6b+S 8a+S 9a combined subsegmentectomy was performed. Virtual-assisted lung mapping (VAL-MAP) with indocyanine green( ICG) was performed as the preoperative marking. After dissecting the target vessels and bronchi, the intersegmental plane was identified using intravenous ICG. The pathological diagnosis was invasive mucinous adenocarcinoma with no lymph node metastasis. Combined subsegmentectomy is a useful option for the sublober resection of small lung cancer.

Migration of a hookwire used as a video-assisted thoracoscopic surgery marker into the splenic artery.

We present a case in which a hookwire that was used as a video-assisted thoracoscopic (VATS) surgery marker migrated into the splenic artery. The patient was a 70-year-old man with an 18-mm ground glass nodule (GGN) in the right S2. As the GGN was not located in the peripheral part of the lung, a percutaneous hookwire was placed as a marker under CT-guided just before the surgery. We performed VATS right S2 segmentectomy to remove the GGN and the marker; however, we could not locate the marker in the specimen. Histopathological examination revealed adenocarcinoma, TisN0M0, stage 0. CT findings after surgery showed that the marker had migrated into the splenic artery. We followed up the patient, and CT examination conducted 1, 3 and 6 months after the surgery showed no further migration and no damage of the splenic artery. We report the complication of percutaneous hookwire migration into a blood vessel.

[A Case of Anal Canal Squamous Cell Carcinoma Detected with Anal Stenosis Eight Years Following Resection for Rectal Adenocarcinoma].

An 89-year-old woman underwent low anterior resection for rectal adenocarcinoma(Ra, pT3N0M0, pStage II , Cur A)in 2008. In February 2016, she underwent an outpatient examination because of a defecation disturbance. Lower gastrointestinal endoscopy was performed and the stenotic region was biopsied. However, no malignancy was detected and the stenotic site expanded. However, the patient experienced recurrence of the same symptoms, developed severe anal stenosis, and underwent another examination in December 2016. Magnetic resonance imaging indicated a neoplastic lesion around the entire circumference of the anal canal. Transperineal needle biopsy results indicated squamous cell carcinoma. The patient was diagnosed with postoperative rectal cancer and metachronous anal canal squamous cell carcinoma(P, cT4bN2M0, cStage III b). Laparoscopic artificial anus construction was performed with the aim of unblocking the anal canal stenosis. Considering the patient's age and performance status, radiation therapy was administered. Two months after administering radiation therapy, the tumor decreased in size, and anal pain reduced.

[Clinical experience of the articulating needle-holder for completely video-assisted thoracoscopic surgery].

The autonomy Laparo-angle needle holder is a flexible device which has several articulating parts facilitating some traditionally difficult way of suture passage. This device is often used for laparoscopic surgery, and there have been few reports for video-assisted thoracic surgery (VATS). We used this device for complete VATS lobectomy and segmentectomy, and it enables us to suture a bronchus with the optimal direction even in the deep surgical field on complete VATS lobectomy and segmentectomy. Although some training may be needed to freely manipulate this device, it can be useful for minimallyinvasive video-assisted thoracic surgery.

Antitumor effects and drug interactions of the proteasome inhibitor bortezomib (PS341) in gastric cancer cells.

The proteasome inhibitor bortezomib (PS341) inhibits the function of the 26S proteasome and has been extensively investigated in the clinical setting of hematologic malignancies. Remarkable efficacy has been reported in the treatment of multiple myeloma, but there have been few studies of its use in the treatment of gastrointestinal malignancy, especially gastric cancer. Here, we demonstrate its efficacy, both alone and in combination with other cytotoxic agents, in gastric cancer cell lines. The human gastric cancer cell lines AZ521, MKN45 and NUGC3 were used as experimental models. Bortezomib produced significant growth inhibition in these cells (mean IC50 values: 1.26, 9.44 and 8.63 micromol/l, respectively) and was also observed to decrease the activity of the extracellular signal-regulated kinase 1/2 and Akt signal pathways, increasing the accumulation of p21. Cell-cycle analysis revealed that a low concentration of bortezomib (10-100 nmol/l) increased accumulation in the G1 phase. Moreover, bortezomib showed synergistic growth inhibition in combination with the conventional cytotoxic agents 5-fluorouracil, paclitaxel, doxorubicin and SN-38, and also downregulates the activity of nuclear factor -kappaB, which is induced by these agents. Our results demonstrate that bortezomib could be an effective antitumor agent in the treatment of gastric cancer, both as single-agent therapy and in combination with conventional chemotherapeutic agents.

Proteasome inhibitor bortezomib increases PTEN expression and enhances trastuzumab-induced growth inhibition in trastuzumab-resistant cells.

PTEN (phosphatase and tension homolog deleted on chromosome 10) has been shown to be inactivated in a wide range of cancers and the role of this gene product is associated with the suppression of the phosphatidylinositol-3-kinase (PI3K)/Akt pathway in many cancers. Recently, some reports demonstrated that the degree of PTEN expression could predict trastuzumab chemosensitivity in ErbB2-overexpressing breast cancer. Here, we demonstrate the possible involvement of a proteasome inhibitor (PS341) in PTEN expression and elucidate the influence of PI3K/Akt, one of the main cascades of the ErbB2 downstream pathway, and discuss the role of the proteasome inhibitors in trastuzumab resistance. ErbB2-overexpressing SKBR3 human breast cancer cells and trastuzumab-resistant SKBR3/R cells were analyzed in this study. We show that the expression of phosphorylated Akt was highly increased in trastuzumab-resistant cells, although the expression of PI3K, phosphorylated PI3K and non-phosphorylated Akt was unchanged in comparison with wild-type SKBR3 cells. However, following treatment with PS341, the level of phosphorylated Akt was decreased in a dose-dependent manner. Conversely, the level of PTEN was increased in the same fashion. PS341 showed sufficient cytotoxicity in resistant cells in combination with trastuzumab and the efficacy of trastuzumab was inclined to be better in resistant cells under PS341 treatment. Remarkable activity of Akt was observed in trastuzumab-resistant SKBR3 breast cancer cells and this phenomenon could be associated with the decreased expression of PTEN. The proteasome inhibitor PS341 could increase the level of PTEN and inhibit the downstream pathway of ErbB2, interfering with phosphorylation of Akt.

Proteasome inhibitors can alter the signaling pathways and attenuate the P-glycoprotein-mediated multidrug resistance.

Numerous signaling pathways were reported to be involved in the resistance for conventional cytotoxic drugs, although one of the main reasons is the overexpression of P-glycoprotein (P-gp) in multidrug resistant cancer cells. The overexpression of P-gp has been associated with the resistance to a wide range of anticancer drugs. Doxorubicin and paclitaxel are substrates of this transporter system and have an important role for the various human malignancies. In the present study, drug-sensitive MCF7 and multidrug resistant MCF7/ADR (characterized by overexpression of P-gp) human breast cancer cell lines were used as an experimental model. We have found that PS341 and MG132, proteasome inhibitors, reduced the degree of the multidrug resistance (MDR) in MCF7/ADR cells. This phenomenon was accompanied by a decrease in the IC50 value of doxorubicin and paclitaxel from 55.9 +/- 3.46 to 0.60 +/- 0.08 microM, and from 17.61 +/- 1.77 to 0.59 +/- 0.12 microM, respectively. The IC50 values of sensitive cells for doxorubicin and paclitaxel were about 0.42 and 0.83 microM, respectively. The effect of PS341 and MG132 on MCF7/ADR cells was associated with a significant decrease in both protein and gene levels of P-gp expression. Moreover, with regard to the expression of possible signal transduction pathways of mitogen-activated protein kinase (MAPK) related to the activation of mdr1, proteasome inhibitors did significantly influence the activation of these proteins. Western blot analysis revealed that 24 hr exposure of multidrug resistant MCF7/ADR cells with proteasome inhibitors did change the levels of DNA binding activity of nuclear factor-kappaB (NF-kappaB), pERK1/2, c-Jun, and p-c-Jun. In conclusion, we could remark that proteasome inhibitors (especially PS341) attenuate the resistance of MCF7/ADR cells for P-gp substrate drugs of doxorubicin and paclitaxel. Several proteins are supposed to be associated with the resensitization of the cells to conventional cytotoxic drugs, although decreased activity of P-gp is at least involved in the proteasome inhibitor-related resensitization. And influence with MAPK pathways, which have been reported to be associated with the regulation of P-gp, might be contributed to the resensitization brought by proteasome inhibitors.

The relationship between NY-ESO-1 mRNA expression and clinicopathological features in non-small cell lung cancer.

Tumor-associated antigens recognized by cellular or humoral effectors of the immune system are potential targets for antigen-specific cancer immunotherapy. NY-ESO-1 is one of the most immunogenic cancer/testis (CT) antigens and emerges as the potential candidate for specific immunotherapy. We studied mRNA expression status of NY-ESO-1 in 63 cases of NSCLCs using the real-time reverse transcription PCR to examine the relationship between its expression and clinicopathological features. NY-ESO-1 expression was present in 20 (32%) of 63 NSCLC cases and significantly increased with the advancement of disease stage in TNM classification (P=0.013), especially related to lymph node metastasis (P=0.020). Moreover, frequency of NY-ESO-1 expression was related to the degree of pathological differentiation (P=0.035). The quantity of NY-ESO-1 expression by real-time RT-PCR was not correlated with any clinicopathological factor. Our results demonstrate that the NY-ESO-1 expression was frequently present in primary NSCLC, especially advanced cases with lymph node metastasis. In addition, the high incidence of NY-ESO-1 expression in NSCLC suggests the possibility of a specific immunotherapy for NSCLC.

The effect of epidermal growth factor receptor antisense morpholino oligomer on non-small cell lung cancer cell line.

Overexpression of the epidermal growth factor receptor (EGFR) has been identified as a common component of various cancer types including lung cancer. Recently morpholino oligonucleotides appeared as a promising modification for antisense applications with few toxic effects and their stability. We investigated the effect of EGFR antisense morpholino oligomer on non-small cell lung cancer (NSCLC) cell line by evaluating EGFR mRNA, protein product and cell proliferation. The EGFR antisense morpholino oligomer was designed to target the translation start site in the EGFR mRNA. The four base-mismatch morphlino oligomer was designed as a control for EGFR antisense morpholino oligomer. These morpholino oligomers were introduced into NCI-H125 cell line which showed overexpression of EGFR. The EGFR mRNA and protein expression were quantified by real time RT-PCR and ELISA, respectively. The significant repression in both EGFR mRNA and protein expression was observed for three days after single treatment with EGFR antisense morpholino oligomer. Furthermore, the growth of NCI-H125 cell line was significantly inhibited with treatment by EGFR antisense morpholino oligomer. Our results indicate that EGFR antisense morpholino oligomer represses the EGFR expression at both mRNA and protein level and inhibits the proliferation of NSCLC cell line suggesting that it may be a promising strategy as one of antisense therapies for NSCLC.

Mediastinal parathyroid cyst with tracheal constriction.

A 63-year-old man visiting a physician for slight dyspnea, attributed to a lump on his neck, was found in ultrasonography and computed tomography to have a cyst extending from the left lobe of the thyroid gland to the superior mediastinum. Radiography showed right deviation of the trachea. The cyst disappeared after fine-needle aspiration, but cyst fluid subsequently reaccumulated and he was admitted to our hospital. No abnormalities were detected in tests of thyroid and parathyroid function or blood chemical analysis. The cyst was surgically removed and diagnosed as a nonfunctioning parathyroid cyst, based on the high-intact parathyroid hormone in cyst fluid. The patient recovered fully and has shown no recurrence in the 11 months to data since surgery.

Enhancement of gene transduction efficiency in cancer cells using cationic liposome with hyperthermia.

We evaluated the effects of hyperthermia on the efficiency of gene transduction by using a cationic liposome to develop an efficient method for lipofection. We used Lewis lung carcinoma (LLC), NIH3T3, and A549 cell lines, with Lipofectamine reagent as the cationic liposome and the LacZ gene as the reporter gene. In LLC, co-incubation of the cationic liposome and plasmid DNA complex (lipoplex) with the cells for 2 h at 41 degrees C enhanced the efficiency of gene transduction approximately 1.4-fold compared to incubation for 2 h at 37 degrees C, as measured by X-gal staining and beta-galactosidase activity. In cell lines NIH3T3 and A549, the efficiency of gene transduction showed a tendency toward enhancement after 2 h co-incubation with lipoplex at 41 degrees C compared to that at 37 degrees C, as measured by X-gal staining. This is the first study to demonstrate the enhancement of gene transduction efficiency achieved by using a cationic liposome under conditions of hyperthermia. This method should prove useful for lipofection in other cancer cells.