• Title/Summary/Keyword: Key chain

Search Result 799, Processing Time 0.024 seconds

Policies to Manage Drug Shortages in Selected Countries: A Review and Implications (주요국의 수급불안정 의약품 관리제도에 관한 고찰과 한국에의 시사점)

  • Inmyung Song;Sang Jun Jung;Eunja Park;Sang-Eun Choi;Eun-A Lim;Sanghyun Kim;Dongsook Kim
    • Health Policy and Management
    • /
    • v.34 no.2
    • /
    • pp.106-119
    • /
    • 2024
  • Drug shortage is a persistent phenomenon that poses a public health risk worldwide and occurs due to a range of causes. The purpose of this study is to review key policies to prepare for and respond to drug shortages in selected countries, such as the United States, Canada, and some European countries in order to draw implications. This study reviewed the reports and articles derived from search engines and Google Scholar by using keywords such as drug shortage and stock-out. Over the last decade or so, the United States have strengthened requirements on advance notification for disruption and interruption of drug manufacturing, established the Inter-agency Drug Shortages Task Force to promote the communication and coordination of responses, and expedited drug regulatory processes. Similarly, Canada established the Multi-Stakeholder Steering Committee on drug shortages by involving representatives from central and local governments and private sectors. Canada also adopted a tiered approach to the communication of drug shortages based on the assessment of the severity of the shortage problem and released a detailed information guide on communication. In 2019, the joint task force between the European Medicines Agency and the Heads of Medicines Agencies issued guidelines on drug shortage communication in the European Economic Area. The countries reviewed in this paper focus on communication across different stakeholders for the monitoring of and timely response to drug shortages. The efforts to protect public health from the negative impact of the drug shortage crisis would require multi-sectorial and multi-governmental coordination and development of guidelines.

Design of Authentication Mechinism for Command Message based on Double Hash Chains (이중 해시체인 기반의 명령어 메시지 인증 메커니즘 설계)

  • Park Wang Seok;Park Chang Seop
    • Convergence Security Journal
    • /
    • v.24 no.1
    • /
    • pp.51-57
    • /
    • 2024
  • Although industrial control systems (ICSs) recently keep evolving with the introduction of Industrial IoT converging information technology (IT) and operational technology (OT), it also leads to a variety of threats and vulnerabilities, which was not experienced in the past ICS with no connection to the external network. Since various control command messages are sent to field devices of the ICS for the purpose of monitoring and controlling the operational processes, it is required to guarantee the message integrity as well as control center authentication. In case of the conventional message integrity codes and signature schemes based on symmetric keys and public keys, respectively, they are not suitable considering the asymmetry between the control center and field devices. Especially, compromised node attacks can be mounted against the symmetric-key-based schemes. In this paper, we propose message authentication scheme based on double hash chains constructed from cryptographic hash function without introducing other primitives, and then propose extension scheme using Merkle tree for multiple uses of the double hash chains. It is shown that the proposed scheme is much more efficient in computational complexity than other conventional schemes.

Effect of Various Pathological Conditions on Nitric Oxide Level and L-Citrulline Uptake in Motor Neuron-Like (NSC-34) Cell Lines

  • Shashi Gautam;Sana Latif;Young-Sook Kang
    • Biomolecules & Therapeutics
    • /
    • v.32 no.1
    • /
    • pp.154-161
    • /
    • 2024
  • Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive paralysis. L-Citrulline is a nonessential neutral amino acid produced by L-arginine via nitric oxide synthase (NOS). According to previous studies, the pathogenesis of ALS entails glutamate toxicity, oxidative stress, protein misfolding, and neurofilament disruption. In addition, L-citrulline prevents neuronal cell death in brain ischemia; therefore, we investigated the change in the transport of L-citrulline under various pathological conditions in a cell line model of ALS. We examined the uptake of [14C]L-citrulline in wild-type (hSOD1wt/WT) and mutant NSC-34/ SOD1G93A (MT) cell lines. The cell viability was determined via MTT assay. A transport study was performed to determine the uptake of [14C]L-citrulline. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to determine the expression levels of rat large neutral amino acid transported 1 (rLAT1) in ALS cell lines. Nitric oxide (NO) assay was performed using Griess reagent. L-Citrulline had a restorative effect on glutamate induced cell death, and increased [14C]L-citrulline uptake and mRNA levels of the large neutral amino acid transporter (LAT1) in the glutamate-treated ALS disease model (MT). NO levels increased significantly when MT cells were pretreated with glutamate for 24 h and restored by co-treatment with L-citrulline. Co-treatment of MT cells with L-arginine, an NO donor, increased NO levels. NSC-34 cells exposed to high glucose conditions showed a significant increase in [14C]L-citrulline uptake and LAT1 mRNA expression levels, which were restored to normal levels upon co-treatment with unlabeled L-citrulline. In contrast, exposure of the MT cell line to tumor necrosis factor alpha, lipopolysaccharides, and hypertonic condition decreased the uptake significantly which was restored to the normal level by co-treating with unlabeled L-citrulline. L-Citrulline can restore NO levels and cellular uptake in ALS-affected cells with glutamate cytotoxicity, pro-inflammatory cytokines, or other pathological states, suggesting that L-citrulline supplementation in ALS may play a key role in providing neuroprotection.

A Study on Impact of Factors Influencing Maritime Freight Rates Using Poisson and Negative Binomial Regression Analysis on Blank Sailings of Shipping Companies (포아송 및 음이항 회귀분석을 이용한 해상운임 결정요인이 해운선사의 블랭크 세일링에 미치는 영향 분석 연구)

  • Won-Hyeong Ryu;Hyung-Sik Nam
    • Journal of Navigation and Port Research
    • /
    • v.48 no.1
    • /
    • pp.62-77
    • /
    • 2024
  • In the maritime shipping industry, imbalance between supply and demand has persistently increased, leading to the utilization of blank sailings by major shipping companies worldwide as a key means of flexibly adjusting vessel capacity in response to shipping market conditions. Traditionally, blank sailings have been frequently implemented around the Chinese New Year period. However, due to unique circumstances such as the global pandemic starting in 2020 and trade tensions between the United States and China, shipping companies have recently conducted larger-scale blank sailings compared to the past. As blank sailings directly impact freight transport delays, they can have negative repercussions from perspectives of both businesses and consumers. Therefore, this study employed Poisson regression models and negative binomial regression models to analyze the influence of maritime freight rate determinants on shipping companies' decisions regarding blank sailings, aiming to proactively address potential consequences. Results of the analysis indicated that, in Poisson regression analysis for 2M, significant variables included global container shipping volume, container vessel capacity, container ship scrapping volume, container ship newbuilding index, and OECD inflation. In negative binomial regression analysis, ocean alliance showed significance with global container shipping volume and container ship order volume, the alliance with container ship capacity and interest rates, non-alliance with international oil prices, global supply chain pressure index, container ship capacity, OECD inflation, and total alliance with container ship capacity and interest rates.

A Study on the Competitive Factor of Global Logistics Hub Cities Using a Importance-Performance Analysis : Focusing on the Case of Incheon Metropolitan City (IPA분석을 통한 글로벌 물류 허브도시 경쟁요인에 관한 연구 : 인천광역시 사례를 중심으로)

  • Lee, Myeong-Hwa;Shin, Mi-Na;Kim, Un-Soo
    • Journal of Korea Port Economic Association
    • /
    • v.40 no.2
    • /
    • pp.205-219
    • /
    • 2024
  • This study assesses Incheon Metropolitan City's potential as a global logistics hub amid intensified competition since the 2000s. Utilizing Importance-Performance Analysis(IPA), it evaluates competitive factors for logistics hub cities and Incheon's current positioning. The research identifies world-class infrastructure development and global city connectivity as key competitiveness factors. While Incheon, with its international airport and port, currently functions as a logistics hub, areas for improvement emerge. Recommendations include developing specialized cargo infrastructure for cold-chain and e-commerce, expanding the global network through multimodal transportation, and addressing gaps in smart and eco-friendly logistics. These suggestions encompass professional training, information platform establishment, and sector-wide decarbonization initiatives. The study's significance lies in its IPA-driven evaluation of competitiveness factors and Incheon's status, providing actionable recommendations for strategic planning to enhance the city's position as a global logistics hub.

Structural analysis, anti-inflammatory activity of the main water-soluble acidic polysaccharides (AGBP-A3) from Panax quinquefolius L berry

  • Zhihao Zhang;Huijiao Yan;Hidayat Hussain;Xiangfeng Chen;Jeong Hill Park;Sung Won Kwon;Lei Xie;Bowen Zheng;Xiaohui Xu;Daijie Wang;Jinao Duan
    • Journal of Ginseng Research
    • /
    • v.48 no.5
    • /
    • pp.454-463
    • /
    • 2024
  • Background: Panax quinquefolius L, widely recognized for its valuable contributions to medicine, has aroused considerable attention globally. Different from the extensive research has been dedicated to the root of P. quinquefolius, its berry has received relatively scant focus. Given its promising medicinal properties, this study was focused on the structural characterizations and anti-inflammatory potential of acidic polysaccharides from the P. quinquefolius berry. Materials and methods: P. quinquefolius berry was extracted with hot water, precipitated by alcohol, separated by DEAE-52-cellulose column to give a series of fractions. One of these fractions was further purified via Sephadex G-200 column to give three fractions. Then, the main fraction named as AGBP-A3 was characterized by methylation analysis, NMR spectroscopy, etc. Its anti-inflammatory activity was assessed by RAW 264.7 cell model, zebrafish model and molecular docking. Results: The main chain comprised of α-L-Rhap, α-D-GalAp and β-D-Galp, while the branch consisted mainly of α-L-Araf, β-D-Glcp, α-D-GalAp, β-D-Galp. The RAW264.7 cell assay results showed that the inhibition rates against IL-6 and IL-1β secretion at the concentration of 625 ng/mL were 24.83 %, 11.84 %, while the inhibition rate against IL-10 secretion was 70.17 % at the concentration of 312 ng/mL. In the zebrafish assay, the migrating neutrophils were significantly reduced in number, and their migration to inflammatory tissues was inhibited. Molecular docking predictions correlated well with the results of the anti-inflammatory assay. Conclusion: The present study demonstrated the structure of acidic polysaccharides of P. quinquefolius berry and their effect on inflammation, providing a reference for screening anti-inflammatory drugs.

Relationship among porcine lncRNA TCONS_00010987, miR-323, and leptin receptor based on dual luciferase reporter gene assays and expression patterns

  • Ding, Yueyun;Qian, Li;Wang, Li;Wu, Chaodong;Li, DengTao;Zhang, Xiaodong;Yin, Zongjun;Wang, Yuanlang;Zhang, Wei;Wu, Xudong;Ding, Jian;Yang, Min;Zhang, Liang;Shang, Jinnan;Wang, Chonglong;Gao, Yafei
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.33 no.2
    • /
    • pp.219-229
    • /
    • 2020
  • Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORTTM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

Retail Product Development and Brand Management Collaboration between Industry and University Student Teams (산업여대학학생단대지간적령수산품개발화품패관리협작(产业与大学学生团队之间的零售产品开发和品牌管理协作))

  • Carroll, Katherine Emma
    • Journal of Global Scholars of Marketing Science
    • /
    • v.20 no.3
    • /
    • pp.239-248
    • /
    • 2010
  • This paper describes a collaborative project between academia and industry which focused on improving the marketing and product development strategies for two private label apparel brands of a large regional department store chain in the southeastern United States. The goal of the project was to revitalize product lines of the two brands by incorporating student ideas for new solutions, thereby giving the students practical experience with a real-life industry situation. There were a number of key players involved in the project. A privately-owned department store chain based in the southeastern United States which was seeking an academic partner had recognized a need to update two existing private label brands. They targeted middle-aged consumers looking for casual, moderately priced merchandise. The company was seeking to change direction with both packaging and presentation, and possibly product design. The branding and product development divisions of the company contacted professors in an academic department of a large southeastern state university. Two of the professors agreed that the task would be a good fit for their classes - one was a junior-level Intermediate Brand Management class; the other was a senior-level Fashion Product Development class. The professors felt that by working collaboratively on the project, students would be exposed to a real world scenario, within the security of an academic learning environment. Collaboration within an interdisciplinary team has the advantage of providing experiences and resources beyond the capabilities of a single student and adds "brainpower" to problem-solving processes (Lowman 2000). This goal of improving the capabilities of students directed the instructors in each class to form interdisciplinary teams between the Branding and Product Development classes. In addition, many universities are employing industry partnerships in research and teaching, where collaboration within temporal (semester) and physical (classroom/lab) constraints help to increase students' knowledge and experience of a real-world situation. At the University of Tennessee, the Center of Industrial Services and UT-Knoxville's College of Engineering worked with a company to develop design improvements in its U.S. operations. In this study, Because should be lower case b with a private label retail brand, Wickett, Gaskill and Damhorst's (1999) revised Retail Apparel Product Development Model was used by the product development and brand management teams. This framework was chosen because it addresses apparel product development from the concept to the retail stage. Two classes were involved in this project: a junior level Brand Management class and a senior level Fashion Product Development class. Seven teams were formed which included four students from Brand Management and two students from Product Development. The classes were taught the same semester, but not at the same time. At the beginning of the semester, each class was introduced to the industry partner and given the problem. Half the teams were assigned to the men's brand and half to the women's brand. The teams were responsible for devising approaches to the problem, formulating a timeline for their work, staying in touch with industry representatives and making sure that each member of the team contributed in a positive way. The objective for the teams was to plan, develop, and present a product line using merchandising processes (following the Wickett, Gaskill and Damhorst model) and develop new branding strategies for the proposed lines. The teams performed trend, color, fabrication and target market research; developed sketches for a line; edited the sketches and presented their line plans; wrote specifications; fitted prototypes on fit models, and developed final production samples for presentation to industry. The branding students developed a SWOT analysis, a Brand Measurement report, a mind-map for the brands and a fully integrated Marketing Report which was presented alongside the ideas for the new lines. In future if the opportunity arises to work in this collaborative way with an existing company who wishes to look both at branding and product development strategies, classes will be scheduled at the same time so that students have more time to meet and discuss timelines and assigned tasks. As it was, student groups had to meet outside of each class time and this proved to be a challenging though not uncommon part of teamwork (Pfaff and Huddleston, 2003). Although the logistics of this exercise were time-consuming to set up and administer, professors felt that the benefits to students were multiple. The most important benefit, according to student feedback from both classes, was the opportunity to work with industry professionals, follow their process, and see the results of their work evaluated by the people who made the decisions at the company level. Faculty members were grateful to have a "real-world" case to work with in the classroom to provide focus. Creative ideas and strategies were traded as plans were made, extending and strengthening the departmental links be tween the branding and product development areas. By working not only with students coming from a different knowledge base, but also having to keep in contact with the industry partner and follow the framework and timeline of industry practice, student teams were challenged to produce excellent and innovative work under new circumstances. Working on the product development and branding for "real-life" brands that are struggling gave students an opportunity to see how closely their coursework ties in with the real-world and how creativity, collaboration and flexibility are necessary components of both the design and business aspects of company operations. Industry personnel were impressed by (a) the level and depth of knowledge and execution in the student projects, and (b) the creativity of new ideas for the brands.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
    • /
    • 2016.05a
    • /
    • pp.19-20
    • /
    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

  • PDF

The Change of Cell-cycle Related Proteins and Tumor Suppressive Effect in Non-small Cell Lung Cancer Cell Line after Transfection of p16(MTS1) Gene (폐암세포에 p16 (MTS1) 유전자 주입후 암생성능의 변화 및 세포주기관련 단백질의 변동에 관한 연구)

  • Kim, Young-Whan;Kim, Jae-Yeol;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
    • /
    • v.44 no.4
    • /
    • pp.796-805
    • /
    • 1997
  • Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

  • PDF