Epithelial-to-Mesenchymal Transition in Pancreatic Adenocarcinoma

Epithelial to mesenchymal transition (EMT) is a physiologic process that allows morphological and genetic changes of carcinoma cells from an epithelial to a mesenchymal phenotype, which is the basis of the high metastatic potential of pancreatic cancer cells. EMT is triggered by various tumor microenvironmental factors, including cytokines, growth factors, and chemotherapeutic agents. This review summarizes the state-of-the-art knowledge on the molecular mechanisms that support pancreatic cancer EMT and the evidences that support its involvement in invasiveness/aggressiveness, and the drug resistance of pancreatic cancer cells

慢性膵炎における膵炎関連蛋白の発現とアポトーシスの意義に関する実験的・臨床的研究

金沢大学がん研究所ラット自然発症慢性膵炎モデル(WBN/Kob)では12週令から慢性膵炎の変化が起こり,週令とともに顕著になった.RT-PCR法による膵組織中pancreatitis-associated-protein(PAP)mRNAの発現動態の検討では,膵炎発症前の8週令から上昇し,12週令でピークに達し,24週令でほとんど消失した.PAPが膵腺房細胞に発現することをmRNAレベルではin situ hybridization法,蛋白レベルでは免疫染色法により確認した.膵炎治療薬(メシル酸カモスタット,柴胡桂枝湯)投与により,膵炎の発症と進展が抑制され,PAPmRNA発現も低下した.アポトーシスの検討では,膵内炎症細胞にアポトーシス陽性細胞が認められたが,膵腺房細胞にはほとんど検出されなかった.膵腺房培養細胞を用いたin vitroの系で,PAPのアポトーシス抑制作用を見い出し,現在さらに検討中である.臨床的には,196例の消化器疾患において血清PAP値をELISA法により測定したところ,その陽性率は急性膵炎で90%であり,膵炎の重症度と有意に関連し,その上昇は他の膵酵素に比し遷延し,急性膵炎の治癒に一致して正常化した.また,各種消化器癌での血清PAPの陽性率は,胃癌16%,大腸癌40%,肝細胞癌40%,胆道癌26%,膵癌40%であった.以上より,慢性膵炎の発症と進展にPAPの発現が関連し,その意義としてアポトーシス抑制作用が考えられた.また,血清PAP値が膵炎の重症度や治癒判定の指標になること,癌細胞での異所性発現を反映して消化器癌の一部でも血清PAP値が上昇することを明らかにした.In spontaneous chronic pancreatitis model (WBN/Kob rats), pancreatitis was first observed at 12 weeks, and progressed thereafter. By the analysis with RT-PCR, PAP mRNA started to be expressed at 8 weeks, peaked at 12 weeks, and then almost disappeard at 24 weeks. PAP was expressed in the cytoplasm of pancreatic acinar cells by the analysis with in situ hybridization and immunohistochemistry. Expression of cytokines such as TNF-alpha and IL-6 was also studied. TNF-alpha and IL-6 peaked at 8 and 12 weeks, respectively. Drug therapy (camostat mesilate and Saiko-keishi-to) prevented the development of pancreatitis and suppressed the gene expression of PAP and cytokines.As for apoptosis, the TUNEL method revealed abundant apoptotic inflammatory cells in the stroma of the pancreata of WBN/Kob rats at 12 weeks. However, only a few apoptotic cells were detected in the acini. Anti-apoptotic effets of PAP were suggested in in vitro system using a rat pancreatic acinar AR42J cell line treated with L-arginine.Clinical studies on serum PAP in 196 patients with digestive diseases showed that serum PAP was increased in 90% cases of acute pancreatitis, which was significantly higher than the positive rate of 9% in clinically stable chronic pancreatitis. Serum PAP levels were significantly related to the severity of pancreatitis. The elevation of serum PAP prolonged compared to other pancreatic enzymes or CRP, and serum PAP was normalized at the time of clinical healing of acute pancreatitis. The positive rates of serum PAP in gastrointestinal cancers were as follows : gastric cancer 16%, colorectal cancer 40%, hepatocellular carcinoma 40%, biliary tract cancer 26% and pancreatic cancer 40%. Serum PAP was normalized after curative resection of the tumor in 7 cases.These results suggest that PAP expression is related to the development and progression of chronic pancreatitis and that the action mechanisms of PAP include anti-apoptotic effects. Clinical studies implicate that serum PAP is a useful marker of the severity and healing of acute pancreatitis and that serum PAP is elevated in in cancer cells.研究課題/領域番号:09670523, 研究期間(年度):1997 – 1998出典：研究課題「慢性膵炎における膵炎関連蛋白の発現とアポトーシスの意義に関する実験的・臨床的研究 」課題番号09670523（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-09670523/096705231998kenkyu_seika_hokoku_gaiyo/）を加工して作

線維芽細胞増殖因子に対するモノクローナル抗体の開発とその臨床応用

金沢大学がん研究所研究課題/領域番号:01770449, 研究期間(年度):1989出典：研究課題「線維芽細胞増殖因子に対するモノクローナル抗体の開発とその臨床応用」課題番号01770449（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-01770449/）を加工して作

肝細胞癌の前癌病変におけるAFP遺伝子および各種癌遺伝子の発現とその意義

金沢大学がん研究所研究課題/領域番号:03857094, 研究期間(年度):1991出典：研究課題「肝細胞癌の前癌病変におけるAFP遺伝子および各種癌遺伝子の発現とその意義」課題番号03857094（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-03857094/）を加工して作

Effect of gemcitabine on the expression of apoptosis-related genes in human pancreatic cancer cells

金沢大学がん研究所分子標的がん医療研究開発センターAim: To investigate the expression of genes involved in the gemcitabine-induced cytotoxicity in human pancreatic cancer cells. Methods: A human pancreatic cancer cell line, PANC-1, was cultured. 1 × 104 PANC-1 cells were plated in 96-well microtiter plates. After being incubated for 24 h, gemcitabine was added to the medium at concentrations ranging 2.5 -1 000 mg/L. The AlamarBlue dye method was used for cell growth analysis. DNA fragmentation was quantitatively assayed using a DNA fragmentation enzyme-linked immunosorbent assay (ELISA) kit. PAP and TP53INP1 mRNA expression was determined using the reverse transcription-polymerase chain reaction with semi-quantitative analysis. The expression of GSK-3β and phospho-GSK-3β proteins was examined with Western blot analysis. Results: The IC50 for the drug after a 48-h exposure to gemcitabine was 16 mg/L. The growth of PANC-1 cells was inhibited by gemcitabine in a concentration-dependent manner (P< 0.0001) and the cell growth was also inhibited throughout the time course (P<0.0001). The DNA fragmentation rate in the gemcitabine-treated group at 48 h was 44.7 %, whereas it was 25.3 % in the untreated group. The PAP mRNA expression was decreased after being treated with gemcitabine, whereas the TP53INP1 mRNA was increased by the gemcitabine treatment. Western blot analysis showed that phospho-GSK-3βser9 was induced by the gemcitabine treatment. Conclusion: Gemcitabine suppresses PANC-1 cell proliferation and induces apoptosis. Apoptosis is considered to be associated with the inhibition of PAP and GSK-3β, and the activation of TP53INP1 and pospho-GSK-3βser9. © 2006 The WJG Press. All rights reserved

A Kampo Medicine, Boi-ogi-to, Inhibits Obesity in Ovariectomized Rats

In women facing menopause, end of menstrual activity is accompanied by lower levels of estrogen and gradual weight gain. Postmenopausal weight gain sounds an alarm for women's health and may lead to hyperlipidemia, a lipid increase and glucose intolerance. These phenomena are connected to lifestyle-related diseases such as hypertension, type II diabetes mellitus, arteriosclerosis and metabolic syndrome, making it essential to prevent weight gain in women. A Kampo medicine, Boi-ogi-to, is traditionally used to treat obese conditions, but the mechanism has not yet been investigated. In this experiment, we tested the antiobesity properties of Boi-ogi-to in ovariectomized rats by measuring changes of serum cytokine levels and adipocytokines in fat cells. After treatment with this extract for 6 weeks (20-week-old rats), we found that there was a significant weight decrease in rats treated with Boi-ogi-to as compared with that in the control group. Serum tumor necrosis factor (TNF)-α levels increased significantly in a dose-dependent manner. Gene expression of adipose tissue in uterus also dose dependently showed a significant increase of TNF-α levels, suggesting that secretion of TNF-α by fat cells might play a role in the ability of Boi-ogi-to to inhibit weight gain. While peroxisome proliferators-activated receptor-γ and adiponectin levels did not show a significant difference as compared with those in the control, levels of mRNA expression showed a tendency to increase dose dependently. Resistin did not show any significant change. These results suggest that Boi-ogi-to might be useful for the prevention of obesity that occurs in women with reduction of estrogen