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Tetracycline-inducible protein expression in pancreatic cancer cells: Effects of CapG overexpression

World Journal of Gastroenterology 2011年第15期17卷 1947-1960页

作者： Sarah Tonack Sabina Patel Mehdi Jalali Taoufik Nedjadi Rosalind E Jenkins Christopher Goldring John Neoptolemos Eithne Costello Liverpool CR-UK Centre Department of Molecular and Therapeutic Cancer MedicineUniversity of Liverpool Liverpool L69 3GA United Kingdom MRC Centre forDrug Safety Science Department of Pharmacology and Therapeutics School of Biomedical Sciences University of Liverpool L69 3GA United Kingdom National Institute for Health Research pancreatic Biomedical Research Unit Liverpool L69 3GA United Kingdom

AIM: To establish stable tetracycline-inducible pancreatic cancer cell lines. METHODS: Suit-2, MiaPaca-2, and Panc-1 cells were transfected with a second generation reverse tetracycline-controlled transactivator protein (rtTA2S-M2), under the control of either a cytomegalovirus (CMV) or a chicken β-actin promoter, and the resulting clones were ***: Use of the chicken (β-actin) promoter proved superior for both the production and maintenance of doxycycline-inducible cell lines. The system proved versatile, enabling transient inducible expression of a variety of genes, including GST-P, CYP2E1, S100A6, and the actin capping protein, CapG. To determine the physiological utility of this system in pancreatic cancer cells, stable inducible CapG expressors were established. Overexpressed CapG was localised to the cytoplasm and the nuclear membrane, but was not observed in the nucleus. High CapG levels were associated with enhanced motility, but not with changes to the cell cycle, or cellular proliferation. In CapG-overexpressing cells, the levels and phosphorylation status of other actin-moduating proteins (Cofilin and Ezrin/Radixin) were not altered. However, preliminary analyses suggest that the levels of other cellular proteins, such as ornithine aminotransferase and enolase, are altered upon CapG induction. CONCLUSION: We have generated pancreatic-cancer derived cell lines in which gene expression is fully controllable.

关键词： pancreatic cancer cells Tetracycline-inducible CapG Suit-2 Panc-1 MiaPaCa-2

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In Vitro Lethal Effect of Photodynamic Therapy on Human pancreatic cancer cells and Its Major Influencing Factors

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Clinical oncology and cancer researeh 2011年第3期8卷 155-162页

作者： Zhong YU Kai-hong HUANG Wa ZHONG Li-qun YANG Qi-kui CHEN Zhao-hua ZHU Department of Gastrointestinal Medicine Zhongshan University Sun Yat-sen Memorial Hospital Guangzhou 510120 China Zhongshan University School of Chemistry and Chemical Engineering Science Guangzhou 510080 China

OBJECTIVE To investigate the in vitro lethal effect of photo- dynamic therapy （PDT） using the photosensitizer hematoporphyrin on the human pancreatic cancer cell line Panc-1, the major influencing factors and the mechanisms of treatment. METHODS Three factors--the time needed for photosensitizer and cell incubation, the photosensitizer concentration （PhoC） and the exposure dose （ExpD）--were examined with different levels of these factors. Optical density （OD） was used as a measure of CCK-8 in the experiment, and was converted to the rate of cell survival. The separate effect of each factor on the photodynamic action was studied, and the interactions were investigated. The effects of different incubation times and PhoC levels on the fluorescence intensity （FI） of the intracellular photosensitizer were determined, and the mechanisms of these factors leading to the therapeutic effects of PDT discussed. RESULTS An increase in the photosensitizer and cell incubation time, an increase of PhoC, and enhancement of the ExpD, produced a corresponding decrease in the rate of Panc-1 cell survival after PDT （P 〈 0.05）. PDT achieved its maximum lethal effects 16 h after starting the incubation, with a PhoC of 10 mg/L and an ExpD of 20 J/cm2; at these levels a synergistic interaction between PhoC and the ExpD occurred, decreasing the cell survival rate （P 〈 0.05）. Neither simple administration of photosensitizer without ExpD （0 J/cm2） or illumination in the absence of PhoC （0 mg/L） affected the rate of cell survival （P 〉 0.05）. With an increase of PhoC and lengthening of the incubation time, the FI of the intracellular photosensitizer accordingly increased （P 〈 0.05）, and attained its maximum value at a PhoC of 10 mg/L and 36 h after the incubation. With an increase of PhoC, the FI of the photosensitizer, hematoporphyrin, in the solution increased progressively at first and then decreased （fluorescence quenching）. CONCLUSION PDT with the photosensitizer hematoporphyrin has clear l

关键词： pancreatic cancer cells photodynamic therapy photosensitizer fluorescence quenching

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Preparation and in vitro antitumor activities of triptolide and cyclopamine loaded polymeric micelles

Preparation and in vitro antitumor activities of triptolide ...

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2018年第十二届中国药物制剂大会

作者：刘德俊 Di Zhang Yixue Pei Yongjie Liu Lingyun Xu School of Biology and Pharmaceutical Engineering Wuhan Polytechnic University

Objective: In order to overcome the poor solubility of triptolide(TP) and cyclopamine(CPA), enhance their anticancer effects, and decrease their side effects, triptolide and cyclopamine encapsulated polymeric micelle system(TP/CPA-PM) was prepared and its in-vitro antitumor activities were investigated. Methods: Self-prepared PEG-PLC was used as a carrier material to prepare a novel polymeric micelle system containing both TP and CPA(TP/CPA-PM;TP:CPA = 1:1, w/w) using acetone solvent evaporation method. The micelles were characterized in terms of particle size, analyzed by laser particle size analyzer, entrapment efficiency, determined by high-performance liquid chromatography(HPLC), and in-vitro drug release, determined by combining dialysis method with HPLC. The MTS assay was performed at different time points to determine the inhibitory effect of TP/CPA-PM and free-TP/CPA with different concentrations on the proliferation of three kinds of human pancreatic carcinoma cell lines, namely Miapace-2 cells, Kras cells and HPSC cells. Results: The average particle size of TP/CPA-PM(TP:CPA = 1:1, w/w) was(54.3±2.6) nm(n = 3), and the encapsulation efficiency of TP and CPA was(48.7±7.7%)(n = 3) and(58.9±6.0%)(n = 3), respectively. The drug release curve showed that TP in TP/CPA-PM demonstrated fast-release profile, while CPA in TP/CPAPM indicated sustained-release profile. The release of CPA from micelles was faster at pH 5.2 than at pH 7.4. After 24 h of incubation at 37 °C, 39.0% of CPA was released at pH 7.4, while 58.4% was released at pH 5.2. Compared with free-TP/CPA, TP/CPA-PM significantly inhibited the proliferation of the three kinds of pancreatic cancer cells in a dose-dependent and time-dependent manner. Conclusion: These findings indicate that the combination of triptolid and cyclopamine loaded in polymeric micelles is an effective strategy to treat pancreatic tumors.

关键词： Triptolide cyclopamine polymer micelles encapsulation efficiency in-vitro release profile pancreatic cancer cells

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