Transcriptome analysis of an AKT inhibitor-resistant endometrial cancer cell line.

BACKGROUND: Drug resistance in endometrial cancer (EC) is a serious problem and a barrier to improving prognosis. The PI3K/AKT/mTOR pathway is highly activated in EC and can serve as a potential therapeutic target. Inhibitors against AKT have been developed, but resistance to these inhibitors is a concern. This study aimed to establish AKT inhibitor resistant cell lines and identify differentially expressed genes (DEGs) between parental and AKT inhibitor resistant cell lines to understand the mechanism of drug resistance to AKT inhibitors in EC. METHODS: The sensitivity of eight EC cell lines to AKT inhibitor was analyzed. One of them was used to establish a drug-resistant cell line. DEGs were examined using RNA sequencing (RNA-seq). Furthermore, DEGs were comprehensively analyzed to identify hub genes. Hub genes were evaluated using quantitative real-time polymerase chain reaction. RESULTS: RNA-seq identified 617 DEGs. Hub genes were selected using bioinformatics analysis. The top 10 hub genes were TNF, CDH1, CCND1, COL1A1, CDH2, ICAM1, CAV1, THBS1, NCAM1, and CDKN2A. Relative mRNA expression was significantly upregulated for TNF, CDH1, CCND1, THBS1, p16INK4a, and p14ARF and significantly downregulated for CDH2, ICAM1, and NCAM1 in borussertib-resistant EC cell line. CONCLUSIONS: Drug resistance to AKT inhibitors may depend on genes related to cell adhesion-mediated resistance and transforming growth factor ß signaling.

Acquired AKT-inhibitor Resistance Is Mediated by ATP-binding Cassette Transporters in Endometrial Carcinoma.

BACKGROUND/AIM: Endometrial cancer is increasing in prevalence worldwide. It is treated with chemotherapy and radiotherapy, in addition to surgery, but the presence of treatment-resistant tumor cells remains a barrier to effective tumor control. The purpose of this study was to develop drug-resistant cell lines using triciribine, an AKT inhibitor, and investigate the mechanism of acquired resistance. MATERIALS AND METHODS: Triciribine sensitivity assays were performed using Cell Counting Kit-8 (CCK-8) on eight endometrial cancer cell lines. The chosen cell lines were highly sensitive to chemotherapy and radiotherapy. A new triciribine-resistant cell line was established and found to be highly resistant to chemotherapy. Properties of the resistant cell line were identified using molecular and cell biological techniques including CCK-8 and quantitative PCR analysis. RESULTS: HEC-151 had the highest triciribine sensitivity (IC50 value of 0.7±0.1 µM) of the endometrial cancer cell lines tested. We established a triciribine-resistant cell line from HEC-151 by growing cells in the presence of increasing concentrations of triciribine up to 66.6 µM. The resistant HEC-151 cells changed to spindle-shaped morphology and importantly reduced triciribine sensitivity compared to the parental cell line. ABC transporters involved in drug efflux had significantly higher expression levels in ABCB1 (1.4±0.10 times higher), ABCC1 (11.4±0.22 times higher), and ABCC4 (4.5±0.42 times higher). CONCLUSION: In this study, we established a triciribine-resistant cell line from HEC-151 cells. Our data suggest that the mechanism of drug resistance in endometrial cancer cells is attributed to the increased expression of ABC transporters.

[A Case of Advanced Adenocarcinoma at the Esophagogastric Junction That Exhibited pCR by Preoperative SOX Therapy].

Although neoadjuvant chemotherapy(NAC)is an effective treatment option for advanced adenocarcinoma at the esophagogastric junction (AEG), there is no sufficient evidence of this in Japan. We report a case of advanced AEG with pathological complete response(pCR)after NAC with S-1 and oxaliplatin(SOX). A 39-year-old man was diagnosed with advanced AEG cT3(SS)N0M0, cStage ⅡB. A total of 3 courses of SOX was administered. After the chemotherapy, the primary tumor showed a significant reduction in size. Subsequently, laparoscopic proximal gastrectomy, D1+ lymphadenectomy and double-flap technique reconstruction were performed. Histopathological examinations showed no residual cancer cells in the resected specimen. Thus, preoperative SOX therapy can be one of the useful treatment strategies for advanced AEG.

Diagnostic significance of apical membranous and cytoplasmic dot-like CD26 expression in encapsulated follicular variant of papillary thyroid carcinoma: a useful marker for capsular invasion.

Non-invasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) and invasive encapsulated follicular variant of papillary thyroid carcinoma (EFV-PTC) are indistinguishable preoperatively. CD26 expression in follicular tumor-uncertain malignant potential (FT-UMP) is reported to be clearly higher than in that without capsular invasion. To verify the diagnostic significance of CD26 immunostaining in EFV-PTC, we examined the expression pattern of CD26 in non-invasive EFV-PTC (NIFTP) and invasive EFV-PTC. We performed immunohistochemical analysis using CD26 antibody for 37 NIFTPs and 54 EFV-PTCs (34 minimally invasive EFV-PTCs and 20 widely invasive EFV-PTCs). Most NIFTP samples showed an apical membranous pattern or a cytoplasmic diffuse pattern of expression. Invasive EFV-PTCs more frequently showed a cytoplasmic dot-like pattern, and the labeling indices of tumor cells with cytoplasmic dot-like patterns were significantly higher than those in NIFTPs. The sizes of dots seen in NIFTPs (mean: 1.12 µm) were significantly smaller than in invasive EFV-PTCs (1.33 µm), minimally invasive EFV-PTC (1.27 µm), and widely invasive EFV-PTC (1.38 µm). We, therefore, conclude that cytoplasmic diffuse and/or cytoplasmic dot-like CD26 expression, particularly the larger CD26-positive dots, could be useful markers for capsular invasion in EFV-PTC. CD26 immunostaining, using cell blocks or cytological specimens, may preoperatively distinguish between NIFTP and invasive EFV-PTC.

Diagnostic and therapeutic role of endoscopic retrograde pancreatography and stent placement for grade IV blunt pancreatic trauma: A case report.

Pancreatic trauma involving ductal injury is rare but is associated with high morbidity and mortality. The benefit of endoscopic retrograde pancreatography and stent placement is unclear because there are only a few case reports on endoscopically treated pancreatic duct transection at the pancreatic head. We report a rare case of grade IV pancreatic trauma successfully treated with endoscopic pancreatic stent, which we believe makes significant contribution to the existing literature. A 17-year-old man with blunt pancreatic trauma was referred to our hospital and was diagnosed with grade IV pancreatic injury using endoscopic retrograde pancreatography. The patient was successfully managed with endoscopic pancreatic duct stenting. Although stent replacement was required three times and a trivial ductal stricture remained, the patient finally became stent-free without any symptoms and further adverse events. Endoscopic retrograde pancreatography is highly advantageous for early detection and evaluation of the severity of ductal injury. Subsequent stent insertion is well tolerated in hemodynamically stable patients and is especially beneficial for the treatment of pancreatic head injuries because it allows avoidance of sub-total pancreatectomy or high-risk reconstructive surgery. Nevertheless, the long-term outcomes and appropriate management of main pancreatic duct strictures due to stents remain to be determined. Accumulation of similar case experiences is essential to address these issues.

Proliferation of poorly differentiated endometrial cancer cells through autocrine activation of FGF receptor and HES1 expression.

Patients with poorly differentiated endometrial cancer show poor prognosis, and effective molecular target-based therapies are needed. Endometrial cancer cells proliferate depending on the activation of HES1 (hairy and enhancer of split-1), which is induced by several pathways, such as the Notch and fibroblast growth factor receptor (FGFR) signaling pathways. In addition, aberrant, ligand-free activation of the FGFR signaling pathway resulting from mutations in FGFR2 was also reported in endometrial cancer. However, a clinical trial showed that there was no difference in the effectiveness of FGFR inhibitors between patients with and without the FGFR2 mutation, suggesting a presence of another signaling pathway for the FGFR activation. Here, we investigated the signaling pathway regulating the expression of HES1 and proliferation of poorly and well-differentiated endometrial cancer cell lines Ishikawa and HEC-50B, respectively. Whereas Ishikawa cells proliferated and expressed HES1 in a Notch signaling-dependent manner, Notch signaling was not involved in HES1 and proliferation of HEC-50B cells. The FGFR inhibitor, NVP-BGJ398, decreased HES1 expression and proliferation of HEC-50B cells; however, HEC50B cells had no mutations in the FGFR2 gene. Instead, HEC-50B cells highly expressed ligands for FGFR2, suggesting that FGFR2 is activated by an autocrine manner, not by ligand-free activation. This autocrine pathway activated Akt downstream of FGFR for cell proliferation. Our findings suggest the usefulness of HES1 as a marker for the proliferation signaling and that FGFR inhibitor may be effective for poorly differentiated endometrial cancers that harbor wild-type FGFR.

[A Case of Carcinoma Associated with an Anal Fistula Repaired Using a Hatchet Flap after Neoadjuvant Chemoradiotherapy].

The patient was a 53-year-old man who complained of perianal pain. He had a 20-year history of an anal fistula. There was a 10 cm tumor with mucin-like discharge on the left side of the anus. His CEA level was high, at 7.3 ng/mL, and T2-weighted MRI revealed a multilocular cystic tumor with high signal intensity. The second biopsy result indicated mucinous adenocarcinoma. We performed neoadjuvant chemoradiotherapy. The chemotherapy regimen was TS-1 with radiation therapy, along with 4-port irradiation(50.4 Gy)of the primary tumor, located at the interior of the pelvis and inguinal lymph nodes. At the time of treatment completion, a partial response(PR)was observed, and a complete response(CR)was obtained after 6 months. We performed laparoscopic abdominoperineal resection. The large defect in the perianal skin and the pelvic cavity was repaired using a hatchet flap. The final diagnosis was confirmed as mucinous adenocarcinoma, pT4b, ly0, v0, N0, pPM0, pDM0, pRM0, pStage II . The patient was discharged 20 days after surgery. There is no indication of recurrence of the cancer after 1 year, and he continues to visit the outpatient clinic for regular follow-ups. Neoadjuvant chemoradiotherapy was effective in this case.

Coordination Structure Conversion of Hydrazone-Palladium(II) Complexes in the Solid State and in Solution.

We prepared hydrazone-palladium(II) complexes of [PdCl2(HL(n))] and [PdCl(L(n))] (n = 1-3) by the reaction of [PdCl2(cod)] or [PdCl2(PhCN)2] and the hydrazone ligands of HL(n) {N'-(pyridin-2-ylmethylene)picolinohydrazide (HL(1)), N'-[1-(pyridin-2-yl)ethylidene]picolinohydrazide (HL(2)), and N'-[(6-methylpyridin-2-yl)methylene]picolinohydrazide (HL(3))}. The structures of the complexes were determined by X-ray analysis. The hydrazone ligands had κN(py1),κN(imine) and κN(amidate),κN(py2) bidentate coordination modes in [PdCl2(HL(n))] (1, n = 1; 2, n = 2) and in [PdCl2(HL(3))] (3), respectively. In contrast, tridentate coordination modes of κN(py1),κN(imine),κN(py2) and κN(py1),κN(amidate),κN(py2) were observed in [PdCl(L(n))] (4, n = 1; 5, n = 2) and in [PdCl(L(n))] (6, n = 1; 7, n = 2; 8, n = 3). Thermal conversion of complexes 1-3 to complexes 6-8 proceeded in acetonitrile. Complexes 4 and 5 were obtained from complexes 1 and 2, respectively, in a basic acetonitrile solution under dark conditions. Complex 4 reverted immediately to complex 1 in an acidic acetonitrile solution that included hydrochloric acid. However, under room light, in the basic acetonitrile solution that included trimethylamine, complex 4 converted photochemically to complex 6. The thermochromic or vapochromic structure conversion of these complexes also occurred in the solid state. On heating at 180 °C, the color of the crystals of complexes 1, 2, and 3 changed from yellow to orange in the solid state. (1)H NMR and/or UV-vis absorption spectroscopy confirmed that the orange complexes 6-8 were produced. The reddish-orange crystals of complexes 4 and 5 were exposed to hydrogen chloride vapor to yield the yellow products of complexes 1 and 2, respectively.

[Two cases of dural metastasis of breast cancer].

The first case of dural metastasis occurred in a 60s years old woman, who presented with bone metastasis to the right breast. Nine months later, disorientation and left hemiplegia developed, the right coronal bone metastasis enlarged, and dural metastases were detected close to the tumor, as observed by using cranial magnetic resonance imaging (MRI). Whole brain radiation and chemotherapy(weekly paclitaxel)were administered. The right coronal bone metastasis reduced remarkably, and the dural metastases almost disappeared, as observed on a cranial MRI scan. The second case of dural metastasis occurred in a 50s years old woman who presented with multiple bone metastases. Extensive bone metastases to the skull and dural metastases to the side of the head were observed on cranial MRI scans. Subsequently, the patient experienced a severe headache, and whole brain radiation and pharmacotherapy with anastrozole and trastuzumab were administered. Cranial MRI revealed that the skull bone metastasis reduced and the dural metastases almost disappeared. We report that radiotherapy and pharmacotherapy were effective in these 2 cases of dural metastases of breast cancer.

[A case of a geriatric patient with stage IV anal canal cancer showing complete response to chemoradiation therapy].

We present a case in which chemoradiation therapy was effective in a geriatric patient with Stage IV anal canal cancer. The patient is an 81-year-old woman who complained of proctorrhagia and anal pain. She was referred to us by her family doctor who suspected rectal cancer. Tumors as large as 6.5 cm in diameter mainly on the right side of the rectum as well as 2 palpable enlarged lymph nodes on the right inguinal area, were found during the initial physical examination. Squamous cell carcinoma was elevated to 16 ng/mL. A CT scan revealed that irregularly shaped masses as large as 7 cm in diameter were externally exposed on the right side of the rectum along with enlarged lymph nodes on the right inguinal area and metastasis at S7 lesion in the liver. Squamous cell carcinoma was diagnosed from biopsy results. Due to her age, the chemotherapy regimen was S-1+CDDP with radiation therapy and 4-port irradiation (50.4 Gy) of the primary tumor, interior of the pelvis, and inguinal lymph nodes. Partial response was observed upon completion of treatment, and complete response was obtained after 6 months. She is currently an outpatient taking S-1: 60 mg/day orally. There is no indication of cancer recurrence after 1 year and 3 months, and she continues to visit an outpatient clinic for regular follow-ups. These results demonstrate the effectiveness of chemoradiation therapy for geriatric patients with Stage IV anal canal cancer.

Abscess formation of the round ligament of the liver: report of a case.

Abscess formation of the round ligament of the liver is very rare. We report a case of a 70-year-old female with abscess of the round ligament after an endoscopic papillotomy for choledocholithiasis. On the 21st day following papillotomy, abscess formation of the round ligament was found by ultrasonographic examination. Surgical treatment was performed because conservative therapy was not effective. The purulent fluid and necrotic tissue at the round ligament were completely removed. Cultures obtained from the abscess grew Staphylococcus epidermidis, but the mechanism of abscess formation in this case remains unclear.

The outcome of gastrointestinal stromal tumors (GISTs) after a surgical resection in our institute.

PURPOSE: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract arising from Cajal's cells and expressing c-kit. In a consensus report, the clinical behavior of GISTs was categorized into risk classes according to the tumor size and mitotic count. We analyzed the risk categories based on GIST patients who underwent a surgical resection at our institute during a period of 15 years. METHODS: We evaluated the risk categories of GISTs and analyzed the outcome and risk categories retrospectively. We presented the MIB-1 score of the tumor instead of mitotic counts for the evaluation of cellular growth because of inaccuracies regarding the mitotic counts. RESULTS: Patients were classified into 4 cases of very low risk, 11 of low risk, 8 of intermediate risk, and 5 of high risk. Four high-risk patients showed recurrence as either liver metastasis or peritoneal dissemination. In addition, local recurrence occurred in one low-risk and one intermediate-risk patient each. CONCLUSION: Our cases confirmed the correlation between the risk categories and the prognosis. A complete resection with sufficient margin must be confirmed even in low-risk cases to prevent local recurrence. Since high-risk patients showed poor prognosis, the adjuvant treatment with chemotherapeutic regimens must therefore be further studied for high-risk patients.

Multiple gastrointestinal stromal tumors in neurofibromatosis type 1 treated with laparoscopic surgery.

Gastrointestinal stromal tumors (GISTs) have been reported to occasionally occur in patients with neurofibromatosis type 1 (NF-1), and many cases have had multiple lesions predominantly involving the small intestine. We report herein a case of multiple GISTs associated with NF-1 from whom laparoscopic surgery was beneficial. In a 79-year-old female admitted with anemia and melena, the abdominal computed tomography revealed a tumor arising from the small intestine. Laparoscopic surgery was performed, and another small tumor was revealed during laparoscopic observation. Extracorporeal partial and wedge resection of the small intestine were undertaken. Both lesions were diagnosed as typical GISTs of low risk. Laparoscopic surgery would be useful for examination and a minimally invasive approach to tumors of the small intestine, especially on cases with the possibility of multiple tumors.

Abdominal wall and thigh abscess resulting from the penetration of ascending colon cancer.

An unusual case is described in which an abdominal wall and thigh abscess was an initial symptom of ascending colon cancer. A 76-year-old woman was referred to our hospital for investigation of fever and abdominal and thigh swelling. Computed tomography revealed a right abdominal wall, retroperitoneal, psoas and thigh abscess formation suspected to be caused by colon perforation. Due to the patient's poor general condition, local drainage of the abscess was performed on the following day of hospitalization. Histological examination of necrotic tissues removed form the retroperitoneal cavity demonstrated adenocarcinoma of the colon. The patient subsequently underwent right hemicolectomy with lymph nodal dissection after 19 days of the drainage procedure and was transferred to another hospital on the 49th day following the second surgery.