• BMB Reports
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• 제56권11호
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• pp.612-617
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• 2023

Pleiotropic regulator 1 (PLRG1), a highly conserved element in the spliceosome, can form a NineTeen Complex (NTC) with Prp19, SPF27, and CDC5L. This complex plays crucial roles in both pre-mRNA splicing and DNA repair processes. Here, we provide evidence that PLRG1 has a multifaceted impact on cancer cell proliferation. Comparing its expression levels in cancer and normal cells, we observed that PLRG1 was upregulated in various tumor tissues and cell lines. Knockdown of PLRG1 resulted in tumor-specific cell death. Depletion of PLRG1 had notable effects, including mitotic arrest, microtubule instability, endoplasmic reticulum (ER) stress, and accumulation of autophagy, ultimately culminating in apoptosis. Our results also demonstrated that PLRG1 downregulation contributed to DNA damage in cancer cells, which we confirmed through experimental validation as DNA repair impairment. Interestingly, when PLRG1 was decreased in normal cells, it induced G1 arrest as a self-protective mechanism, distinguishing it from effects observed in cancer cells. These results highlight multifaceted impacts of PLRG1 in cancer and underscore its potential as a novel anti-cancer strategy by selectively targeting cancer cells.

• 생명과학회지
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• 제28권4호
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• pp.389-397
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• 2018

당 단백질에 붙어 있는 당사슬 구조는 형질전환 돼지 유즙으로 분비되는 의약용 단백질의 생물학적 활성, 안정성 그리고 안전성에 영향을 줄 수 있다. 형질전환 동물을 이용한 치료용 당 단백질 생산은 유선 세포에서 이루어지는 당사슬 부가능력에 의해 제한되며, 균일한 당사슬 형태를 가지는 당 단백질 생산은 도전 과제로 남아있다. ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) 유전자는 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 단백질 당화기작에 중요한 효소이지만, 돼지 당 전이효소에 대한 정보는 매우 제한적이다. 따라서, 돼지 B3GNT1 (pB3GNT1) 유전자를 클로닝하고 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 기능적 특성을 조사하였다. 몇가지 다른 프라이머를 사용하여 전체 전사영역(ORF)을 함유하는 부분적인 pB3GNT1 mRNA 염기서열을 간 조직으로부터 분리하였다. 클로닝 된 pB3GNT1의 ORF는 1,248개의 뉴클레오티드를 가지며, 415개 아미노산 잔기로 구성되어 있었다. pB3GNT1 유전자의 장기별 발현특성은 성돈 및 자돈의 여러 기관에서 분석하였다. pB3GNT1 mRNA 발현 수준은 심장, 소장 보다는 근육에서 높았지만 폐에서는 낮았다. pB3GNT1의 기능적 특성 분석을 위해 돼지 신장 세포주(PK-15)에서 pB3GNT1 유전자의 안정적인 발현을 확립하였다. 그 결과, PK-15 세포에서 pB3GNT1 발현에 의한 당화 패턴은 총 시알산 증가에는 영향을 미치지 않지만, poly-N-아세틸글루코사민은 증가하는 것으로 나타났다. 본 연구는 생물반응기로 형질전환 돼지를 이용할 때 희망하는 당사슬을 부가하여 치료 가능성을 높이며 개선된 활성을 나타내는 당단백질 생산에 도움이 될 것이다.

• 한국약용작물학회지
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• 제23권3호
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• pp.190-197
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• 2015

In this study, we tried to offer the possibility of White Hibiscus syriacus L. (WHS) flower extracts as a preventive and improving agent of osteoporosis that bone mass reduction is induced by an decrease of osteoblast involved in bone formation and increase of bone resorption by osteoclast activity. As a results, it was found to have antioxidant activity and contain a flavonoid contents ($47.74 mg/g) of the WHS flower. There was cytotoxicity at more than $250{\mu}g/m{\ell}$ concentration of WHS flower extract of RANKL-induced osteoclast in RAW264.7. There were no significant inhibited TRAP activity in the WHS leaf and stem. However, it was confirmed that it is significantly inhibited the differentiation activity of osteoclasts in 50 and $100{\mu}g/m{\ell}$ concentration of cells of stability levels of only WHS flower extracts (p < 0.01). The WHS flower prominently inhibited RANKL-induced osteoclast differentiation activity by decreased calcitonin receptor and TRAP mRNA (p < 0.01). These results indicate that of osteoclasts differentiation activity is inhibited by protection of oxidative stress due to the antioxidant activity of the WHS flower. Therefore, suggesting the WHS flower may be a presents the possibility as a preventive and therapeutic agents for osteoporosis.

• 대한화장품학회지
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• 제44권2호
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• pp.171-181
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• 2018

제니스테인(genistein)은 대두에서 추출한 대표적인 이소 플라본 화합물 중 하나이며 노화 방지 및 항염증 활성 효과에 대한 연구가 많이 이뤄졌다. 그러나 제니스테인은 유기용매에 높은 용해도를 보일지라도 물에 대한 수용성은 매우 낮아 생체이용률이 떨어진다. 따라서 본 연구에서는 제니스테인의 수용성과 안정성이 크게 향상된 제니스테인 cyclodextrin 포접체(genistein CD complex)를 제니스테인과 직접 비교 분석하고자 하였다. 우선 세포독성 실험을 위해 RAW264.7 대식세포를 대상으로 CCK-8 assay를 시행하였고, 제니스테인 및 제니스테인 cyclodextrin 포접체 모두 $10{\mu}g/mL$ 농도부터 세포독성이 나타나 최대 농도는 $10{\mu}g/mL$로 설정하고 실험을 진행하였다. LPS에 의해 활성화 된 RAW264.7 세포에서 NO(nitric oxide) 생성 및 iNOS mRNA 발현을 관찰한 결과 제니스테인 CD 포접체가 제니스테인 자체 보다 더 효과적으로 억제하였다. 또한 $IL1-{\alpha}$, $IL1-{\beta}$, IL-6 및 $TNF-{\alpha}$와 같은 염증성 사이토카인의 mRNA 발현이 농도 의존적으로 감소됨을 확인하였다. 이 뿐 아니라 인간 각질형성세포인 HaCaT 세포를 이용해 TEER 및 피부장벽 강화 효과를 관찰한 결과 제니스테인 CD 포접체 처리군에서 TEER이 농도 의존적으로 증가되었고, 세포 이동 실험에서도 동일한 결과를 얻을 수 있었다. 따라서 제니스테인 CD 포접체에 대한 피부 재생 및 장벽 강화에 관한 임상 연구등이 수행된다면, 효과적인 아토피 피부염 또는 피부장벽 개선 기능성 화장품 원료로 사용될 수 있을 것으로 기대된다.

• Molecules and Cells
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• 제22권3호
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• pp.343-352
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• 2006

Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

• The Korean Journal of Physiology and Pharmacology
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• 제8권5호
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• pp.273-280
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• 2004

Nitric oxide (NO) has been suggested to act as a mediator of cytokine-induced effects of turn over of bone. Activation of the inducible nitric oxide synthase (iNOS) by inflammation has been related with apoptotic cell death in osteoblast. YS 49, a synthetic isoquinoline alkaloid, inhibits NO production in macrophages activated with cytokines. In the present study, we investigated the molecular mechanism of YS 49 to inhibit iNOS expression in ROS 17/2.8 cells, which were activated with combined treatment of inflammatory cytokines $(TNF-{\alpha},\;IFN-{\gamma})$ and lipopolysaccharide (LPS). Results indicated that YS 49 concentration-dependently reduced iNOS mRNA and protein expression, as evidenced by Northern and Western blot analysis, respectively. The underlying mechanism by which YS 49 suppressed iNOS expression was not to affect iNOS mRNA stability but to inhibit activation and translocation of $NF-_kB$ by preventing the degradation of its inhibitory protein $I_kB_{\alpha}$. As expected, YS 49 prevented NO-induced apoptotic cell death by sodium nitroprusside. Taken together, it is concluded that YS 49 inhibits iNOS expression by interfering with degradation of phosphorylated inhibitory $_kB_{\alpha}\;(p-I_kB_{\alpha})$. These actions may be beneficial for the treatment of inflammation of the joint, such as rheumatoid arthritis.

• 한국발생생물학회지:발생과생식
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• 제11권3호
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• pp.205-217
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• 2007

본 연구는 자궁근종 성장에 관한 분자생물학적인 기전의 이해를 위해 자궁근종 및 정상 자궁근세포의 초기 배양방법을 확립하기 위해 실시하였다. 이를 위해 최종적으로 두 가지 세포 배양 방법이 확립되었다. 그리고 안정적으로 연구(특히, 여성호르몬에 대한 반응 연구)에 사용할 수 있는 가장 적합한 세포 배양 방법이 모색되었다. 두 가지 세포 배양 조건 중 두 번째 방법(method 2)이 안정적으로 세포의 반응을 연구하는데 더 나은 방법으로 결론 내려졌고, 여성호르몬에 대한 반응이 더 좋은 것으로 밝혀졌다. 이 방법을 통해 배양된 세포에 $E_2$를 처리했을 때 정상 자궁근세포에 비해 근종세포에서 PR, IGF-1 and IGF-1 receptor mRNA의 발현이 더 높은 것으로 나타났다. 더욱이 주변에 있는 세포들보다는 조직에서 좀 더 큰 반응을 보였으며, 이는 $E_2$에 대한 세포의 반응에 세포외기질(extracellular matrix)과 세포 사이의 상호작용이 필요하다는 것을 의미한다. 결론적으로 이러한 근종세포 및 조직의 초기 배양 방법은 in vitro 상에서 종양 발생에 대한 기초연구를 위해 유용하게 사용될 수 있을 것으로 사료된다.

• 대한화장품학회지
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• 제31권2호
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• pp.141-146
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• 2005

불안정한 생리활성물질들은 외부 환경에 의해 빠르게 분해된다. 그러므로 이러한 물질들을 안정화시키기 위한 캡슐화 기술은 매우 중요하다. 비타민 $D_3$의 전구체인 7-디하이드로콜레스테롤(7-DHC)은 일반인의 표피 각화세포에서 열충격 단백질(Heat Shock Protein)의 발현을 단백질과 mRNA의 수준에서 증가시키는 것으로 알려졌다. 하지만 7-DHC의 국소용 피부 제제로의 이용은 낮은 용해도와 화학적 불안정성 때문에 이용이 제한되었다. 본 연구에서 7-DHC는 나노에멀젼(NE), 고형 지질 나노 입자 (SLN) 그리고 키토산이 코팅된 고형 지질 나노 입자(CASLN)에 봉입하였다. NE와 SLN은 지질의 용융점 이상의 온도에서 고압의 호모제나이져를 통과시켜 제조하였다. CASLN은 SLN 분산액에 키토산을 용액을 첨가하여 제조하였으며 양(+)의 제타전위를 나타내었다. NE, SLN, CASLN 속에서 7-DHC의 안정도를 각각의 온도조건에서 시간의 경과에 따라 확인하였다. 열분석과 X선 회절 분석은 지질의 결정화 정도를 확인하기 위해서 수행하였다. 그 결과, CASLN은 기존의 SLN보다 불안정한 7-DHC를 효과적으로 봉입함으로서 안정성을 개선시켰다.

• BMB Reports
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• 제35권6호
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• pp.623-628
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• 2002

The Th1 vs. Th2 balance is critical for the maintenance of immune homeostasis. Therefore, the genes that are selectively-regulated by the Th1 and Th2 cytokines are likely to play an important role in the Th1 and Th2 immune responses. In order to search for and identify the novel target genes that are differentially regulated by the Th1/Th2 cytokines, the human PBMC mRNAs differentially expressed upon the stimulation with IL-4 or IL-12, were screened by employing the differential display-polymerase chain reaction. Among a number of clones selected, DC21 was identified as a novel target gene that is regulated by IL-4 and IL-12. The DC21 gene expression was up-regulated either by IL-4 or IL-12, yet counter-regulated by co-treatment with IL-4 and IL-12. DC21 is a dendritic cell protein with an unknown function. The sequence analysis and conserved-domain search revealed that it has two AU-rich motifs in the 3'UTR, which is a target site for the regulation of mRNA stability by cytokines, and that it belongs to the N-acetyltransferase family. The induction of DC21 by IL-12 peaked around 8-12 h, and lasted until 24 h. LY294002 and SB203580 significantly suppressed the IL-12-induced DC21 gene expression, which implies that PI3K and p38/JNK are involved in the IL-12 signal transduction pathway that leads to the DC21 expression. Furthermore, tissue blot data indicated that DC21 is highly expressed in tissues with specialized-resident macrophages, such as the lung, liver, kidney, and placenta. Together, these data suggest a possible role for DC21 in the differentiation and maturation of dendritic cells regulated by IL-4 and IL-12.

• Journal of Veterinary Science
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• 제22권6호
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• pp.76.1-76.15
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• 2021

Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.