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CONSTRUCTION OF RECOMBINANT HSCC-1 ADENOVIRUS VECTOR FOR ORAL CANCER GENE THERAPY (구강암 유전자 치료를 위한 재조합 HSCC-1 아데노바이러스의 개발)

  • Kim, Chang-Hyen;Kim, Jin-Woo;Kim, Myung-Jin;Pyo, Sung-Woon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.2
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    • pp.103-109
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    • 2005
  • In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

Applications of Panax ginseng leaves-mediated gold nanoparticles in cosmetics relation to antioxidant, moisture retention, and whitening effect on B16BL6 cells

  • Jimenez-Perez, Zuly Elizabeth;Singh, Priyanka;Kim, Yeon-Ju;Mathiyalagan, Ramya;Kim, Dong-Hyun;Lee, Myoung Hee;Yang, Deok Chun
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.327-333
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    • 2018
  • Background: Bioactive compounds in plant extracts are able to reduce metal ions to nanoparticles through the process of green synthesis. Panax ginseng is an oriental medicinal herb and an adaptogen which has been historically used to cure various diseases. In addition, the P. ginseng leaves-mediated gold nanoparticles are the value-added novel materials. Its potential as a cosmetic ingredient is still unexplored. The aim of this study was to evaluate the antioxidant, moisture retention and whitening properties of gold nanoparticles (PgAuNPs) in cosmetic applications. Methods: Cell-free experiments were performed to evaluate PgAuNP's antioxidant and moisture retention properties and inhibition activity on mushroom tyrosinase. Furthermore, in vitro cell cytotoxicity was evaluated using normal human dermal fibroblast and murine B16BL6 melanoma cells (B16) after treatment with increasing concentrations of PgAuNPs for 24 h, 48 h, and 72 h. Finally, in vitro cell assays on B16 cells were performed to evaluate the whitening effect of PgAuNPs through reduction of cellular melanin content and tyrosinase activity. Results: In vitro DPPH radical scavenging assay results revealed that PgAuNPs exhibited antioxidant activity in a dose-dependent manner. PgAuNPs exhibited moisture retention capacity and effectively inhibited mushroom tyrosinase. In addition, 3-(4,5-dimethyl-thiazol-2yl)-2,5-diphenyl tetrazolium bromide results revealed that PgAuNPs were not toxic to human dermal fibroblast and B16 cells; in addition, they significantly reduced melanin content, tyrosinase activity, and mRNA expression of melanogenesis-associated transcription factor and tyrosinase in B16 cells. Conclusion: Our study is the first report to provide evidence supporting that P. ginseng leaves-capped gold nanoparticles could be used as multifunctional ingredients in cosmetics.

Antitumor Activity of 7-[2-(N-Isopropylamino)ethyl]-(20s)-camptothecin, CKD602, as a Potent DNA Topoisomerase I Inhibitor

  • Lee, Jun-Hee;Lee, Ju-Mong;Kim, Joon-Kyum;Ahn, Soon-Kil;Lee, Sang-Joon;Kim, Mie-Young;Jew, Sang-Sup;Park, Jae-Gab;Hong, Chung-Il
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.581-590
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    • 1998
  • We developed a novel water-soluble camptothecin analobue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respecitively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to imduce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/ED58) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in <5% tumor bearing mouse. this schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I.

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Preparation of Gold Coated Liposomes for CT Contrast Medium (CT 조영을 위한 금 코팅 리포솜의 제조)

  • Wee, Tae In;Jeon, Ye Won;Cho, Young Jae;Cho, Sung Keun;Ha, Jeung;Lee, Jeong Won;Cho, Sun Hang;Han, Hee Dong;Shin, Byung Cheol
    • Journal of the Korean Chemical Society
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    • v.57 no.5
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    • pp.634-639
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    • 2013
  • The use of medical imaging has been increased for diagnosis of cancer or vessel disease. Among the medical imaging, computed tomography (CT) is one of the popular methods, however, which should need administration of contrast medium. Therefore, we developed gold coated liposomes (GCL) as a contrast medium. To coat gold on the liposomal surface, positive charged liposomes was prepared and then negative Au can coat on the liposomal surface by electronic interaction. The size of GCL was 154.8±9.2 nm and surface charge was 27±3.2 mV, respectively. The morphology of GCL was confirmed by electron microscopy (SEM) and transmission electron microscopy (TEM). The coating efficiency of gold was 18%. Chemical procedure for gold coating on liposomal surface was not toxic for cell cytotoxicity by MTT assay. Finally, we demonstrated attractive CT image for GCL. Taken together, the GCL would be useful for various vessel related disease as a contrast medium.

The interaction of serum albumin with ginsenoside Rh2 resulted in the downregulation of ginsenoside Rh2 cytotoxicity

  • Lin, Yingjia;Li, Yang;Song, Zhi-Guang;Zhu, Hongyan;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.330-338
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    • 2017
  • Background: Ginsenoside Rh2 (G-Rh2) is a ginseng saponin that is widely investigated because of its remarkable antitumor activity. However, the molecular mechanism by which (20S) G-Rh2 triggers its functions and how target animals avoid its cytotoxic action remains largely unknown. Methods: Phage display was used to screen the human targets of (20S) G-Rh2. Fluorescence spectroscopy and UV-visible absorption spectroscopy were used to confirm the interaction of candidate target proteins and (20S) G-Rh2. Molecular docking was utilized to calculate the estimated free energy of binding and to structurally visualize their interactions. MTT assay and immunoblotting were used to assess whether human serum albumin (HSA), bovine serum albumin (BSA), and bovine serum can reduce the cytotoxic activity of (20S) G-Rh2 in HepG2 cells. Results: In phage display, (20S) G-Rh2-beads and (20R) G-Rh2-beads were combined with numerous kinds of phages, and a total of 111 different human complementary DNAs (cDNA) were identified, including HSA which had the highest rate. The binding constant and number of binding site in the interaction between (20S)-Rh2 and HSA were 3.5×105M1 and 1, and those in the interaction between (20S) G-Rh2 and BSA were 1.4×105M1 and 1. The quenching mechanism is static quenching. HSA, BSA and bovine serum significantly reduced the proapoptotic effect of (20S) G-Rh2. Conclusion: HSA and BSA interact with (20S) G-Rh2. Serum inhibited the activity of (20S) G-Rh2 mainly due to the interaction between (20S) G-Rh2 and serum albumin (SA). This study proposes that HSA may enhance (20S) G-Rh2 water solubility, and thus might be used as nanoparticles in the (20S) G-Rh2 delivery process.

Physiological Activities of Phellinus ribis Extracts (찔레 영지버섯(Phellinus ribis) 추출물의 생리활성)

  • Song, Jae-Hwan;Lee, Hyun-Sook;Hwang, Jin-Kook;Chung, Tae-Young;Hong, Sung-Ryul;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.690-695
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    • 2003
  • Physiological activities of 40% ethanol extracts of Phellinus ribis were studied by employing several biological and biochemical assays. The extracts of Phellinus ribis displayed nitrate-scavenging activities (NSA) at pH 1.2 as with 64% NSA with 1.0 mg/mL of the extracts. They also had 91% 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities at the concentration of 1.0 mg/mL. Antioxidant activities of the extracts (at 0.5 mg/mL) on the autoxidation of linoleic acid (p<0.001) were also observed.. The inhibitory effect of the extracts on angiotensin converting enzyme was 11%. Cytotoxic effects of Phellinus ribis extracts against human cancer cell tines were also examined using MTT assay. The extracts (at 50 mg/mL) had severe growth inhibitory effects on A549, Hela, AGS, and SK-Hep-1, which were 8, 44, 76 and 42%, respectively. Ames test indicated that the extracts had no mutating effects on Salmonella typhimurium TA98 and TA100.

Inhibitory Effects of Hoechunyanggyeok-san on Inflammation in Vivo and in Vitro (회춘양격산 물추출물의 항염증효과)

  • Lee, Tae-Ho;Jo, Mi-Jeong;Park, Sook-Jarh;Lee, Jong-Rok;Back, Young-Doo;Kim, Jong-Reol;Kwon, Young-Kyu;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.6
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    • pp.1341-1348
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    • 2009
  • Hoechunyanggyeok-san (HYS) is a traditional oriental herbal medicine widely used for treating inflammatory disorders. Although there are numerous clinical results of HYS reported in the literature of oriental hebal medicine, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of HYS extract (HYSE) in vivo and in vitro. To determine the cytotoxic concentration of HYSE, cell viability was tested by MTT assay. All four doses of HYSE (0.01, 0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. In order to measure NO levels in culture medium, the cells were treated with 1μg/ml of LPS 1h before adding HYSE for 24 h and then culture medium were reacted with Griess reagent. Increased NO production and iNOS expression were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with HYSE. LPS plays a key role in leading to the massive production of pro-inflammatory cytokines such as TNF-α, IL-1β and IL-6 in macrophages. Thus, we next determined the levels of these cytokines. HYSE reduced the elevated production of TNF-α, IL-1β and IL-6 by LPS. Moreover, the effects of HYSE were in a dose-dependent manner. In vivo, histopathological study, HYSE effectively inheefed the increases of hind paw skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. It, therefore, considered that HYSE will be favorably inheefed the acute edematous inanner. In s. These findings showed that HYSE could have anti-inflammatory effects through the reduction of NO and inflammatory cytokines in macrophage. Furthermore, the reduction of carrageenan-induced paw oedema by HYSE helps to understand its actions on inflammatory conditions.

Anti-Obesity Activity of Euptelea Pleiosperma Ethanol Extract (Euptelea pleiosperma 에탄올 추출물의 항비만 활성)

  • Park, Jung Ae;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.336-342
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    • 2015
  • Previously, Euptelea pleiosperma was identified as one of the useful sources containing anti-oxidative and anti-inflammatory activities for the first time in our research group. In this study, anti-obesity effect of E. pleiosperma ethanol extract (EPEE) was evaluated by using a pancreatic lipase enzyme inhibition assay and a cell culture model system. EPEE suppressed effectively pancreatic lipase enzyme activity dose dependently. Furthermore, EPEE significantly suppressed adipocyte differentiation, lipid accumulation, triglyceride contents, and triggered lipolysis activity on 3T3-L1 preadipocytes in a dose-dependent manner without cytotoxicity. Anti-adipogenic effect of EPEE was modulated by cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins α(C/EBPα), C/EBPβ and peroxisome proliferator-activated receptor γ(PPARγ) gene and protein expressions. Taken together, these results provide the important new insight that E. pleiosperma possesses anti-obesity activities such as pancreatic lipase inhibition, anti-adipogenic, and lipolysis effects. It might be utilized as promising sources in the fields of nutraceuticals. The identification of active compounds that confer anti-obesity activity of EPEE might be needed.

Inhibitory effect of Petalonia binghamiae on neuroinflammation in LPS-stimulated microglial cells (LPS에 의해 활성화된 미세아교세포에서 미역쇠 추출물의 신경염증 보호 효과)

  • Park, Jae Hyeon;Kim, Sung Hun;Lee, Sun Ryung
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.25-31
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    • 2017
  • Purpose: Neuroinflammation is mediated by activation of microglia implicated in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease. Inhibition of neuroinflammation may be an effective solution to treat these brain disorders. Petalonia binghamiae is known as a traditional food, based on multiple biological activities such as anti-oxidant and anti-obesity. In present study, the anti-neuroinflammatory potential of Petalonia binghamiae was investigated in LPS-stimulated BV2 microglial cells. Methods: Cell viability was measured by MTT assay. Production of nitric oxide (NO) was examined using Griess reagent. Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was detected by Western blot analysis. Activation of nuclear factor κB (NFκB) signaling was examined by nuclear translocation of NFκB p65 subunit and phosphorylation of IκB. Results: Extract of Petalonia binghamiae significantly inhibited LPS-stimulated NO production and iNOS/COX-2 protein expression in a dose-dependent manner without cytotoxicity. Pretreatment with Petalonia binghamiae suppressed LPS-induced NFκB p65 nuclear translocation and phosphorylation of IκB. Co-treatment with Petalonia binghamiae and pyrrolidine duthiocarbamate (PDTC), an NFκB inhibitor, reduced LPS-stimulated NO release compared to that in PB-treated or PDTC-treated cells. Conclusion: The present results indicate that extract of Petalonia binghamiae exerts anti-neuroinflammation activities, partly through inhibition of NFκB signaling. These findings suggest that Petalonia binghamiae might have therapeutic potential in relation to neuroinflammation and neurodegenerative diseases.

Effect of Gamitonggyu-tang on Secretion of Airway Mucin and Contractility of Tracheal Smooth Muscle (가미통규탕(加味通竅湯)이 호흡기 뮤신 분비 및 기관 평활근 긴장도에 미치는 영향)

  • Lee, Nam-Yeol;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.21 no.3
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    • pp.109-124
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    • 2007
  • Objectives In the present study, the author intended to investigate whether Gamitonggyu-tang (GTT) significantly affects (since the subject is GTT, you need an 's') in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of an airway, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2 for 3 weeks. Effects of orally-administered GTT for 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using enzyme-linked immunosorbent assay (ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GTT to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed.Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase (LDH) release. Also, the effect of GTT on contractility of isolated tracheal smooth muscle was investigated. Results (1) GTT inhibited hypersecretion of in vivo mucin. However, it did not affect the increase the number of goblet cells (2) GTT significantly increased mucin release from cultured HTSE cells, without significant cytotoxicity (3) GTT chiefly affected the 'mucin' secretion and did not affect the secretion of the other releasable glycoproteins with less molecular weight than mucin (4) GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle.Conclusions This result suggests that GTT can increase mucin secretion during short-term treatment (in vitro) whereas it can inihibit hypersecretion of mucin during long-term treatment (in vivo). The author suggests that the effect GTT with their components should be further investigated and it is valuable to find from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway epithelial cells.

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