Epigenetic modulation on sphere

Background : Cancer stem cell properties are highly relevant to the biology of treatment-resistant cancers. Epigenetic modification regulates gene expressions by chromatin remodeling during malignant transformation. The aim of this study was to elucidate the possible strategy for cancer stem cells focusing on epigenetic modification. Methods : We made cancer sphere from HepG2 cells, and we added Histone deacetylase (HDAC) inhibitor, valproic acid to cancer sphere. And we compared methylation status and the gene expression between normal HepG2 and cancer sphere groups, and between cancer sphere and sphere with HDAC inhibitor treatment groups. Results : Valproic acid (VPA) cancelled this spheroid formation. In comparison between normal HepG2 and cancer sphere, the number of methylation status changes more than 0.1 of beta level was 826 probes, and we could isolate some epithelial-mesenchymal transition (EMT) related genes. And VPA reduced the expressions of EMT related genes in sphere with RT-PCR. On the other hand, in comparison between cancer sphere and sphere with VPA treatment, we detected 29 probe of methylation status change, and VPA reduced the expressions of Bcl-6 in sphere. Conclusions : HDAC inhibitor affected the methylation status of cancer stem cells. Histone-acetylation might overcome treatmet-resistant cancer through the regulation of cancer stem cell

High expression of cancer stem cell markers in cholangiolocellular carcinoma

Purpose Cholangiolocellular carcinoma (CLC) is an extremely rare malignant liver tumor. It is thought to originate from the ductules and/or canals of Hering, where hepatic stem cells (HpSC) are located, but there are few reports on cancer stem cell markers in CLC. Thus, we evaluated the significance of cancer stem cell markers, including CD133, CD44, and EpCAM, in CLC. Methods The subjects of this study were three patients with CLC and one patient with an intermediate type of combined hepatocellular cholangiocarcinoma (CHC). The cancer cell markers, CK7, CK19, and EMA, were evaluated immunohistochemically. Results Histological examination of the CLC revealed morphologically cholangiolar features and immunohistochemical examination revealed positivity for CD133, CD44, and EpCAM. On the other hand, in the intermediate type of CHC, only CD44 was positive, whereas CD133 and EpCAM were negative. Conclusion CLC may have stronger features derived from HpSCs than an intermediate type of CHC

Treadmill Exercise as a Preventive Measure Against Age-Related Anxiety and Social Behavioral Disorders in Rats: When Is It Worth Starting?

Objective To determine the appropriate time points to start regular exercise which could reduce age-related anxiety and impaired social behavior. Methods For this study, 8-week-old male Wistar rats were divided into three groups: no exercise (NoEX), short-term exercise (S-Ex), and long-term exercise (L-Ex) groups. S-Ex-group rats started treadmill exercise at 12 months of age, while L-Ex rats started from at 2 months of age. Exercise rats were forced to walk on the treadmill three times per week, with 1- to 2-day intervals for 10 minutes during the first 2 weeks, at 10 m/min until 17 months of age, and at 8 m/min thereafter. At 19 months of age, behavioral tests were performed to assess the effects of exercise on age-induced behavioral change as well as quantitative polymerase chain reaction were done to uncover the mechanism behind the behavioral changes. Results Anxiety-like behavior was improved by long-term exercise. Additionally, rats belonging to the S-Ex and L-Ex groups showed improved social behavior and increased curiosity about interesting objects. The qPCR data showed that treadmill exercise suppressed the expression of immediate-early genes in the prefrontal cortex of the aged rats. Conclusion This study suggests that long-term exercise represses early response genes, and in this way, it increases resistance to stress, diminishes anxiety-related behavior, and improves social behavior. These findings underscore the need to consider appropriate time to start exercise to prevent stress induced anxiety related behavior

Hand-assisted laparoscopic splenectomy for a huge splenic vascular lesion with aneurysms in a patient with impending Kasabach-Merritt syndrome-like phenomenon

Splenic vascular lesions are relatively rare and are usually found incidentally. However, the vascular lesions associated with Kasabach-Merritt syndrome, such as hemangioma, can be life-threatening. We herein describe the case of a young adult female patient with a huge splenic vascular lesion, aneurysms of the splenic artery, and increased plasma levels of fibrin/fibrinogen degradation products and D-dimers. Hand-assisted laparoscopic splenectomy was performed, after which the coagulopathy was drastically improved. Minimally invasive surgical intervention such as hand-assisted laparoscopic splenectomy should be considered as the first treatment choice in such a case

Reduction of T-Box 15 gene expression in tumor tissue is a prognostic biomarker for patients with hepatocellular carcinoma

Genome-wide analysis is widely applied to detect molecular alterations during oncogenesis and tumor progression. We analyzed DNA methylation profiles of hepatocellular carcinoma (HCC), and investigated the clinical role of most heypermethylated of tumor, encodes T-box 15 (TBX15), which was originally involved in mesodermal differentiation. We conducted a genome-wide analysis of DNA methylation of tumor and non-tumor tissue of 15 patients with HCC, and revealed TBX15 was the most hypermethylated gene of tumor (Beta-value in tumor tissue = 0.52 compared with non-tumor tissue). Another validation set, which comprised 58 HCC with radical resection, was analyzed to investigate the relationships between tumor phenotype and TBX15 mRNA expression. TBX15 mRNA levels in tumor tissues were significantly lower compared with those of nontumor tissues (p < 0.0001). When we assigned a cutoff value = 0.5-fold, the overall survival 5-year survival rates of the low-expression group (n = 17) were significantly shorter compared with those of the high-expression group (n = 41) (43.3% vs. 86.2%, p = 0.001). Multivariate analysis identified low TBX15 expression as an independent prognostic factor for overall and disease-free survival. Therefore, genome-wide DNA methylation profiling indicates that hypermethylation and reduced expression of TBX15 in tumor tissue represents a potential biomarker for predicting poor survival of patients with HCC

ショウニ ノ タンカン カクチョウガタ スイ タンカン ゴウリュウ イジョウ セツジョレイ ニ ハツガン カンレン イデンシ HDAC ガ コウハツゲン シテイタ 1レイ

Pancreaticobiliary maljunction（PBM）is the congenital malformation that the junction of the pancreatic and bile ducts is located outside of the duodenal wall anatomically, so sphincter function preventing to mix the bile and pancreatic juice does not act. In PBM, the mutual reflux of bile and pancreatic juice induces various diseases and becomes the cause of biliary tract cancer generation. PBM is divided into PBM with the biliary dilatation and PBM without the biliary dilatation. From the perspective of carcinogenesis, surgery to separate the pancreatic juice and bile by excision of extrahepatic biliary tract and hepaticojejunostomy is commonly performed in the former, the treatment of the latter is controversial, for instance ; the only cholecystectomy should be performed and excision of extrahepatic biliary tract is unnecessary or excision of extrahepatic biliary tract and hepaticojejunostomy should be done. In addition, histone acetylation and histone deacetylation is one of the important regulatory mechanism of gene transcription, the error of the balance between histone acetyltransferase（HAT）and histone deacetylase（HDAC）leads to carcinogenesis. In other words, HDAC has a carcinogenic potential. Overexpression of HDAC was found in the both resected gallbladder and bile duct mucosa of infant PBM patient with the biliary dilatation performed the excision of extrahepatic biliary tract and hepaticojejunostomy in our department. Appropriate treatment for PBM has not been established yet, but even infant case with PBM has carcinogenesis potential just like this case. In conclusion, further accumulation of cases including PBM without the biliary dilatation was thought to be essential though the excision of extrahepatic biliary tract and hepaticojejunostomy still seems to be required for infant PBM patient with the biliary dilatation

ニクガンテキ モンミャク シンシュウ ヨウセイ カンガン セツジョゴ ノ Systemic IFN＋Low dose FP ノ ユウヨウセイ : リロンテキ コンキョ ト リンショウテキ コウカ

Background and Aims : Despite a recent progress of treatment for hepatocellular carcinoma （HCC）, the prognosis of advanced HCC with macroscopic vascular invasion remains unsatisfactory. We investigated anti-tumor effect of IFNα using experimental model and show the outcome of our systemic adjuvant therapy consisting of IFNα,５FU and cisplatin（IFP）after hepatectomy on advanced HCC with macroscopic portal invasion. Methods［: Basic study］Anti-tumor effects such as inhibition of invasion, proliferation of pegylated IFN α２b（PegIFNα）was evaluated using MH１３４mouse HCC cells, in vitro and vivo. ［Clinical study］: Thirty patients who had HCC with Vp２or more of macroscopic portal invasion（Vp２; portal vein tumor thrombus in its２nd order branch）were included. Those patients were retrospectively divided into two groups : the systemic IFNα,５FU and cisplatin group （n＝１４, IFP group）; and the no adjuvant therapy group（n＝１６, control）. Clinicopathological variables were compared between the two groups, including patient survival and disease-free survival. Results［: Basic］In vitro, the proliferation was significantly suppressed by Peg-IFNα, and invasion potential was also inhibited. In vivo, tumor growth was significantly suppressed compared to control （０．５vs.５．０cm, p＜０．０５）, and liver metastases was decreased（number :１９vs.６, p＜０．０５）. ［Clinical］The overall and disease-free survival rate in IFP group was significantly higher than in control group（１y :１００％ vs３８％,３y :６５％ vs２５％, P＜０．０１,１y :３６％ vs２５％,３y :３６％ vs１９％, P＜０．０１）. Regarding the recurrent patterns,５of９patients in IFP group had controllable tumors in the remnant liver, although１２of１３patients in control group had distant metastasis or multiple recurrences in the residual liver. Conclusion : Our new adjuvant regimen of systemic IFP may be a promising strategy after radical resection for HCC with macroscopic portal invasion

Effect of histone deacetylase inhibitor in combination with 5-fluorouracil on pancreas cancer and cholangiocarcinoma cell lines

Background : Histone deacetylase (HDAC) is well known to be associated with tumorigenesis through epigenetic regulation, and its inhibitors (HDACIs) induce differentiation and apoptosis of tumor cells. We examined the therapeutic effects of valproic acid (VPA, a HDACI) with a combination of 5-fluorouracil (5-FU) in vitro. Methods : A human pancreas cancer cell line (SUIT-2) and a cholangiocarcinoma cell line (HuCCT1) were used. Cell viabilities were evaluated by a cell proliferation assay. We determined the anticancer effects of VPA combined with 5-FU in these cell lines. Results : Pancreas cancer (SUIT-2) : No effect of 5-FU (1.0 μM) was observed, but 17% and 30% of proliferationinhibitory effects were recognized in a dose of 2.5 or 5.0 μM, respectively. Cell viability was only weakly reduced by VPA (0.5 mM). However, in combination of 5-FU (1.0 μM) with VPA (0.5 mM), 19% of inhibitory effect was observed. Cholangiocarcinoma (HuCCT1) : 5-FU (1.0 μM) did not suppress the cell viability, but 5-FU (2.5 μM) suppressed by 23%. VPA (0.5 mM) did not suppress the cell viability, while VPA (1.0 mM) weakly decreased it by 11%. Combination of 5-FU (1.0 μM) and VPA (0.5 mM) markedly reduced the cell viability by 30%. Conclusion : VPA augmented the anti-tumor effects of 5-FU in cancer cell lines. Therefore, a combination therapy of 5-FU plus VPA may be a promising therapeutic option for patients with pancreas cancer and cholangiocarcinoma