Cinnamaldehyde inhibits pancreatic cancer cell proliferation and sternness via modulation of CD44s/STAT3 signaling pathway / 中国药理学通报

Aim To explore the effects of cinnamaldehyde on the proliferation and stemness of pancreatic cancer PANC-1 cells, and its possible mechanism of action.Methods CCK8 assay was used to detect the effect of cinnamaldehyde treatment on cell viability at different concentrations(0, 5, 15, 20, 30, 50, 70, 100, 150 μmol·L-1)and different intervention time(24, 48, 72 h).CFSE proliferation assay was used to detect the inhibitory effects of cinnamaldehyde on PANC-1 cells.Colony formation assay was employed to determine the colony-forming ability of PANC-1 cells after cinnamaldehyde treatment.The sphere formation assay was employed to detect the effects of cinnamaldehyde on sphere-forming ability in PANC-1 cells.Western blot analysis and qRT-PCR analysis were applied to determine the expression levels of Nanog, Sox-2 and Oct-4.Flow cytometry was used to detect the percentage of CD44+CD24+ cells and ALDH+ cells in cinnamaldehyde treated and untreated PANC-1 cells.Western blot analysis was used to detect the effects of cinnamaldehyde on the expression of CD44s, p-STAT3 and STAT3 in PANC-1 cells.Results Cinnamaldehyde suppressed cell viability in a dose- and time-dependent manner, and inhibited tumor-cell proliferation and colony forming ability significantly in a dose-dependent manner in PANC-1 cells.Sphere-forming assay showed that cinnamaldehyde could significantly inhibit sphere-forming ability in suspension culture of PANC-1 cells.The mRNA and protein expression levels of three stemness-related genes were down-regulated after cinnamaldehyde treatment.In addition, cinnamaldehyde treatment significantly decreased the proportion of CD44+CD24+ cells and ALDH+ cells.Western blotting showed that cinnamaldehyde inhibited the expression of CD44s and p-STAT3, while it had no effect on the expression of STAT3.With the addition of STAT3 activator(Colivelin TFA), the inhibition of cinnamaldehyde on proliferation and tumor-cell stemness in PANC-1 cells was partially rescued.Conclusions Cinnamaldehyde significantly inhibits the proliferation and tumor-cell stemness of pancreatic cancer PANC-1 cells, and the mechanism could be related to the modulation of CD44s/STAT3 signaling pathway.

miR-32-5p inhibits the malignant biological behaviors and the growth of nude mice xenograft of pancreatic cancer PANC-1 cells by targeted down-regulation of EZH2 expression / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究miR-32-5p通过靶向zeste基因增强子人类同源物2（EZH2）对胰腺癌PANC-1细胞恶性生物学行为的影响及其分子机制。方法：利用GEPIA数据库分析胰腺癌组织中EZH2的表达水平及其与患者的预后生存期的关系，并分析miR-32-5p表达与患者临床病理因素的关系。qPCR法检测正常胰腺HPDE6-C7细胞和胰腺癌PANC-1、AsPC-1、SW1990细胞中miR-32-5p和EZH2 mRNA的表达，通过LipofectamineTM 2000将miR-NC、miR-32-5p mimic、miR-32-5p inhibitor、pcDNA-NC、pcDNA EZH2质粒分别转染PANC-1细胞，其分为对照组（不转染）、 miR-NC组（转染miR-NC）、 miR-32-5p mimic组（转染miR-32-5p mimic）、Anti-miR-32-5p组（转染miR-32-5p inhibitor）、miR-NC+pcDNA-NC组（转染miR-NC+pcDNA-NC）、miR-NC+pcDNA EZH2组（转染miR-NC+pcDNA EZH2）、miR-32-5p mimic+pcDNA-NC组（转染miR-32-5p mimic+pcDNA-NC）、miR-32-5p mimic+pcDNA EZH2组（转染miR-32-5p mimic+pcDNA EZH2）。双荧光素酶报告基因实验验证miR-32-5p与EZH2的靶向关系；MTT法及克隆形成实验检测各组细胞的增殖能力，划痕愈合实验检测各组细胞的迁移能力；Transwell小室实验检测各组细胞的侵袭能力，WB法检测各组细胞EZH2、E-cadherin、N-cadherin的表达；裸鼠成瘤实验检测转染后各组PANC-1细胞移植瘤的生长情况，免疫组化染色法观察移植瘤组织中Ki67和MMP-2的表达。结果：GEPIA数据库显示胰腺癌组织中EZH2的表达水平高于癌旁组织，患者预后生存期与EZH2的表达水平呈负相关（均P<0.05）；miR-32-5p表达水平与胰腺癌神经浸润、肿瘤分化程度、TNM分期、淋巴结转移有明显的关联（均P<0.05）；与HPDE6-C7细胞相比，PANC-1、AsPC-1、SW1990细胞中miR-32-5p呈高表达、EZH2 mRNA呈低表达、miR-32-5p表达水平与EZH2表达水平呈负相关（均P<0.05）。miR-32-5p靶向EZH2且抑制其表达（均P<0.05）；过表达miR-32-5p能够下调Ki67、MMP-2、N-cadherin表达水平、上调E-cadherin表达水平，抑制PANC-1细胞的增殖、迁移和侵袭能力，抑制移植瘤质量增加（均P<0.05）。结论：miR-32-5p能够靶向调控EZH2从而影响胰腺癌PANC-1细胞的增殖、迁移、侵袭、EMT进程及裸鼠体内移植瘤的生长。

CYP2J2-produced epoxyeicosatrienoic acids contribute to the ferroptosis resistance of pancreatic ductal adenocarcinoma in a PPARγ-dependent manner / 中南大学学报(医学版)

Objective: Pancreatic ductal adenocarcinoma (PDAC) is one of the most malignant digestive tract tumors with a poor prognosis and high recurrence rate. Recently, ferroptosis resistance has been found in PDAC. However, the underlying mechanism of ferroptosis resistance has not been fully elucidated. Cytochrome P4502J2 (CYP2J2) is the main enzyme which mediates arachidonic acid to produce epoxyeicosatrienoic acids (EETs) in human tissues. It has been reported that EETs involve in the development of cancer, while the roles of EETs in PDAC and ferroptosis remain unclear. This study aims to explore the effect of CYP2J2/EETs on ferroptosis of human pancreatic ductal adenocarcinoma cells PANC-1 cells and the underlying mechanisms.Methods: The tumor tissues and para-carcinoma tissues of 9 patients with PDAC were collected and the expression of CYP2J2 was detected with real-time PCR and Western blotting. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of 8,9-dihydroxyeicosatrienoic acid (8,9-DHET), and the degradation product of 8,9-epoxyeicosa-trienoic acid (8, 9-EET). PANC-1 cells were used in this study. The ferroptosis inducer erastin was used to induce ferroptosis. The intracellular long-chain acyl-CoA synthetase 4 (ACSL4) protein level, lactate dehydrogenase (LDH) activity, malondialdehyde (MDA) content, Fe2+concentration, and cell survival were detected. The 8, 9-EET was pretreated to observe its effect on erastin-induced ferroptosis in PANC-1 cells. Lentivirus was used to construct a CYP2J2 knockdown cell line to observe its effect on the ferroptosis of PANC-1 cells induced by erastin. A peroxisome proliferation-activated receptor γ (PPARγ) blocker was used to observe the effect of 8, 9-EET on erastin-induced glutathione peroxidase 4 (GPX4) and MDA content in PANC-1 cells.Results: High expression of CYP2J2 was found in PDAC, accompanied by an increased level of 8, 9-DHET. The 8, 9-EET pretreatment significantly attenuated the PANC-1 cell death induced by erastin. The 8, 9-EET reduced the Fe2+ concentration, LDH activity and MDA content, and ACSL4 protein expression in erastin-treated PANC-1 cells. The 8,9-EET also restored the ferroportin (FPN) and ferroptosis suppressor protein 1 (FSP1) mRNA expressions in erastin-treated PANC-1 cells. But CYP2J2 knockdown exacerbated the erastin-induced ferroptosis in PANC-1 cells. Besides, CYP2J2 knockdown furtherly down-regulated the gene expression of FPN and FSP1. The 8, 9-EET increased the expression of GPX4 in the erastin-treated PANC-1 cells, which was eliminated by a PPARγ blocker GW9662. And GW9662 abolished the anti-ferroptosis effects of 8,9-EET. Conclusion: CYP2J2/EETs are highly expressed in PDAC tissues. EETs inhibit the ferroptosis via up-regulation of GPX4 in a PPARγ-dependent manner, which contributes to the ferroptosis resistance of PDAC.

Role of PANC-3 score to predict severe acute pancreatitis

Background: Acute pancreatitis is the most terrible of all the calamities that occur in connection with the abdominal viscera. Prediction of severity is an essential step in the management of acute pancreatitis. 50% of mortality can be reduced to 8% by its early recognition. PANC-3 score is widely available test that can be performed quickly, easy to measure with high accuracy in predicting acute pancreatitis.Methods: This cross-sectional study was conducted in the department of general surgery, VMMC and Safdarjung Hospital over 50 patients admitted with acute pancreatitis. After making the clinical diagnosis, PANC -3 score, modified ATLANTA score, APACHE II were done. CRP and CTSI (computed tomography sensitivity index) were calculated and correlated.Results: Mean age was 44.74 years and most common cause was biliary tract pathology. Mortality observed in 5 patients, 11 patients had severe disease. Sensitivity of PANC- 3 was 81.82%, specificity -92.31% with 75% PPV and 94.7% NPV.Conclusions: PANC-3 can be used to predict the severity of pancreatitis as efficiently as Modified ATLANTA classification/APACHE II. It uses only three criteria which are easily done, and available in the basic health care setup. Its interpretation does not need expertise and can be applied at the time of admission which is an advantage when compared to classical scoring systems.

Essential oil composition and cytotoxic activity in twospecies ofthe plant genus Vismia (Hypericaceae) from the Venezuelan Andes / Composición de aceites esenciales y actividad citotóxica en dos especies de plantas del género Vismia (Hypericaceae) de los Andes venezolanos

Introduction: The Vismia genus belongs to the Hypericaceae family and comprises around 57 species of which 17 have been located in Venezuela. Previous investigations have been carried out in extracts as well as pure isolated compounds, revealing antimicrobial, antioxidant and anti-HIV, among other, biological activities. Objective: This investigation aims to determine the cytotoxic activity of essential oils from leaves of Vismia baccifera Triana & Planch (VBJ and VBV) and Vismia macrophylla Kunth (VM) collected in three different locations of the Venezuelan Andean region. Methods: Essential oils obtained by hydrodistillation were analyzed using gas chromatography-mass spectrometry (GC-MS) and their cytotoxic activity was analyzed following the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Human tumor cell lines from SKBr3, MCF-7 and PANC-1, two breast carcinomas and one pancreatic adenocarcinoma of ductal type, were tested with the oil samples and human dermis fibroblasts were used as non-tumor cells. Results: β-caryophyllene and trans-caryophyllene were present as major components in VBJ and VBV, respectively, while γ-bisabolene was the main component in the VM sample. Anticancer activity was observed on V. baccifera essential oil against SKBr3, MCF-7 and PANC-1. The selectivity index showed that VBV is highly selective against the SKBr3 cell line and has no activity against non-tumor cells. Conclusions: These results are considered a contribution to natural products research and may provide supportive data for future studies on cancer.

Introducción: El género Vismia pertenece a la familia Hypericaceae y comprende alrededor de 57 especies de las cuales 17 han sido ubicadas en Venezuela. Se han realizado investigaciones previas tanto en extractos como en compuestos puros aislados revelando actividad antimicrobiana, antioxidante y anti-VIH, entre otras actividades biológicas. Objetivo: El propósito de esta investigación es determinar la actividad citotóxica de los aceites esenciales de las hojas de Vismia baccifera Triana & Planch (VBJ y VBV) y Vismia macrophylla Kunth (VM) recolectadas en tres localidades de la región andina venezolana. Métodos: Aceites esenciales obtenidos por hidrodestilación fueron analizados por cromatografía de gases-espectrometría de masas y su actividad citotóxica fue analizada siguiendo el método MTT (bromuro de 3-[4,5-dimetiltiazol-2-il]-2,5-difeniltetrazolio). Los aceites esenciales fueron ensayados frente a células tumorales humanas SKBr3, MCF-7 y PANC-1, dos carcinomas de mama y un adenocarcinoma pancreático del tipo ductal, usando cultivos primarios de fibroblastos de dermis humana como células no tumorales. Resultados: β-cariofileno y trans-cariofileno estuvieron presentes como compuestos mayoritarios en VBJ y VBV, respectivamente, mientras que γ-bisaboleno fue el componente principal en la muestra VM. El aceite esencial de V. baccifera mostró actividad anticancerígena frente a SKBr3, MCF-7 y PANC-1. El índice de selectividad reveló que VBV es altamente selectivo frente a la línea celular SKBr3 y no presenta actividad contra las células no tumorales. Conclusiones: Estos resultados se consideran una contribución a la investigación de los productos naturales y los datos pueden ser de utilidad en futuras investigaciones sobre el cáncer.

Sustentabilidade: utilização de vegetais na forma integral ou de partes alimentícias não convencionais para elaboração de farinhas / Sustainability: use of vegetables in their entirety or non-coventional food parts for flours preparation

Introdução: O desperdício é um grande problema enfrentado pelo Brasil, em função dos hábitos alimentares, sendo o aproveitamento de cascas de frutas, folhas e talos de hortaliças ainda baixo. Sabe-se, no entanto, que a utilização dessas partes tradicionalmente não aproveitadas pode contribuir para a produção de alimentos mais saudáveis, nutritivos e com menor impacto negativo para o meio ambiente. Objetivo: Aproveitar os vegetais na sua forma integral ou de suas partes alimentícias não convencionais, e caracterizar as farinhas obtidas, com vistas à alimentação humana. Métodos: As farinhas foram obtidas de banana verdes (casca e polpa), casca de tangerina, casca e polpa de berinjela e bagaço de uva oriundo da produção de suco de uva, que foram secas em estufa ventilada a 40°C por 48 horas. Foram realizadas análises físico-químicas e microbiológicas. Resultados: As farinhas apresentaram alto teor de fibras (17,7 a 48,94 g/100g) e compostos fenólicos (778,81 a 2708,11 mg EAG/100g), e baixo teor de lipídeos (0,70 a 3,40 g/100g). Para a quantidade de proteínas, a farinha de berinjela foi a que mais se destacou (13,81 g/100g), podendo ser uma alternativa aos indivíduos que não consomem produtos de origem animal. Conclusões: Todas as farinhas são seguras do ponto de vista microbiológico, de acordo com a legislação vigente, e portanto, podem ser utilizadas na alimentação humana para aumentar o aporte de nutrientes e diminuir os impactos da poluição, contribuindo para a manutenção do meio ambiente e gerando renda. (AU)

Introduction: Waste is a great problem faced by Brazil, due to dietary habits, with a low use of fruit peels, leaves and stems of vegetables. However, it is known that the use of these parts traditionally not used can contribute to the production of healthier and more nutritious food with less negative impact on the environment. Objective: Take advantage of vegetables in their entirety or from their non-conventional food parts, and to characterize the flours obtained, for human consumption. Methods: Flours were obtained from green bananas (peel and pulp), tangerine peel, eggplant peel and pulp and grape marc from grape juice production, which were dried in a ventilated oven at 40 ° C for 48 hours. Physical-chemical and microbiological analyzes were realized. Results: Flours showed a high fiber (17.7 to 48.94 g/100g) and phenolic compounds (778.81 to 2708.11 mg EAG/100g) content, and low lipid content (0.70 to 3.40 g/100g). For the amount of proteins, eggplant flour was the one that stood out the most (13.81g/100g), it can be an alternative to individuals who do not consume animal products. Conclusions: All flours are safe in a microbiological point of view, according to current legislation, and therefore they can be used in food to increase the supply of nutrients and reduce the impacts of pollution, contributing to the environment maintenance and income generating. (AU)

Plantas alimentícias não convencionais no Brasil: o que a Nutrição sabe sobre este tema? / Unconventional food plants in Brazil: what does Nutrition know about this topic

As plantas alimentícias não convencionais, conhecidas pela sigla PANC, são hortaliças, frutas, flores ou ervas que crescem espontaneamente na natureza, mas que por serem desconhecidas para a maioria das pessoas, acabam sendo confundidas com plantas daninhas. Este estudo objetivou realizar um levantamento das pesquisas sobre PANC nos anais das últimas seis edições do Congresso Brasileiro de Nutrição (CONBRAN) e nos anais das últimas sete edições do Congresso da Sociedade Brasileira de Nutrição (SBAN), indicando o perfil dessas pesquisas no Brasil no campo da Nutrição. Trata-se de pesquisa qualitativa, descritiva e exploratória. Foram identificados 25 trabalhos sobre o tema no CONBRAN e dez no Congresso SBAN, indicando que as PANC são tema ainda pouco explorado e abordado com pouca frequência nas pesquisas em Nutrição no Brasil. Destaca-se o potencial emergente de pesquisa que as PANC podem representar no campo da Alimentação em suas mais diversas vertentes: culinária, gastronomia, segurança alimentar, ciência de alimentos, entre outras. Esta pesquisa contribui para a ampliação do conhecimento e direcionamento dos estudos nesta área e, assim, poderá sinalizar uma agenda para estudos futuros. (AU)

Unconventional food plants, known by the acronym UFP (PANC, in Portuguese), are vegetables, fruits, flowers or herbs that grow spontaneously in nature, but because they are unknown to most people, they end up being confused with weeds. This study aimed to carry out a survey of the research on non-conventional food plants in the records of the last six editions of the Brazilian Congress of Nutrition (CONBRAN) and of the last seven editions of the Congress of the Brazilian Society of Nutrition (SBAN), indicating the profile of these studies in Brazil in the field of Nutrition. It is a qualitative, descriptive and exploratory research. Twenty-five studies on the subject at the CONBRAN Congress and ten at the SBAN Congress were identified, indicating that UFP still are little explored subject and rarely addressed in Nutrition research in Brazil. We highlight the emerging research potential that UFP can represent in the field of Food in its most diverse aspects: cooking, gastronomy, food safety, food science, among others. This research contributes to the expansion of knowledge and direction of studies in this area and it may also signal an agenda for future studies. (AU)

Effects of Gentiopicroside on the Apoptosis of Human Pancreatic Cancer Cells PANC- 1 and IL- 6/JAK2/STAT3 Signaling Pathway / 中国药房

OBJECTIVE：To study the effects of gentiopicroside on the apo ptosis o f human pancreatic cancer cells PANC- 1，and to explore its mechanism from the perspective of IL- 6/JAK2/STAT3 signaling pathway. METHODS ：Using PANC- 1 cells as model ， the proliferation inhibition rate of cells was tested by MTT assay after treated with 0（negative contro ），2，4，8，16，32，64，128 mg/L gentiopicroside for 72 h and IC 50 were calculated. The cells were divided into negative control group ，gemcitabine group （positive control，4 mg/L）and gentiopicroside low-concentration ，medium-concentration and high-concentration groups （15，30，60 mg/L）. After cultured for 1，3，5，7 d，Trypan blue exclusion staining was used to count the survival cell ，and the growth of cells was investigated. After cultured for 72 h，colony formation assay was used to observe colony formation rate of cells ；the apoptotic rate of cells was detected by Hoechst 33258 staining；the mRNA and protein expressions of IL- 6，JAK2，STAT3 in cells were detected by RT-PCR and Western blotting assay. RESULTS ：4-28 mg/L gentiopicroside could inhibit the proliferation of cells （P＜0.05 or P＜ 0.01），in concentration dependent trend ；IC50 was 9.54 mg/L. Compared with negative control group ，survival cell count （cultured from 3，5，7 d），mRNA and protein expressions of IL- 6，JAK2 and STAT 3 in cells were decreased significantly in gemcitabine group ， gentiopicroside medium-concentration and high-concentration groups （P＜0.05 or P＜0.01），while the apoptotic rate was increased significantly （P＜0.01）. The colony formation rate of cellswere decreased significantly in gemcitabine group and gentiopicroside high-concentration group （P＜0.01）. mail：hb.gz@163.com Compared with gemcitabine group ，there was no statistical significance in above indexes of gentiopicroside high- 6716008。 concentration group （P＞0.05）. CONC LUSIONS：30，60 mg/L gentiopicroside could inhibit the proliferation and induce apoptosis of PANC- 1 cells，and 60 mg/L gentiopicroside is similar to gemcitabine in the effects. Its mechanism may be related to inhibiting the activation of IL- 6/JAK2/STAT3 signaling pathway.

Características físico-químicas dos tubérculos da Tropaeolum pentaphyllum Lam / Physico-chemical characteristics of tubers of Tropaeolum pentaphyllum Lam

A Tropaeolum pentaphyllum Lam é uma Planta Alimentícia Não Convencional (PANC) nativa do Brasil, que está em crescente estudo. O objetivo deste trabalho foi avaliar a composição físico-química, o pH e cor da polpa da batata. Observou-se que a umidade (79,32 %), proteína (2,33%) e cinzas (0,93%), apresentaram-se semelhantes à T. pentaphyllum Lam avaliadas em outros estudos. Já os teores de carboidratos (13,02%), valor energético (82,19%) e lipídeos (3,82%) apresentaram-se superiores e de fibras (1,78%) inferiores a vegetais da mesma espécie. Além disso, a planta mostrou-se ácida, luminosa e com coloração tendendo ao vermelho e ao amarelo. Assim, percebe-se que a batata pode ser considerada um complemento alimentar pela sua composição nutricional e que é necessário a realização de mais estudos, a fim de aprimorar o conhecimento sobre a mesma.

Análise de correspondência aplicada para a identificação de plantas benéficas da região de Salinas-MG / Correspondence analysis applied for the identification of beneficial plants of the Salinas-MG region

Plantas nativas são utilizadas na alimentação humana e na cura de enfermidades. No que diz respeito às plantas medicinais, o emprego das substâncias nelas contidas, advém do conhecimento empírico. Já as PANC’s são conhecidas pela infinidade de benefícios à saúde, no entanto, há uma variedade de compostos desconhecidos por parte da comunidade científica. Diante do mencionado, objetivou-se coletar informações acerca das plantas com propriedades benéficas e, posteriormente dissipar este conhecimento. Foram feitas visitas in loco para obtenção de dados, e estes tratados pela técnica estatística de análise de correspondência. Foi feito, ainda, uma pesquisa na literatura para a comprovação dos dados encontrados na análise, e, por fim foram criadas cartilhas contendo as informações em relação a utilização das plantas endêmicas da região.

Betulinic acid enhances gefitinib-sensitivity of pancreatic cancer cells via inhibition of STAT3 activation / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effects and the underlying mechanisms of betulinic acid (BEA) on sensitizing pancreatic cancer cell lines Panc-1 and Miapaca-2 to gefitinib. Methods: After the cell culture was completed, Panc-1 and Miapaca-2 cells were randomly divided into 4 groups: control group (without treatment), BEAgroup, gefitinib group and BEAcombined with gefitinib group, respectively.The sensitization effect of BEAon gefitinib-insensitive pancreatic cancer cells was detected by MTS assay. The treatment effects of combined treatment of gefitinib and BEA against Panc-1 and Miapaca-2 cells were evaluated by colony formation assay. Flow cytometry was used to examine the effect of BEAon apoptosis of Panc-1 cells while WB was applied to determine the effect of BEAonapoptosis-related proteins. Surface plasmon resonance (SPR) experiment was used to detect the direct combination between signal transducer and activator of transcription 3(STAT3) and BEA; Molecular docking and molecular dynamics simulation experiments were adopted topredict the combining mode between STAT3 and BEA. Results: BEA synergistically enhanced the gefitinib-sensitivity of pancreatic cancer Panc-1 and Miapaca-2 cells (P<0.05), and IC50 of gefitinib on two cells were reduced by over 50%. Compared with single treatment, the combined treatment of BEA and gefitinib promoted the apoptosis and up-regulated the expressions of apoptosis-relatedproteins (cleaved-PARP and Bax), but reduced the apoptosis-inhibitory protein Bcl-2 (all P<0.05 or P<0.01). Moreover, the inhibitory effect of BEA on STAT3 activation in Panc-1 cells was in a dose-dependent mannar (P<0.01). BEA stabilizes its binding to STAT3 by forming hydrogen bonds with Lys-591 and Ser-613 of STAT3; in the meanwhile, BEA stabilized inthebinding site of STAT3, there by blocking STAT3 dimerization to enhance the drug sensitivity. Conclusion: Combined use of BEA and gefitinib could significantly inhibit the proliferation and induce apoptosis of Panc-1 and Miapaca-2 cells, which might be mediated by the inhibition of BEA on STST3.

Anti-pancreatic fibrosis of oxymatrine by promoting expression of transcription factors Gli2 in LTC-14 and PANC-1 pancreatic stellate cells / 中草药

Objective: To investigate the effect of oxymatrine (OM) on the expression of Gli2 in LTC-14 cells and PANC-1 cells induced by TGF-β1. Methods: LTC-14 cells and PANC-1 cells were divided into four groups:control group, TGF-β1 group, TGF-β1 + OM group, and OM group. The protein was extracted after 12 h, and the expressions of related proteins were detected by Western blot. Results: Compared with control group, the expression of Gli2 in LTC-14 cells was significantly decreased induced by TGF-β1, while the expressions of α-SMA, FN, and CoL-I were significantly increased. The expressions of Gli2, FN, CoL-I, and α-SMA were increased significantly induced by TGF-β1when compared with those of control group in PANC-1 cells, and the expression of Gli2 was inhibited by OM pretreatment in TGF-β1 + OM group compared with TGF-β1 group. OM pretreatment can increase the expression of Gli2 before stimulating with TGF-β1 in TGF-β1 + OM group in comparison with TGF-β1 group. Conclusion: OM plays an protected role in pancreatic fibrosis through promoting the expression of Gli2 in LTC-14 cells.

miR-429 promotes capecitabine-resistance in pancreatic cancer PANC-1 cells by down-regulating PTEN and activating PI3K/AKT signaling pathway / 中国肿瘤生物治疗杂志

@# Objective: To explore the mechanism of miR-429 targeting PTEN to affect capecitabine-resistance in pancreatic cancer PANC-1 cells though the PI3K/AKT signaling pathway. Methods: Capecitabine-resistant pancreatic cancer cell line PANC-1/CAP was constructed, and the expression of miR-429 and PTEN were detected by quantitative Real-time polymerase chain reaction (qRT-PCR) and Western blotting. The effect of miR-429 knock-down on cell proliferation viability, apoptosis and capecitabine-resistance was measured by colony formation assay, CCK-8 assay andAnnexin V-FITC/PI double staining flow cytometry assay, respectively. Subsequently, dual luciferase reporter assay verified that PTEN was a target gene of miR-429. Furthermore, the effect of miR-429 on PTEN-PI3K/ AKT signaling pathway was measured by Western blotting. Results: miR-429 was found to be up-regulated in PANC-1 cells and PANC-1/CAP cells compared with the non-malignant pancreatic ductal cell line (HPDE6-C7) (P<0.05 or P<0.01). Moreover, silencing of miR-429 significantly decreased cell proliferation viability, capecitabine-resistance and enhanced apoptosis of PANC-1/CAP cells; additionally, dual luciferase reporter assay confirmed that PTEN was a target of miR-429 (P<0.05 or P<0.01). Suppression of miR-429 up-regulated PTEN and blocked the PI3K/AKT signaling pathway to decrease cell proliferation viability and further reduce the capecitabine-resistance of PANC-1/CAP cells (P<0.05 or P<0.01). Conclusion: miR-429/PTEN-PI3K/AKT signaling pathway plays a certain role in regulating the capecitabine-resistance of pancreatic cancer, and inhibition of miR-429 expression may reverse the resistance of PANC-1/CAPto capecitabine.

Effect and mechanism of cetuximab on cell proliferation of SW1990 and PANC-1 cells in pancreatic carcinoma / 中国生化药物杂志

Objective To investigate the effect of cetuximab on the proliferation of pancreatic carcinoma cell lines SW1990 and PANC-1.Methods 100.0 μg/mL cetuximab added DMEM medium(cetuximab group), simple DEME medium as the control group, two groups of the pancreatic carcinoma cell lines SW1990 and PANC-1 were cultured, the SW1990 and PANC-1 were detected by MTT after cultured 12, 24, 48 h, the expression of epidermal growth factor receptor gene(EGFR-mRNA)and protein in two groups were detected by RT-PCR and Western-blot after 24 h;the cell cycle was detected by flow cytometry after 24 h, the activity of Wnt/ beta-catenin pathway was detected by double luciferase assay after 24 h.Results The OD of the cetuximab group of the SW1990 and PANC-1 cell lines proliferation were lower than the control group after 12, 24, 48 h (P<0.05);the OD of the cetuximab group of the SW1990 and PANC-1 cell lines proliferation in 24, 48 h was significantly decreased than 12 h (P<0.05);the proportion of G1 phase cells of the SW1990 and PANC-1 of the cetuximab group increased significantly than the control group (P<0.05), the proportion of S and G2 phase cells decreased significantly than the control group (P<0.05), the expression of the EGFR-mRNA and protein of the cetuximab group were lower than the control group (P<0.05),the TOP/FOP of the Wnt/beta-catenin pathways' activity of the cetuximab group was lower than that of the control group (P<0.05).Conclusion The proliferation of pancreatic carcinoma cell lines SW1990 and PANC-1 was significantly inhibited by cetuximab, which was mainly achieved by decreasing the activity of Wnt/beta-catenin pathway and the expression of EGFR-mRNA and protein.

Effects of Ginsenoside CK Combined with 5-Fluorouracil on the Proliferation,Apoptosis and Epithelial Mesenchymal Transition of Human Pancreatic Cancer PANC-1 Cells / 中国药房

OBJECTIVE:To study the effects of ginsenoside CK combined with 5-fluorouracil (5-FU) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of human pancreatic cancer PANC-1 cells. METHODS:PANC-1 cells of logarithmic growth phase were randomly divided into blank control group,ginsenoside CK group (30 mg/L),5-FU group (25 mg/L)and combination group(ginsenoside CK 30 mg/L+5-FU 25 mg/L). MTT method was used to detect the cell proliferation in-hibition rate in each group after 24,48,72 h;flow cytometry was used to detect the cell apoptosis rate after 48 h;enzyme-linked immunosorbent assay was used to detect the fibronectin expression in cells after 24,48,72,96 h;and Western blot was used to detect the expressions of vimentin,N-cadherin,E-cadherin,protein kinase(Akt)and phosphorylated Akt(p-Akt)protein in cells after 48 h. RESULTS:Compared with blank control group,the cell proliferation inhibition rate,early and late apoptotic rates,pro-tein expression level of E-cadherin in ginsenoside CK group,5-FU group and combination group were obviously increased (P<0.05),while the protein expression levels of fibronectin,vimentin,N-cadherin,and p-Akt/Akt levels were obviously decreased (P<0.05);the effects of above-mentioned indexes in combination group were superior to ginsenoside CK group and 5-FU group (P<0.05). CONCLUSIONS:Both ginsenoside CK and 5-FU can inhibit the proliferation of PANC-1 cells,induce their apoptosis and inhibit EMT,which may be associated with inhibiting phosphatidylinositol 3-kinase/Akt pathway. In addition,the combination of ginsenoside CK and 5-FU can produce a better effect.

Effects of oxymatrine on expression of related molecules in Smad3/Gli1 signaling pathway in PANC-1 cells induced by TGF-β1 / 中草药

Objective: To investigate the effect of oxymatrine (OM) on expression of related molecules in Smad3, Gli1 signaling pathway in PANC-1 cells induced by TGF-β1. Methods: TGF-β1-induced PANC-1 cells were used to establish the pancreatic fibrosis model in vitro, and observe the effects of OM pretreatment on the related molecular expression of Smad3/Gli1. Gli1 and Smad3 RNA interference plasmids were transfected into PANC-1 cells. The protein expression levels of Smad3, Gli1 and α-SMA were measured by Western blotting. The levels of fibronectin (FN) and type I collagen (CoL-I) in the supernatant of cell culture were detected by ELISA. Results: Compared with the control group, the protein expressions of Smad3, Gli1 and α-SMA increased significantly in PANC-1 cells after treated with TGF-β1. The expressions of Gli1, α-SMA, FN, and CoL-I in PANC-1 cells decreased significantly after Gli1 RNA interference plasmid transfection compared with TGF-β1 induced group. The expression of Smad3, Gli1, α-SMA, FN, and CoL-I also decreased significantly in PANC-1 cells after Smad3 RNA interference plasmid transfection compared with TGF-β1 group. Conclusion: OM could prevent pancreatic fibrosis by regulating TGF-β1/Smad3/Gli1 signaling pathway in PANC-1 cells.

VX-2 tissue orthotopic implantation versus panc-1 cell suspension orthotopic implantation in establishing a rabbit model of pancreatic cancer / 医学研究生学报

Objective Few reports are seen on the methods of establishing the rabbit model of pancreatic cancer .This study was to compare the effect of Panc-1 cell suspension orthotopic implantation with that of VX-2 tissue orthotopic implantation in construc-ting the rabbit model of pancreatic cancer . Methods Using the random number table method , we divided 30 healthy rabbits into a tissue suspension group ( n=15) and a cell suspension group ( n=15) , VX-2 tissue suspension employed for in-situ implanting in the former group and panc-1 cell suspension utilized in the latter .Then we evaluated the two modeling methods by B-ultrasonography , 3.0T MRI, and CT. Results In the third week after modeling , transpla-ntive metastasis of lots of tumor tissues was observed in the duode-num, colon, appendix, and peritoneal wall in 5 rabbits of the tissue suspension group , but only in the greater omentum of 3 rabbits in the cell suspension group , with high signals of MR T 2 in the posterior gastric body .One case of duodenal metastasis was seen in the cell suspension group , with slightly high signals of MR LAVA in the posterior gastric body .The model of pancreatic cancer was successfully established in all the 15 rabbits of the tissue suspension group , but only in 3 of the cell suspension group .The success rate of tumor im-planting at 3 and 4 weeks was significantly higher in the former ( 46.66%and 100%) than in the latter group ( 6.67%and 20.00%) (P<0.05). Conclusion VX-2 tissue orthotopic implantation is a more feasible and convenient method than Panc -1 cell suspension orthotopic implantation for establishing the rabbit model of pancreatic cancer .

Effect of DPF2-RNAi on proliferation, apoptosis and cell cycle of PANC-1 cells / 中国药理学通报

Aim DPF2 has been reported to be involved in pathogenesis of leukaemia and oncogenic activity.However,the role of DPF2 in oncogenesis and pathogenesis of pancreatic cancer remains unclear.Therefore,the present research aims to investigate the effects of DPF2-RNAi on proliferation,apoptosis and cell cycle regulation of a pancreatic cell line,PANC-1 cells.Methods The lentivirus-mediated DPF2-RNAi was employed to knockdown DPF2 expression in PANC-1 cells,and the role of DPF2-RNAi in proliferation,apoptosis and cell cycle regulation of the PANC-1 cells was examined through MTT assay,colony formation assay and flowcytometry assay.Results The lentivirus-mediated DPF2-RNAi middle and high doses(2 μL and 4 μL)significantly decreased the expression of DPF2 in the PANC-1 cells.DPF2-RNAi decreased cell viability and colony formation,and increased apoptosis of the PANC-1 cells.Besides,DPF2-RNAi induced the S-phase arrest and decreased G2/M phase population of the PANC-1 cells.Conclusions DPF2 may play a crucial role in proliferation,apoptosis and cell cycle regulation of PANC-1 cells.Knockdown of DPF2 through lentivirus-mediated DPF2-RNAi may provide experimental basis for finding a new method for therapy of pancreatic cancer.

Effect of Rip2 overexpression on apoptosis in human pancreatic cancer cell line Panc-1 / 中国病理生理杂志

AIM: To observe the effect of receptor-interacting protein 2 (Rip2) overexpression on human pan-creatic cancer cell line Panc-1.METHODS: pEGFP-C2 and pEGFP-Rip2 plasmids were respectively transfected into the Panc-1 cells using JetPRIME reagent.The cells were divided into control group, pEGFP-C2 group and pEGFP-Rip2 group. The apoptosis in the cells was detected 48 h after transfection by flow cytometry.Rip2 level and the expression of apoptosis-related proteins, Bax, cytoplasmic cytochrome c ( Cyt-c) and Bcl-2, were analyzed by Western blot.The activity of caspase-3 was measured by colorimetric method.RESULTS: Rip2 protein expression significantly increased in the cells transfected with control and pEGFP-C2 plasmids.The apoptotic rate in pEGFP-Rip2 group was higher than that in control group and pEGFP-C2 group, whereas no significant difference of apoptotic rate was observed between control group and pEGFP-C2 group.The protein expression of Bax and cytoplasmic Cyt-c was remarkably increased and the protein expression of Bcl-2 was obviously decreased in pEGFP-Rip2 group as compared with control group and pEGFP-C2 group.The activity of caspase-3 in pEGFP-Rip2 group was obviously increased as compared with control group and pEGFP-C2 group.CON-CLUSION: Overexpression of Rip2 is able to induce apoptosis in the Panc-1 cells, and the mechanism may be related to the up-regulation of Bax and cytoplasmic Cyt-c protein expression, down-regulation of Bcl-2 protein expression and en-hancement of caspase-3 activity, thus activating intrinsic apoptotic pathway.

Effects ofβ-Elemene on Apoptosis of Human Pancreatic Cancer Panc-1 Cells / 中国中医药信息杂志

Objective To investigate the effects of β-Elemene on the apoptosis of human pancreatic cancer Panc-1 cells; To discuss its mechanism of action. Methods β-Elemene (10, 20, 40, 80, 160 μg/mL) were incubated to the Panc-1 cells in vitro cultured for 24 h, 48 h and 72h, and trypan blue refusal method was used to detect cell inhibition rate;Apoptosis rate was measured by TUNEL; Hoechst33258 fluorescent staining was used to observe the changes of the nucleus. The activity of Caspase-3, 8 and 9 were detected by ELISA. Western blot was used to detect the expressions of Fas, FasL and Cyt c and AIF. Results The activity of Panc-1 cells was obviously inhibited time/concentration dependent inhibition (P<0.05, P<0.01), and the apoptosis rate increased after incubated with β-Elemene (P<0.01, P<0.001) after incubated with β-Elemene for 24 h, 48 h and 72 h; After giving β-Elemene 72 h, Panc-1 cells nucleus were broken obviously, and chromatin condensed and showed strong blue fluorescence, along with of apoptotic bodies; After incubated with β-Elemene for 48 h, Caspase-3, 8 and 9 activity significantly increased (P<0.05, P<0.01); protein expressions of Fas, FasL, Cyt c and AIF were significantly enhanced (P<0.05, P<0.01, P<0.001). Conclusion β-Elemene can inhibit Panc-1 cell proliferation, induce apoptosis, and the mechanism may be related to activating cell death receptor pathway and mitochondrial apoptosis pathway to play anti-tumor effects.