• Title/Summary/Keyword: HeLa cell lines

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PAMAM Dendrimer Conjugated with N-terminal Oligopeptides of Mouse Fibroblast Growth Factor 3 as a Novel Gene Carrier

  • Jung, Jinwoo;Lee, Jeil;Kim, Tae-Hun;Yang, Bong Suk;Lee, Eunji;Kim, Youn-Joong;Park, Jong-Sang;Choi, Joon Sig
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1036-1042
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    • 2014
  • In this study, we introduced the RRLR oligopeptide sequences on the surface of polyamidoamine (PAMAM) dendrimer and characterized the physical properties and gene carrier activity of the novel polymer using HEK 293, NIH3T3, and HeLa cells. The RRLR peptide sequences were derived from a mouse fibroblast growth factor 3 (FGF3) protein containing a bipartite NLS motif. The entire sequence of FGF3 is RLRRDAGGRGGVYEHLGGAPRRRK and it has two functional sequences RLRR and RRRK at N-terminus and C-terminus, respectively. In particular, PAMAM G4-RRLR conferred enhanced transfection efficiency and lower cytotoxicity compared with those of PEI 25 kDa, PAMAM G4-R, and PAMAM G4 in various cell lines. These results suggest that the introduction of N-terminal oligopeptides of FGF3 on the surface of PAMAM holds promise as an effective non-viral gene delivery carrier for gene therapy.

Antioxidant and cytotoxic activities of curcumin and its analogs: An exploration of structure-activity relationships (Curcumin과 관련 성분들의 산화방지활성과 세포독성 분석 및 구조와 활성 연관성 조사)

  • Lee, Bo-Hyun;Kim, Hee Jeong;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.53 no.4
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    • pp.463-469
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    • 2021
  • Bioactivities of curcumin, a major pigment of Curcuma longa L., have been widely investigated. In this study, the antioxidant and cytotoxic properties of curcumin and its analogs including ferulic acid, dibenzoylmethane (DBM), and tetrahydrocurcumin (THC), and their structure-activity relationships were assessed. Ferulic acid, THC, and curcumin showed strong scavenging activities against several radicals and exhibited considerable lipid peroxidation inhibitory activity. Curcumin showed the strongest cytotoxic activities against HeLa, HCT-116, IEC-6 and INT 407 cells, whereas ferulic acid did not show any cytotoxic effect up to 100 µM against these cell lines. Cytotoxicity of curcumin and THC was significantly enhanced by superoxide dismutase and diminished by N-acetylcysteine. The combination treatment of curcumin and ferulic acid enhanced the cytotoxicity, whereas the combination of curcumin and DBM offset their toxicity. These results suggest that methoxy phenolic and β-diketon moieties are crucial for the antioxidant- and cytototoxic activities of curcumin, respectively.

Multi-Function of a New Bioactive Secondary Metabolite Derived from Endophytic Fungus Colletotrichum acutatum of Angelica sinensis

  • Ramy S. Yehia
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.806-822
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    • 2023
  • In the current study we assessed a new crystallized compound, 5-(1-hydroxybutyl)-4-methoxy-3-methyl-2H-pyran-2-one (C-HMMP), from the endophytic fungus Colletotrichum acutatum residing in the medicinal plant Angelica sinensis for its in vitro antimicrobial, antibiofilm, antioxidant, antimalarial, and anti-proliferative properties. The promising compound was identified as C-HMMP through antimicrobial-guided fraction. The structure of C-HMMP was unambiguously confirmed by 2D NMR and HIRS spectroscopic analysis. Antimicrobial property testing of C-HMMP showed it to be effective against a variety of pathogenic bacteria and fungi with MICs ranging from 3.9 to 31.25 ㎍/ml. The compound displayed excellent antibiofilm activity against C. albicans, S. aureus, and K. pneumonia. Furthermore, the antimalarial and radical scavenging activities of C-HMMP were clearly dosedependent, with IC50 values of 0.15 and 131.2 ㎍/ml. The anti-proliferative activity of C-HMMP against the HepG-2, HeLa, and MCF-7 cell lines in vitro was investigated by MTT assay, revealing notable anti-proliferative activity with IC50 values of 114.1, 90, and 133.6 ㎍/ml, respectively. Moreover, CHMMP successfully targets topoisomerase I and demonstrated beneficial anti-mutagenicity in the Ames test against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF). Finally, the compound inhibited the activity of α-glucosidase and α-amylase with IC50 values of 144.7 and 118.6 ㎍/ml, respectively. To the best of our knowledge, the identified compound C-HMMP was obtained for the first time from C. acutatum of A. sinensis, and this study demonstrated that C-HMMP has relevant biological significance and could provide better therapeutic targets against disease.

Analysis of Nutritional Composition and Antioxidant Activity of Black Chestnut by Aging (숙성에 의한 흑밤의 영양성분 분석과 항산화 활성)

  • Moon Hee Lee;Ji Su Lee;Ji Won Yang;In Beom Kim;Youn-Je Park
    • The Korean Journal of Food And Nutrition
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    • v.37 no.1
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    • pp.30-39
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    • 2024
  • Black chestnut (BC) was obtained through aging of fresh chestnut (FC) at 80℃ for 15 days. Proximate and mineral compositions along with colors of FC and BC were evaluated. With aging, moisture contents decreased by 50%, whereas sugar contents, carbohydrate contents, and calories increased. Contents of minerals (Fe, P, Ca, Na, Mg, K) were significantly higher in FC than in BC, showing an order of Mg < Ca < P < K in both FC and BC. Using a Hunter color system, it was found that lightness (L), redness (a), and yellowness (b) values of FC were higher than those of BC. Antioxidant and cytotoxic activities of hot water and ethanol (50, 80, 100%) extracts prepared from FC and BC were evaluated. Extraction yields were lower with FC than with BC. Among water and ethanol extracts, water extract showed the highest DPPH radical scavenging activity for both FC and BC. IC50 values for ABTS+ radical scavenging activities increased after aging. Cytotoxicities of FC and BC extracts were similar to each other. They were different against various cell lines (3T3, HeLa, and Sarcoma-180). These results suggest that BC could be used as a new processed food using chestnut.

Development of the Functional Beverage Containing the Prunus mume Extracts (매실 추출물을 함유한 기능성 음료 개발)

  • Bae, Ji-Hyun;Kim, Ki-Jin;Kim, Sung-Mi;Lee, Won-Jae;Lee, Sun-Jang
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.713-719
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    • 2000
  • This study was performed to investigate the cytotoxic effect of the Prunus mume extracts containing beverages on the growth of SNU-16 gastric cancer cell, SNU-C2A colon cancer cell, SNU-449 liver cancer cell and HeLa cervical cancer cell. The inhibitory effect on the growth of the cancer cell lines was examined by MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, cytotoxicity test and microscopy. Also this study aimed to compare the changes of blood electrolytes and osmolarity during submaximal exercise for the intake of the Prunus mume extracts containing beverages. 20% Prunus mume extracts containing beverage exhibited the greatest inhibitory effect on the growth of SNU-16 and significantly inhibited at the concentration of $1000\;{\mu}g/mL$ in the MTT assay. Morphological changes in SNU-16 which treated with the same beverage were observed under inverted microscope. The change of blood electrolytes and osmolarity during submaximal exercise showed no significant differences between before and after intake of the beverage in both groups.

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Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus (인유두종바이러스 검출을 위한 상용화된 cDNA 합성 키트의 평가)

  • Yu, Kwangmin;Park, Sunyoung;Chang, Yunhee;Hwang, Dasom;Kim, Geehyuk;Kim, Jungho;Kim, Sunghyun;Kim, Eun-Joong;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.309-315
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    • 2019
  • Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.

Antioxidant and Cytotoxic Activities of Hot Water and Ethanol Extracts From Caesalpinia sappan (소목의 열수 및 에탄올 추출물의 항산화 및 항암활성)

  • Park, Mi-Hye;Kim, Bumsik
    • Food Engineering Progress
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    • v.21 no.3
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    • pp.249-255
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    • 2017
  • Caesalpinia sappan L. is an oriental medicinal plant distributed in the Asia Pacific region including India, Malaysia, and China. The dried heartwood of Caesalpinia sappan has been traditionally used as an analgesic and anti-inflammatory drug. In this study, the effects of extract methods of C. sappan on contents of total polyphenols and flavonoids, antioxidant activity, and cytotoxic activity were evaluated. As a result, hot water extract from C. sappan (CSWE) significantly exhibited contents of total polyphenols and flavonoids (22.6 mg GAE/g and 14.5 mg QE/g) higher than 70% ethanol extract (CSEE) (17.6 mg GAE/g and 13.2 mg QE/g). However, CSEE showed greater antioxidant activity than CSWE in both DPPH and ABTS. Also, the cytotoxicity of C. sappan against three kinds of cancer cell lines was higher in CSEE than in CSWE. These results show that ethanol extract is a better extract method than hot water method to maintain antioxidant and anti-cancer activities.

Effective Antitumor Activity of a Recombinant Vaccinia Virus Expressing Murine Interleukin 4 (인터루킨-4를 발현하는 재조합 백시니아 바이러스에 의한 암성장의 억제)

  • Yoon, Kee-Jung;Jin, Ning-Yi;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.71-78
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    • 1998
  • Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

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UHRF1 Induces Methylation of the TXNIP Promoter and Down-Regulates Gene Expression in Cervical Cancer

  • Kim, Min Jun;Lee, Han Ju;Choi, Mee Young;Kang, Sang Soo;Kim, Yoon Sook;Shin, Jeong Kyu;Choi, Wan Sung
    • Molecules and Cells
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    • v.44 no.3
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    • pp.146-159
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    • 2021
  • DNA methylation, and consequent down-regulation, of tumour suppressor genes occurs in response to epigenetic stimuli during cancer development. Similarly, human oncoviruses, including human papillomavirus (HPV), up-regulate and augment DNA methyltransferase (DNMT) and histone deacetylase (HDAC) activities, thereby decreasing tumour suppressor genes (TSGs) expression. Ubiquitin-like containing PHD and RING finger domain 1 (UHRF1), an epigenetic regulator of DNA methylation, is overexpressed in HPV-induced cervical cancers. Here, we investigated the role of UHRF1 in cervical cancer by knocking down its expression in HeLa cells using lentiviral-encoded short hairpin (sh)RNA and performing cDNA microarrays. We detected significantly elevated expression of thioredoxin-interacting protein (TXNIP), a known TSG, in UHRF1-knockdown cells, and this gene is hypermethylated in cervical cancer tissue and cell lines, as indicated by whole-genome methylation analysis. Up-regulation of UHRF1 and decreased TXNIP were further detected in cervical cancer by western blot and immunohistochemistry and confirmed by Oncomine database analysis. Using chromatin immunoprecipitation, we identified the inverted CCAAT domain-containing UHRF1-binding site in the TXNIP promoter and demonstrated UHRF1 knockdown decreases UHRF1 promoter binding and enhances TXNIP expression through demethylation of this region. TXNIP promoter CpG methylation was further confirmed in cervical cancer tissue by pyrosequencing and methylation-specific polymerase chain reaction. Critically, down-regulation of UHRF1 by siRNA or UHRF1 antagonist (thymoquinone) induces cell cycle arrest and apoptosis, and ubiquitin-specific protease 7 (USP7), which stabilises and promotes UHRF1 function, is increased by HPV viral protein E6/E7 overexpression. These results indicate HPV might induce carcinogenesis through UHRF1-mediated TXNIP promoter methylation, thus suggesting a possible link between CpG methylation and cervical cancer.

Physiological Activity of Sarcodon aspratus Extracts (능이버섯(Sarcodon aspratus) 추출물의 생리활성)

  • 송재환;이현숙;황진국;한정환;노정근;금동혁;박기문
    • Food Science of Animal Resources
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    • v.23 no.2
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    • pp.172-179
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    • 2003
  • This study was carried out to find the preventive medical and therapeutic effects of Sarcodon asparatus on adult disease by employing several biological and biochemical assays. Nitrate scavenging ability(NSA) of Sarcodan asparatus extracts was displayed up to 99.9% at pH 1.2 in a dose-dependent manner. They also had 90.4% electron donating ability(EDA) at the concentration of 0.1 mg/mL. Extracts of Sarcodon asparatus were also able to function as a powerful antioxidant at all concentrations(0.01∼l.0 mg/mL). Furthermore, we observed that 1 mg/mL concentration of the extracts was more powerful than BHT, With respect to fibrolytic activity, Sarcodon asparatus showed 1,843.8 unit/g, which was higher than streptokinase(1,189 unit/g). The inhibitory effects of the extracts on angiotensin converting enzyme, measured by the normal and pretreatment methods, were 53 and 58%, respectively. We also performed cytotoxicity effect of Sarcodon asparatus extracts on a various cancer cell lines. The growth inhibitory effects of the extracts(5.0 mg/mL) on A549, HeLa, AGS, and SK-Hep-1 cells were 78.9, 55.3, 69.0, and 42.5 %, respectively. Interestingly, Sarcodon asparatusextracts induced mutation on Salmonella typhimurium TA98 and TA100 when Ames test was done.