• Title/Summary/Keyword: in vitro cancer research

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The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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Effects of FasL Expression in Oral Squamous Cell Cancer

  • Fang, Li;Sun, Lin;Hu, Fang-Fang;Chen, Qiao-Er
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.281-285
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    • 2013
  • Purpose: To probe the role of FasL in cell apoptosis in oral squamous cell carcinomas (OSCCs). Methods: The expression of Fas/FasL was assessed in 10 cases of normal oral epithelium, 38 cases of OSCC and tumor infiltrating lymphocytes (TIL), and 11 cases of metastatic lymph nodes by immunohistochemistry. Apoptosis of tumor cells and TIL was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). FasL-induction of T cell apoptosis was tested by co-culture assay in vitro with SCC-9 and Jurkat T cells. Results: The 10 cases of normal oral epithelium all demonstrated extensive expression of Fas, the positive rate being largely down-regulated in OSCC (21/38) (P<0.05) compared to the normal (10/10). At the same time, the positive rate of FasL significantly increased in OSCC (P<0.05) especially those with lymph node metastasis (P<0.05). The positive rates of Fas in well and middle differentiated OSCC were higher than those in poor differentiated OSCC (P<0.05). The AI of tumor cells in Fas-positive OSCC was remarkably higher than that in Fas-negative OSCC (P<0.01), with a positive correlation between Fas expression and cell differentiation as well as apoptosis (r=0.68, P<0.01). The AI of tumor cells in FasL positive OSCC was remarkably lower than that in control while the AI of TIL was higher than in FasL negative OSCC (P<0.05). The AI of tumor cells reversely correlated with that of TIL (r = -0. 72, P<0.05). It was found that SCC-9 cells expressing functional FasL could induce apoptosis of Jurkat cells as demonstrated by co-culture assays. As a conclusion, it is evident that OSCC cells expressing FasL can induce apoptosis in Fas-expressing T cells. Conclusions: In progression of OSCC, expression of the Fas/FasL changes significantly. The results suggest that FasL is a mediator of immune privilege in OSCC and may serve as an marker for predicting malignant change in oral tissues.

The Production of Anti-cancer Substances by in vitro Grown Cultures of Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Park, Kyung-Hwa;Kim, Yong-Hae;Yoon, Eui-Soo;Kang, Tae-Jin;Park, Kwang-Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 1999.10a
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    • pp.46-57
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    • 1999
  • Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

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Santalum album Linn wood and its oil: An aromatic Unani traditional medicine with versatile pharmacological activities

  • Sultana, Arshiya;Rahman, Khaleequr
    • CELLMED
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    • v.8 no.3
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    • pp.14.1-14.6
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    • 2018
  • Santalum album Linn. [Family: Santalaceae] is commonly known as white sandalwood, sandal safaid and safed chandan. It is one of the most valuable trees and second costliest wood in the world. Sandalwood and its oil is extensively used in the Unani and other traditional systems of medicine as it has blood purifier, anti-inflammatory, analgesic, exhilarant, cardiotonic, antiseptic, nervine tonic and expectorant properties. It is used in skin, cardiac, liver, gastrointestinal, respiratory, integument and urogenital disorders. These uses are supported and proven by many in vitro or in vivo studies. The proven pharmacological activities of S. album are antimicrobial, anti-oxidant, anti-inflammatory, antimutagenic and anti-fatigue. The research has proven that sandal oil or its constituents have anti-microbial activity. Sandalwood oil showed skin cancer preventive effect in mice and its constituent alpha santalol showed the anticancer property. The methanolic extract of wood was confirmed for antioxidant, free radical scavenging, analgesic and anti-inflammatory activities. ${\alpha}$ and ${\beta}$ santalols present in sandal oil showed sedative effects. Sandalwood tea had a significant effect on heart muscles of frog and showed increased myocardial contractility. Its oil showed significant changes in hepatic xenobiotic metabolizing enzymes. Sandalwood oil and its major constituents showed less acute oral and dermal toxicity in laboratory animals. Hence, the aforementioned studies justify the uses of sandalwood and its oil mentioned in the classical Unani literature. However, further clinical trials are suggested to confirm its efficacy and safety in humans.

Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes LipofectamineTM2000

  • Huang, Fei;Zhao, Feng;Liang, Li-Ping;Zhou, Mei;Qu, Zhi-Ling;Cao, Yan-Zhen;Lin, Chen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7749-7754
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    • 2015
  • Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: $Lipofectamine^{TM}2000$ as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are $1.5{\times}10^4$ (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P<0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

Medicinal aspects of Murraya koenigii mediated silver nanoparticles

  • Mumtaz, Sumaira;Nadeem, Raziya;Sarfraz, Raja A.;Shahid, Muhammad
    • Advances in nano research
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    • v.11 no.6
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    • pp.657-665
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    • 2021
  • The present work aimed to explore green approach via aqueous leaves extract of Murraya koenigii (ALEMk) for the synthesis of silver nanoparticles (AgNPsMk) in single step. The synthesis process was visualized with a color change and monitored by employing UV/Visible spectroscopy and a clear peak attained at 420 nm confirming the synthesis of AgNPsMk. The possible functional groups present in the extract which participated in the synthesis of AgNPsMk were identified with the help of FTIR spectroscopy. Further characterization using TEM images revealed the spherical shape of AgNPsMk with average particle size of 20 nm displaying well dispersion throughout the solution. Pronounced antioxidant activities of AgNPsMk at increased concentrations observed which evidencing strong radical scavenging ability. Moreover, AgNPsMk exhibited strong antibacterial behavior when tested against bacterial strains of Escherichia coli and Bacillus subtilis. Moving ahead, in vitro cytotoxicity work revealed potent cell viability loss appearing in AU565 and HeLa cancer cell lines on exposure to AgNPsMk at increased concentration. Finally, in vivo assessment carried out inside male Wistar rats indicated non toxic effect on examined liver tissues besides biochemical analysis including bilirubin, alkaline phosphtase (ALP) and serum glutamate pyruvate transaminase (SGPT) which found within the normal range when compared with control. The prior research work profoundly apprises the potential of green synthesized AgNPsMk to play a significant role in biomedical applications and formulations.

Induction of cardiomyocyte-like cells from hair follicle cells in mice

  • Yong-Hee Kim;Bang-Jin Kim;Seok-Man Kim;Sun-Uk Kim;Buom-Yong Ryu
    • International Journal of Molecular Medicine
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    • v.43 no.5
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    • pp.2230-2240
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    • 2019
  • Hair follicles (HFs) are a well-characterized niche for adult stem cells (SCs), and include epithelial and melanocytic SCs. HF cells are an accessible source of multipotent adult SCs for the generation of the interfollicular epidermis, HF structures and sebaceous glands in addition to the reconstitution of novel HFs in vivo. In the present study, it was demonstrated that HF cells are able to be induced to differentiate into cardiomyocyte-like cells in vitro under specific conditions. It was determined that HF cells cultured on OP9 feeder cells in KnockOut-Dulbecco's modified Eagle's medium/B27 in the presence of vascular endothelial growth factors differentiated into cardiomyocyte-like cells that express markers specific to cardiac lineage, but do not express non-cardiac lineage markers including neural stem/progenitor cell, HF bulge cells or undifferentiated spermatogonia markers. These cardiomyocyte-like cells exhibited a spindle- and filament-shaped morphology similar to that presented by cardiac muscles and exhibited spontaneous beating that persisted for over 3 months. These results demonstrate that SC reprogramming and differentiation may be induced without resulting in any genetic modification, which is important for the clinical applications of SCs including tissue and organ regeneration.

Anti-Angiogenic Activity of Acalycixenolide E, a Novel Marine Natural Product from Acallycigorgia inermis

  • Kwon, Ho-Jeong;Kim, Jin-Hee;Jung, Hye-Jin;Kwon, Yong-Guen;Kim, Min-Young;Rho, Jung-Rae;Shin, Jong-Heon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.656-662
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    • 2001
  • Angiogenesis is known as a crucial process in the growth and spreading of tumor cells. Accordingly, the effective inhibition of this process would appear to be a promising way to cure angiogenesis-related diseases, including cancer. This study demonstrates that acalycixenolide E (AX-E) from the marine organism Acalycigorgia inermis exhibits a potent anti-angiogenic activity both in vitro and in vivo. AX-E inhibits the bFGF-induced proliferation of HUVECs in a dose dependent manner, along with the bFGF-induced migration, invasion, and tube formation of HUVECs. Moreover, AX-E potently inhibits the in vivo neovascularization of the chorioallantoic membranes (CAMs) of growing chick embryos. interestingly, AX-E suppresses the expression of metalloproteases 2 and 9, yet shows no effect on their activities. The novel chemical structure and potent anti-angiogenic activity of AX-E will be of great value in elucidating the molecular mechanism of angiogenesis as well as in the development of a novel anti-angiogenic drug.

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Effects of Mollugin on Hepatic Cytochrome P450 in Male ICR Mice as Determined by Liquid Chromatography/Tandem Mass Spectrometry

  • Song, Min;Hong, Miri;Choi, Hyun Gyu;Jahng, Yurngdong;Lee, Seung Ho;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • v.3 no.4
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    • pp.104-107
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    • 2012
  • Mollugin isolated from Rubia cordifolia is known to have anti-inflammatory, anti-cancer, and anti-viral activities. In the present study, a cocktail probe assay and LC-MS/MS were used to investigate the modulating effect of mollugin on cytochrome P450 (CYP) enzymes in male ICR mice. After mollugin was orally administrated to mice at the 20, 40, or 80 mg/kg for 3 days, the activities of CYP in hepatic S-9 fractions were investigated. Unlike the selective inhibitory effect of mollugin on CYP1A2-catalyzed phenacetin O-deethylation in vitro, mollugin only significantly inhibited the activity of CYP2E1-catalyzed chlorzoxazone 6-hydroxylase in vivo. The activities of other CYPs were only slightly altered by mollugin. The results of this study suggest that mollugin might cause herb-drug interactions via the selective inhibition of CYP2E1 in vivo.

Synthesis, Characterization and in vitro Anti-Tumoral Evaluation of Erlotinib-PCEC Nanoparticles

  • Barghi, Leila;Asgari, Davoud;Barar, Jaleh;Nakhlband, Aylar;Valizadeh, Hadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10281-10287
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    • 2015
  • Background: Development of a nanosized polymeric delivery system for erlotinib was the main objective of this research. Materials and Methods: Poly caprolactone-polyethylene glycol-polycaprolactone (PCEC) copolymers with different compositions were synthesized via ring opening polymerization. Formation of triblock copolymers was confirmed by HNMR as well as FT-IR. Erlotinib loaded nanoparticles were prepared by means of synthesized copolymers with solvent displacement method. Results: Physicochemical properties of obtained polymeric nanoparticles were dependent on composition of used copolymers. Size of particles was decreased with decreasing the PCL/PEG molar ratio in used copolymers. Encapsulation efficiency of prepared formulations was declined by decreasing their particle size. Drug release behavior from the prepared nanoparticles exhibited a sustained pattern without a burst release. From the release profiles, it can be found that erlotinib release rate from polymeric nanoparticles is decreased by increase of CL/PEG molar ratio of prepared block copolymers. Based on MTT assay results, cell growth inhibition of erlotinib has a dose and time dependent pattern. After 72 hours of exposure, the 50% inhibitory concentration (IC50) of erlotinib hydrochloride was appeared to be $14.8{\mu}M$. Conclusions: From the obtained results, it can be concluded that the prepared PCEC nanoparticles in this study might have the potential to be considered as delivery system for erlotinib.