• 한국환경성돌연변이발암원학회지
• /
• 제25권2호
• /
• pp.76-81
• /
• 2005

25-Hydroxycholesterol (cholest-5-ene-3, 25-diol, 25-OHC) showed the cytotoxicity on HeLa human cervix and NCI-H460 human lung cancer cells, $0.5{\mu}M\;of\;50\%$ inhibitory concentration. We studied 25-OHC as the possibility of radiation sensitizer. The combination effect of 25-OHC and y-irradiation measured using flow cytometer with propidium iodide stained cells. The combined treatment of 25-OHC and $\gamma-irradiation$ did not show significant enhancing effects on Hela and NCI-H460 cells.

• Tuberculosis and Respiratory Diseases
• /
• 제76권3호
• /
• pp.114-119
• /
• 2014

Background: Epigallocatechin-3-gallate (EGCG), a major biologically active component of green tea, has anti-cancer activity in human and animal models. We investigated the schedule-dependent effect of EGCG and paclitaxel on growth of NCI-H460 non-small cell lung cancer cells. Methods: To investigate the combined effect of EGCG (E) and paclitaxel (P), combination indices (CIs) were calculated, and cell cycle analysis was performed. For the effect on cell apoptosis, western blot analysis was also performed. Results: CI analysis demonstrated that both concurrent and sequential E ${\rightarrow}$ P treatments had antagonistic effects (CIs >1.0), but sequential P ${\rightarrow}$ E had synergistic effects (CIs <1.0), on the growth inhibition of NCI-H460 cells. In the cell cycle analysis, although paclitaxel induced $G_2/M$ cell cycle arrest and increased the sub-G1 fraction, concurrent EGCG and paclitaxel treatments did not have any additive or synergistic effects compared with the paclitaxel treatment alone. However, western blot analysis demonstrated that sequential P ${\rightarrow}$ E treatment decreased the expression of Bcl-2 and procaspase-3 and increased poly(ADP-ribose) polymerase (PARP) cleavage; while minimal effects were seen with concurrent or sequential E ${\rightarrow}$ P treatments. Conclusion: Concurrent or sequential E ${\rightarrow}$ P treatment had opposite effects to P ${\rightarrow}$ E treatment, where P ${\rightarrow}$ E treatment showed a synergistic effect on growth inhibition of NCI-H460 cells by inducing apoptosis. Thus, the efficacy of EGCG and paclitaxel combination treatment seems to be schedule-dependent.

• 생명과학회지
• /
• 제23권2호
• /
• pp.167-174
• /
• 2013

Peroxiredoxin 6 (Prx 6)는 티올-특이적 항산화 단백질에 속하는 효소로서 산화적 스트레스로부터 세포를 보호할 뿐만 아니라 과산화물의 환원작용을 촉매한다. 본 연구에서는 인간 Prx 6의 promoter를 이용하여 항산화반응을 유발하는 추출물을 효과적으로 스크리닝하는 새로운 형질전환마우스를 개발하는 최종목적을 달성하기 위한 중간단계로서, hPrx 6/Luc 벡터를 개발하고, 이들 벡터의 안정적 발현과 성공적 반응성을 세포주를 이용하여 확인하고자 하였다. 이를 위해, 인간 Prx 6 promoter를 증폭하여 luciferase cDNA와 결합한 hPrx 6/Luc 벡터를 제조하였으며, 제조된 벡터를 제한효소 절단과 염기서열분석을 통해 확인하였다. hPrx 6/Luc 벡터는 NCI-H460 세포에 transfection한 후 인삼(KWG), 홍삼(KRG), 맥문동(LP), 홍문동(RLP)의 4가지 추출물을 처리하여 luciferase activity를 측정하였다. 그 결과, luciferase activity는 4가지 추출물에 의해 효과적으로 증가하였고, 특히 KRG과 LP를 처리한 그룹이 KWG과 RLP를 처리한 그룹보다 높았다. 또한, luciferase activity는 RLP 농도에 의존적으로 증가하였다. hPrx 6/Luc 벡터와 hPrx 6 mRNA반응의 차이를 비교하기 위해, 4가지 추출물을 처리한 후 hPrx 6 mRNA의 양을 RT-PCR로 분석하였다. 그러나, hPrx 6 mRNA의 양은 비록 고농도의 RLP 추출물에서는 약간의 증가가 관찰되었지만, 대조군에 비하여 4가지 추출물에서 유의적인 차이가 없었다. 한편, 4가지 추출물에 의한 superoxide dismutase (SOD) 활성의 변화는 비록 일부 차이는 있었지만 hPrx 6/Luc 벡터와 유사한 반응을 나타내었다. 따라서, 이러한 결과는 hPrx 6/Luc 벡터는 성공적으로 제조되었고, 세포내에서 안정적으로 발현하면서 항상화물질에 민감하고 정량적으로 반응할 수 있음을 제시하고 있다. 더불어, 이러한 세포주에서 확인결과를 바탕으로 형질전환마우스가 개발된다면, 항산화물질을 정량적으로 스크리닝하는 시스템으로 적용가능성이 매우 높음을 보여주고 있다.

• Tuberculosis and Respiratory Diseases
• /
• 제56권5호
• /
• pp.514-522
• /
• 2004

연구 배경 : 비스테로이드성 항염증제는 대장암의 화학 예방에 이용되고 있다. 지속적으로 비스테로이드성 항염증제를 복용한 결과 대장암 발생의 위험이 40-50% 감소하였다. Sulindac은 비스테로이드성 항염증제의 일종으로 대장암의 예방 효과가 있으며 암세포의 성장 억제와 세포 고사를 유도시킨다. 이에 저자들은 3가지 비소세포 폐암 세포주에서 sulindac의 영향을 알아보고자 하였다. 방 법 : A549(선암), NCI-H157(편평상피암), NCI-H460(대세포암) 세포주에 sulindac을 농도별로 투여하여, MTT assay로서 암세포의 생존율을, 유식세포 분석법과 핵산 염색으로 세포 고사의 비율을, 유산탈수소효소유리로서 세포 사멸의 정도를 시간대별로 측정하였다. 결 과 : Sulindac에 의해 농도와 시간에 의존적으로 비소세포 폐암 세포주에서 암세포의 생존율이 감소하였고, 유산 탈수소 효소 유리는 증가하였으며, 세포 고사 역시 농도, 시간에 의존적으로 증가하였다. 결 론 : Sulindac은 비소세포 폐암 세포주에서 농도, 시간에 의존적으로 암세포의 생존율 감소와 세포 고사증가를 유도하였다.

• 한국식품영양학회지
• /
• 제32권5호
• /
• pp.511-521
• /
• 2019

The purpose of this study was to investigate the inhibitory effect of enzyme activity and anti-proliferation of human cancer cell lines (HCT 116, NCI-H460 and MCF-7) of peanut skin depending on cultivars (Arachis hypogaea L. cv. K-Ol, cv. Sinpalkwang, cv. Daan, cv. Heuksaeng) and extraction solvent. Peanut skin was extracted with 80% ethanol, 80% methanol, 80% acetone, and distilled water, followed by analysis of the enzyme inhibitory activity and anticancer activity. Methanol extract of Daan cultivar most effectively inhibited ${\alpha}$-gluosidase (65.08%, 0.025 mg/mL), tyrosinase (82.49%, 2 mg/mL) and ACE (73.61%, 10 mg/mL). The inhibitory effect of peanut skin extracts on colon cancer cell (HCT-116), lung cancer cell (NCI-H460) and breast cancer cell (MCF-7) growth were investigate using MTT assay. The highest anti-proliferation of cancer cell line of peanut skin extracts was observed in the methanol extract of Daan cultivar. The cell viability on HCT 116, NCI-H460 and MCF-7 cell lines of methanol extracts from peanut skin of Daan cultivar was 48.13%, 41.03%, and 36.02% at $200{\mu}g/mL$, respectively. These results suggest that peanut skin extracts may mediate physiological activity, and provide valuable information for the use of peanut byproduct as a functional food material.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2005년도 제45회 학술심포지움 및 추계학술대회
• /
• pp.33-33
• /
• 2005

• Molecules and Cells
• /
• 제27권1호
• /
• pp.125-125
• /
• 2009

• Biomolecules & Therapeutics
• /
• 제13권4호
• /
• pp.220-226
• /
• 2005

The present study was performed to evaluate combination effect of 7$\beta$-hydroxycholesterol (7$\beta$-OHC) and $\gamma$-radiation in NCI-H460 human lung cancer cells. 7$\beta$-OHC in combination with $\gamma$-irradiation has an enhanced effect in decreasing clonogenic survival and increasing cellular DNA damage. Pretreatment of cells with 7$\beta$-OHC enhanced radiation-induced apoptosis. Apoptosis of the cells by combined treatment of 7$\beta$-OHC and $\gamma$-irradiation was associated with reactive oxygen species generation and loss of mitochondrial membrane potential, resulting in the activation of caspase 9 and caspase 3. The combined treatment also resulted in an increased G1 cell cycle distribution. These results indicate that 7$\beta$-OHC shows the additive effect of radiation sensitivity in human lung carcinoma cells in vitro.

• Tuberculosis and Respiratory Diseases
• /
• 제71권4호
• /
• pp.259-265
• /
• 2011

Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, gefitinib and erlotinib, are effective therapies for non-small cell lung cancer (NSCLC) patients whose tumors harbor somatic mutations in EGFR. The mutations are, however, only found in about 30% of Asian NSCLC patients and all patients ultimately develop resistance to these agents. Ionizing radiation has been shown to induce autophosphorylation of EGFR and activate its downstream signaling pathways. In the present study, we have tested whether the effect of gefitinib treatment can be enhanced after ionizing radiation. Methods: We compared the PC-9 and A549 cell line with its radiation-resistant derivatives after gefitinib treatment with cell proliferation and apoptosis assay. We also analyzed the effect of gefitinib after ionizing radiation in PC-9, A549, and NCI-H460 cells. Cell proliferation was determined by MTT assay and induction of apoptosis was evaluated by flow cytometry. Caspase 3 activation and PARP cleavage were evaluated by western blot analysis. Results: PC-9 cells having mutated EGFR and their radiation-resistant cells showed no significant difference in cell viability. However, radiation-resistant A549 cells were more sensitive to gefitinib than were their parental cells. This was attributable to an increased induction of apoptosis. Gefitinib-induced apoptosis increased significantly after radiation in cells with wild type EGFR including A549 and NCI-H460, but not in PC-9 cells with mutated EGFR. Caspase 3 activation and PARP cleavage accompanied these findings. Conclusion: The data suggest that gefitinib-induced apoptosis could increase after radiation in cells with wild type EGFR, but not in cells with mutated EGFR.

• 동의생리병리학회지
• /
• 제22권3호
• /
• pp.630-641
• /
• 2008

Gamisamgibopae-tang (GMSGBPT) is a traditional Korean medicine, which has been used for patients suffering from a lung disease in Oriental medicine. In the present study, we examined the biochemical mechanisms of apoptosis by GMSGBPT in NCI-H460 and A549 human non-small-cell lung cancer cell lines. It was found that GMSGBPT could inhibit the cell proliferation of A549 cells in a concentration-dependent manner, however GMSGBPT did not affect the cell proliferation of NCI-H460 cells. Apoptotic cell death in A549 cells were detected using DAPI staining and annexin V fluorescein methods. The induction of apoptotic cell death by GMSGBPT was connected with a down-regulation of anti-apoptotic Bcl-2 and Bcl-xL expression, and proteolytic activation of caspase-3 and caspase-9 in A549 cells. However, GMSGBPT did not affect the levels of pro-apoptotic Bax and Bad expression, and activity of caspase-8. GMSGBPT treatment also concomitant degradation and/or inhibition of poly (ADP-ribose) polymerase (PARP), ${\beta}$-catenin, phospholipase C-1 (PLC${\gamma}$1) and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Taken together, these findings suggest that GMSGBPT may be a potential chemotherapeutic agent for the control of human non-small-cell lung cancer cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of GMSGBPT.