• 대한의생명과학회지
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• 제10권3호
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• pp.263-268
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• 2004

This study was carried out to evaluate the cytotoxic effect of phenolic compound on normal NIH 3T3 fibrolasts. The colorimetric assay for phenol compound, syringic acid was performed by MTT assay or XTT assay. MTT or XTT assays are known as a very sensitive method in measuring the cytotoxic effect of chemical agents in vitro. In the present study, syringic acid on normal Nlli 3T3 fibroblasts did not show any cytotoxicity for MTT assay or XTT assay compared with control after cells were treated with various concentrations of syringic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 3,340.9 μM and 2,462.4 μM of syringic acid, respectively. From the above the results, it is suggested that phenolic compound of syringic acid did not have any cytotoxicity on normal NIH 3T3 fibroblasts.

• BMB Reports
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• 제49권2호
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• pp.99-104
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• 2016

The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

• 대한본초학회지
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• 제21권1호
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• pp.101-107
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• 2006

Objectives : It is well known that hexavalent chromium has toxic effect on normal cells. Recently, toxic effect of hexavalent chromium is diminished by the some extracts derived from herbs or plants. But, the toxic or protective mechanism of hexavalent chromium is well unknown. This study was performed to examine the protective effect of poncirin against $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Methods : The protective effect of the cytotoxicity induced by $Na_2Cr_2O_7$ was measured by the cell viability after NIH3T3 fibroblasts were cultured with or without $Na_2Cr_2O_7$ for 48 hours. Antitoxic effects of poncirin on the cytotoxicity induced by $Na_2Cr_2O_7$ were examined by colorimetric assays such as MTT or XTT assay. Results : $Na_2Cr_2O_7$ decreased cell viability by the decreased absorbance in MTT or XTT assay, but, the poncirin increased cell viability which was decreased by $Na_2Cr_2O_7$-induced cytotoxicity on NIH3T3 fibroblasts. Conclusion : These results suggest that $Na_2Cr_2O_7$ showed cytotoxicity effect on NIH3T3 fibroblasts by the decrease of cell viavility, and poncirin was effective in the protection of $Na_2Cr_2O_7$-induced cytotoxicity in these cultures.

• 대한화장품학회지
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• 제24권1호
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• pp.74-86
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• 1998

자외선에 의한 유전독성의 감소에 미치는 홍삼추출물의 영향을 배양 NIH3T3 세포계에서 분석하였다. 자외선을 조사한 후 정상 배지에서 배양한 시간간격에 따라 세포의 생존률은 증가하였는데 홍삼추출물이 함유된 배지에서 배양한 경우는 약 15%정도 증가한 생존률을 보였다. 자외선을 조사한 후 감소된 DNA복제가 정상배지 배양시간에 따라 증가하는 정도도 홍삼추출물을 후처리할 경우 현저한 증가를 보였다. 자외선 상해를 회복하기 위한 절제회복능은 홍삼추출물을 처리할 경우 유의미한 증가를 보였다. 이러한 절제회복과정 중 효소에 의한 절제단계가 홍삼추출물 처리에 의해 활성화됨을 단사절단 분석을 통하여 규명하였다. 이상의 결과는 홍삼추출물이 자외선 상해의 절제회복에 유의미한 증가를 보이며 따라서 유전독성을 감소시키는 항노화제로써 사용할 수 있음을 시사한다.

• 산업기술연구
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• 제25권B호
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• pp.241-251
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• 2005

Exopolysaccharide (CBP) from submerged culture broth of Ganoderma lucidum mycelium and the water soluble (BWS) and water insoluble (BWI) fractions of CBP were prepared by gel filtration. Antitumor activity and effects on proliferation and differenciation of human cancer cells and mouse NIH 3T3 cells were studied. Cytotoxicity test of CBP, BWS and BWI fractions on human cancer cell lines was performed by using sulforhodamine B (SRB) assay. A549 (lung carcinoma), Colo320 DM and HSR (colon carcinoma), and NIH 3T3 cells were used. BWI fraction showed the strongest cytotoxicity (maximum 20% survival) to all human cells tested. However it did not induced apoptosis. Interestingly BWI fraction did not exert cytotoxic effect on NIH 3T3 cells at low concentration of cells ($5{\times}10^4$) but strong toxic effect at high concentration of cells($5{\times}10^5$) which showed transformed morphology. These results suggest that BWI may have cancer cell specific anticancer activity. However, BWI fraction did not effect the amount of pRb and c-myc protein, which implied that BWI fraction did not act at the early stage of signal transduction pathway. CBP fraction induced differenciation of human leukemic cell line, HL-60 cells suggesting the carcinogenesis prevention of normal cell and possible induction of normalization for cancer cell.

• Journal of Microbiology and Biotechnology
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• 제25권12호
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• pp.1989-1996
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• 2015

Ice-binding proteins (IBPs) can inhibit ice recrystallization (IR), a major cause of cell death during cryopreservation. IBPs are hypothesized to improve cell viability after cryopreservation by alleviating the cryoinjury caused by IR. In our previous studies, we showed that supplementation of the freezing medium with the recombinant IBP of the Arctic yeast Glaciozyma sp. (designated as LeIBP) could reduce post-thaw hemolysis of human red blood cells and increase the survival of cryopreserved diatoms. Here, we showed that LeIBP could improve the viability of cryopreserved mammalian cells. Human cervical cancer cells (HeLa), mouse fibroblasts (NIH/3T3), human preosteoblasts (MC3T3-E1), Chinese hamster ovary cells (CHO-K1), and human keratinocytes (HaCaT) were evaluated. These mammalian cells were frozen in dimethyl sulfoxide (DMSO)/fetal bovine serum (FBS) solution with or without 0.1 mg/ml LeIBP at a cooling rate of -1℃/min in a -80℃ freezer overnight. The minimum effective concentration (0.1 mg/ml) of LeIBP was determined, based on the viability of HeLa cells after treatment with LeIBP during cryopreservation and the IR inhibition assay results. The post-thaw viability of mammalian cells was examined. In all cases, cell viability was significantly enhanced by more than 10% by LeIBP supplementation in 5% DMSO/5% FBS: viability increased by 20% for HeLa cells, 28% for NIH/3T3 cells, 21% for MC3T3-E1, 10% for CHO-K1, and 20% for HaCaT. Furthermore, addition of LeIBP reduced the concentrations of toxic DMSO and FBS down to 5%. Therefore, we demonstrated that LeIBP can increase the viability of cryopreserved mammalian cells by inhibiting IR.

• 한국해양바이오학회지
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• 제7권1호
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• pp.28-34
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• 2015

We examined cell regeneration efficiency of brown marine algae living in Jeju coast for search of a novel therapeutic device with cutaneous wound healing materials. The five algae were collected and compared with epidermal growth factor (EGF) as a positive control in the assays of cell proliferation and cell migration of NIH3T3 fibroblast cells. Among the 80% methanol extracts of these brown algae, the two algal extracts from Ishige foliacea and Colpomenia bullosa showed the proliferative effects of the cells similar to the effect of EGF. Besides it was found that Colpomenia bullosa extract significantly enhanced cell migration of NIH3T3 cell. In the study, therefore, we confirmed that the Colpomenia bullosa extract improved proliferation of NIH3T3 cell and a potential candidate for cultaneous wound healing.

• Bulletin of the Korean Chemical Society
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• 제34권4호
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• pp.1165-1169
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• 2013

To elucidate the role of PKG isoforms in transcriptional control of cyclin D1, we employed a series of expression vectors of PKG $1{\alpha}$ and PKG $1{\beta}$ which encode HA-tagged wild type and constitutively active (SD and ${\Delta}N$) mutants. Our present study demonstrates that both the constitutively active mutants of PKG $1{\beta}$ downregulate the transcription of cyclin D1 when transiently transfected in NIH3T3 cells, whereas PKG $1{\alpha}$ mutants show weak inhibition. We further studied the transcriptional regulators of cyclin D1, such as, c-fos, NF-${\kappa}B$, and CRE by using the luciferase reporter assay. Constitutively active mutants of PKG $1{\beta}$ showed marked transcriptional downregulation of c-fos in NIH3T3 cells, whereas PKG $1{\alpha}$ downregulated c-fos to a lesser extent. We also found that the constitutively active mutants of PKG negatively regulated the activation of NF-${\kappa}B$ and CRE, suggesting their involvement in the regulation of cyclin D1.

• KSBB Journal
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• 제24권4호
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• pp.397-402
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• 2009

본 연구에서는 몇 가지 생물공학 기반 기술로서 이미 한방 화장품의 소재로 사용되고 있는 쇠비름 에탄올 추출액의 tyrosinase 저해, collagen 합성, 항염증 기대 효과, 항노화 대한 효능을 알아보기 위하여 B16F10 mouse melanoma cell, NIH3T3 mouse embryonic fibroblast, MCF-7 human breast adenocarcinoma cell에 쇠비름 추출액을 처리하여 실험하였다. 실험 결과, 쇠비름 에탄을 추출액 (0.5mg/mL)은 tyrosinase 발현을 억제하여 멜라닌 합성을 억제하며, 농도 의존적으로 NIH3T3 mouse fibroblast 세포의 collagen 합성을 촉진시켰다. 또한 2.0 mg/mL 농도의 쇠비름 추출액은 목단피보다 더욱 효과적으로 cytokine (TNF-$\alpha$)에 의한 NF-${\kappa}B$ 활성을 억제시켰다. Doxorubicin에 의한 과노화에서도 효과적인 항노화 작용을 하는 것으로 나타났다. Tyrosinase 합성을 억제하므로 미백에 대한 효과와 세포의 생장을 도와 collagen 합성을 촉진함으로서 노화방지에 효과적인 것으로 사료되며, TNF-$\alpha$에 의한 NF-${\kappa}B$의 활성화를 저해함으로서 염증반응 억제 효과도 기대 할 수 있다.

• 한국환경농학회지
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• 제20권3호
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• pp.155-161
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• 2001

Paraquat와 bentazone이 횐쥐의 간조직과 NIH 3T3 섬유모세포에 미치는 독성과 그 독성에 대한 3-MC의 보상효과를 조사하기 위하여 NIH 373 섬유모세포에 적응한 후 경시적으로 MTT분석을 이용하여 세포독성을 측정하고 Sprague Dawley계 웅성 횐쥐에 paraquat와 bentazone단독 및 paraquat 및 bentazone과 3-MC를 병용투여한 후 경시적으로 관찰한 결과 paraquat와 bentazone은 NIH 3T3 섬유모세포에 대하여 $IC_{50}$값이 각각 1668.97 ${\mu}M$, 1506.97 ${\mu}M$으로 Borenfreund의 독성평가기준에 의하면 저독성이었다. Paraquat와 bentazone 단독투여군의 H&E 염색에서 3시간째에는 문맥 주위 세포들이 변성을 일으키고 별모양 세포들이 증가하였으나 12시간째에는 간소엽 전체의 세포들이 변성을 일으켰으며 48시간째에는 더욱 심한 변성이 일어났다. 특히 bentazone 투여 후 48시간째에는 핵농축현상이 뚜렷하였다. Best carmine 염색에서 glycogen 과립을 함유하는 간세포들도 3시간째에는 문맥주위의 세포들이, 12시간째에는 간소엽 전체의 간세포들이, 48시간째에는 전체의 간세포들이 함유하는 glycogen 과립량이 현저히 증가하였다. 3-MC를 paraquat와 bentazone과 동시에 투여한 군에서 3시간째와 12시간째에는 단독투여군과 유사하였으나 48시간째에는 bentazone과 3-MC 동시투여군의 문맥주위의 간세포들이 재생되는 경향이었으며 paraquat와 3-MC를 동시에 투여한 군에서는 중심정맥 주위의 세포들만이 glycogen과립을 함유하고 있어 단독 투여군과 뚜렷한 차이를 관찰할 수 있었다. 이 결론에서 3-MC는 paraquat와 bentazone에 의한 간세포의 독성을 경감시킬 수 있는 물질임을 알 수 있었다.