• Tuberculosis and Respiratory Diseases
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• 제50권1호
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• pp.67-75
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• 2001

배경 : 대표적인 종양억제유전자인 p53 및 p16은 세포주기 조절 및 자연괴멸 유도에서 서로 다른 역할을 하고 있으며 폐암에서는 동시에 비활성화 되어 있는 경우가 흔하다. 이 종양억제유전자들 동시에 이입시 서로 상호작용을 통해 효과가 증진되리라 기대되고 있다. 방법 : 본 연구에서는 p53 및 p16을 아데노바이러스 벡터를 이용하여 폐암세포주에 동시에 이입하여 그 상호작용을 관찰하였다. 복합투여시 세포성장곡선을 분석하여 isobologra을 이용한 상호작용을 검증하고 세포주기의 변화 및 체외종양형성능의 변화도 관찰하였다. 결과 : Isobologram을 이용한 분석에서 adeno-virus-p53와 adenovirus-p16의 복합투여는 NCI H358에서는 상승적인 성장억제를, NCI H23에서는 부가적인 성장 억제를 보였다. 유세포분석기를 이용한 세포주기의 분석 및 체외종양형성능 검사에서도 p53 및 p16의 복합투여시 단독 투여보다 강한 억제효과를 보였다. 결론 : 이러한 상승적인 p53 및 p16의 상호작용은 이 복합요법이 새로운 유전자치료법으로 발전할 수 있는 가능성을 보였다.

• Radiation Oncology Journal
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• 제7권2호
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• pp.149-156
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• 1989

HeLa 세포의 spheroid를 배 양시켜 cis-platinum과 carboplatin으로 처리한 후 그 반응을 세포의 생존분획으로 분석하였다. 체외실험 model인 spheroid를 사용하여 platinum유사체의 약효와 방사선 감수성을 평가하고 약제에 대한 단층 세포와 spheroid의 감수성 차이를 세포-생존곡선에서 규명하기 위하여 본 실험을 시행하였다. Cis-platinum 농도-곡선에서 spheroid의 $Cq=3.4{\mu}M$이고 $Co=1.2{\mu}M$이었다. 이 것은 단층세포에 비하여 Co는 큰 변화가 없으나 Cq가 증가되어 cis-platinum이 저산소층 세포보다 활동적으로 분화하는 표면세포에 주로 작용하였으며, 반대로 carboplatin의 효과는 spheroid에 대한 $(Co=15.0{\mu}M)$ 감수성이 단층세포$(Co=32.5{\mu}M)$에 비하여 크게 증가되어, spheroid의 심층 세포에 주로 작용하였다. 방사선과 carboplation의 병용효과를 세포 생존분획이 0.01 수준에서 isobologram으로 분석한 결과 상호작용으로 supra-additive 효과를 나타내었다. 따라서, carboplatin은 cis-platinum에 비하여 신장과 위장에 대한 독성작용이 적고, 방사선과 병용함으로써 향후 더욱 효과적 인 종양 치료에 중요한 역할을 할 것으로 기대한다.

• 대한미생물학회지
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• 제21권3호
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• pp.375-380
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• 1986

Thirty-one strains of Pseudomonas aeruginosa were submitted to the synergistic activity test of amikacin(AK) and gentamicin(GM) combined with moxalactam(MX), ceftizoxime(CTZ) or cefoperazone(CFZ). The minimal inhibitory concentrations(MICs) of each drug and drugs combined in various ratios were measured by checkerboard dilution method. The synergism was determined through analysing the MIC distribution curve on isobologram and calculating the fractional inhibitory concentration index(FICI). MICs of GM, AK, MX, CFZ and CTZ against the 31 tested strains were distributed from $12.5{\mu}g/ml$ to $800{\mu}g/ml$, from $0.8{\mu}g/ml$ to $25{\mu}g/ml$, from $3.1{\mu}g/ml$ to $50{\mu}g/ml$, from $3.1{\mu}g/ml$ to $400{\mu}g/ml$, and from $12.5{\mu}g/ml$ to $100{\mu}g/ml$, respectively. The rate synergism of each drug combination by means of FICl was 45.5% in GM-MX, 36.4% in GM-CFZ, 63.6% in GM-CTZ, 48.6% in AK-MX, 35.3% in AK-CFZ, and 35.7% in AK-CTZ combination. Thus, it is suggested that Pseudomonas aeruginosa may effectively be inhibited by various aminoglycoside and cephalosporin combinations.

• Natural Product Sciences
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• 제25권4호
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• pp.311-316
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• 2019

Artocarpus heterophyllus has been used as traditional medicine. This plant is one of the sources of flavonoid. Flavonoid compounds possessed a wide range of biological properties including anticancer. This study was performed to investigate the cytotoxic effect of flavonoids from A. heterophyllus on H460 and MCF-7 cell lines. The interaction of flavonoids and cisplatin against tested cancer cells was also evaluated. MTT assay was used to determine the cytotoxic effect of flavonoid. Isobologram analysis was selected to evaluate the synergistic effect between flavonoid and cisplatin, their interaction was then confirmed using AO/PI staining method. Amongst of flavonoid compounds, artocarpin exhibited strong cytotoxic effect on both MCF-7 and H460 cell lines with IC₅₀ values of 12.53 ㎍/mL (28.73 μM) and 9.77 ㎍/mL (22.40 μM), respectively. This compound enhanced anticancer activity of cisplatin against H460 and MCF-7. The combination produced a synergistic effect on H460 and MCF-7 cell lines with a combination index (CI) values of 0.2 and 0.18, respectively. The AO/PI stained demonstrated that the combination of artocarpin and cisplatin caused morphological changes that indicated apoptosis. Moreover, artocarpanone also significantly increased cytotoxic effect of cisplatin compared to its single concentration with CI below than 1. This result suggested the potency of flavonoid named artocarpin to enhance the anticancer activity of cisplatin on H460 and MCF-7 cell lines.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.473-481
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• 2020

Axl receptor tyrosine kinase has been implicated in cancer progression, invasion, and metastasis in various cancer types. Axl overexpression has been observed in many cancers, and selective inhibitors of Axl, including R428, may be promising therapeutic agents for several human cancers, such as breast, lung, and pancreatic cancers. Here, we examined the cell growth inhibition mediated by R428 and auranofin individually as well as in combination in the human breast cancer cell lines MCF-7 and MDA-MB-231 to identify new advanced combination treatments for human breast cancer. Our data showed that combination therapy with R428 and auranofin markedly inhibited cancer cell proliferation. Isobologram analyses of these cells indicated a clear synergism between R428 and auranofin with a combination index value of 0.73. The combination treatment promoted apoptosis as indicated by caspase 3 activation and poly (ADP-ribose) polymerase cleavage. Cancer cell migration was also significantly inhibited by this combination treatment. Moreover, we found that combination therapy significantly increased the expression level of Bax, a mitochondrial proapoptotic factor, but decreased that of the X-linked inhibitor of apoptosis protein. Furthermore, the suppression of cell viability and induction of Bax expression by the combination treatment were recovered by treatment with N-acetylcysteine. In conclusion, our data demonstrated that combined treatment with R428 and auranofin synergistically induced apoptosis in human breast cancer cells and may thus serve as a novel and valuable approach for cancer therapy.

• Fisheries and Aquatic Sciences
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• 제26권2호
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• pp.133-144
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• 2023

Auranofin is a US Food and Drug Administration (FDA)-approved anti-arthritis medication that functions as a thioredoxin reductase inhibitor. Spermidine, a polyamine present in marine algae, can exert various physiological functions. Herein, we examined the synergistic anticancer activity of auranofin and spermidine in hepatocellular carcinoma (HCC). Combined treatment with auranofin and spermidine suppressed cell viability more efficiently than either treatment alone in HCC Hep3B cells. The isobologram plotted by calculating the half maximal inhibitory concentration (IC₅₀) values of each drug indicated that the two drugs exhibited a synergistic effect. Based on the analysis of annexin V and cell cycle distribution, auranofin and spermidine markedly induced apoptosis in Hep3B cells. Moreover, auranofin and spermidine increased mitochondria-mediated apoptosis by promoting mitochondrial membrane potential (Δψm) loss. Auranofin and spermidine significantly increased reactive oxygen species (ROS) production in Hep3B cells, and the blocking ROS suppressed apoptosis induced by spermidine and auranofin. In addition, auranofin and spermidine reduced the expression of phosphorylated phosphatidylinositol-3 kinase (PI3K) and protein kinase B (Akt), and PI3K inhibitor accelerated auranofin- and spermidine-induced apoptosis. Using ROS scavenger and PI3K inhibitor, we revealed that ROS acts upstream of auranofin- and spermidine-induced apoptosis. Collectively, our study suggests that combination treatment with auranofin and spermidine could afford synergistic anticancer activity via ROS overproduction and reduced PI3K/Akt signaling pathway.

• Biomolecules & Therapeutics
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• 제29권2호
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• pp.234-247
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• 2021

We used a heterozygous gene deletion library of fission yeasts comprising all essential and non-essential genes for a microarray screening of target genes of the antifungal terbinafine, which inhibits ergosterol synthesis via the Erg1 enzyme. We identified 14 heterozygous strains corresponding to 10 non-essential [7 ribosomal-protein (RP) coding genes, spt7, spt20, and elp2] and 4 essential genes (tif302, rpl2501, rpl31, and erg1). Expectedly, their erg1 mRNA and protein levels had decreased compared to the control strain SP286. When we studied the action mechanism of the non-essential target genes using cognate haploid deletion strains, knockout of SAGA-subunit genes caused a down-regulation in erg1 transcription compared to the control strain ED668. However, knockout of RP genes conferred no susceptibility to ergosterol-targeting antifungals. Surprisingly, the RP genes participated in the erg1 transcription as components of repressor complexes as observed in a comparison analysis of the experimental ratio of erg1 mRNA. To understand the action mechanism of the interaction between the drug and the novel essential target genes, we performed isobologram assays with terbinafine and econazole (or cycloheximide). Terbinafine susceptibility of the tif302 heterozygous strain was attributed to both decreased erg1 mRNA levels and inhibition of translation. Moreover, Tif302 was required for efficacy of both terbinafine and cycloheximide. Based on a molecular modeling analysis, terbinafine could directly bind to Tif302 in yeasts, suggesting Tif302 as a potential off-target of terbinafine. In conclusion, this genome-wide screening system can be harnessed for the identification and characterization of target genes under any condition of interest.

• 대한골관절종양학회지
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• 제4권1호
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• pp.13-21
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• 1998

Taxol이 악성 골종양 세포에 어느 정도의 세포독성이 있는지를 평가하기 위해 한국세포주 은행에서 분양 받은 G-292, SaOS-2 및 HT-1080의 3가지 악성 골종양 세포들을 대상으로 기존의 항암제인 methotrexate, adriamycin, ifosfamide, cisplatinum과 함께 각각 투여하여 MTT분석법으로 정량 및 비교 분석하였으며, adriamycin과 taxol을 병용 투여하여 항암제의 상호작용을 isobologram 분석법으로 조사하여 다음과 같은 결과를 얻었다. 1. Taxol의 악성 골종양 세포 주들에 대한 $IC_{50}$는 G-292에서는 $2.7{\times}10^{-2}{\mu}g/ml$, $SaOS^{-2}$에서는 $1.0{\times}10^{-2}{\mu}g/ml$, $HT{\times}1080$에서는 $1.1{\times}10^{-3}{\mu}g/ml$이었다. 2. Taxol은 악성 골종양 세포주들에 대해 기존의 항암제들 보다 강한 세포독성을 보였으며, 기존 항암제의 세포 독성의 강도는 adriamycin이 제일 높은 역가를 보였으며 그 외 methotrexate, cisplatinum, ifosfamide순이었다. 3. Taxol과 adriamycin을 병용 투여하여 상호작용을 관찰한 결과 G-292와 SaOS-2 세포 주에서 상승효과가 관찰되었으며, HT-1080에서는 상승효과가 관찰되지 않았다.