레스베라트롤에 의한 인간 암세포주, A549와 SKOV3의 p53의존적 Apoptosis 유발

이슬기 ( Seul Gi Lee ),남주옥 ( Ju Ock Nam ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.2

Resveratrol은 포도, 오디, 땅콩과 같은 많은 과일과 채소에 존재하는 폴리페놀 화합물로써 다양한 생물학적 효과를 가진다고 보고되어있다. 그러나, resveratrol이 A549 폐암세포에서 유도하는 apoptosis에 관여하는 분자적 기전 및 anoikis에 관해서는 명백하게 밝혀지지 않았다. 본 연구에서, 우리는 정상적인 p53 유전자를 갖는 A549 세포에서 resveratrol의 효과를 조사하고, p53 유전자가 결실된 SKOV3 난소암세포와 그 효과를 비교했다. Resveratrol은 확실하게 SKOV3 세포에 비해 농도, 시간의존적으로 A549 세포의 생존과 증식을 억제했다. 또한 resveratrol은 A549 세포의 apoptosis를 유도했지만 세포의 anoikis 저항성에는 영향을 미치지 않았다. 더불어, p53 유전자 기능이 불완전 소실된(knockdown) A549 세포의 생존과 증식은 resveratrol에 의해 변하지 않았다. 그러므로, 본 연구의 결과는 resveratrol의 항암 효과가 기능을 하는 p53 유전자의 존재에 의존한다는 것을 보여준다. 결론적으로, 우리는 resveratrol이 p53 유전자에 의존적으로 A549 세포에 대해 항암효과를 가진다는 것을 입증했다. Resveratrol, a polyphenolic compound present in many fruits and vegetables such as grapes, mulberries, and peanuts, has been reported to have various biological effects. However, the molecular mechanisms underlying resveratrol-induced apoptosis in A549 ovarian cancer cells are not well understood. In this study, we investigated the effect of resveratrol on A549 lung cancer cells (expressing wild-type p53) and compared it with that observed for SKOV3 ovarian cancer cells (expressing null-type p53). Resveratrol significantly inhibited the viability and proliferation of A549 cells in a concentration- and time-dependent manner, compared with its effects on SKOV3 cells. It also induced A549 cell apoptosis, but did not affect anoikis resistance. Furthermore, the viability and proliferation of p53-knockdown A549 cells were unaffected by the presence of resveratrol. Therefore, we demonstrate that the anticancer effect of resveratrol against A549 lung cancer cells is dependent on the presence of functional p53.

Transglutaminase-2 induces N-cadherin expression in TGF-β1-induced epithelial mesenchymal transition via c-Jun-N-terminal kinase activation by protein phosphatase 2A down-regulation

Park, M.K.,You, H.J.,Lee, H.J.,Kang, J.H.,Oh, S.H.,Kim, S.Y.,Lee, C.H. Pergamon Press 2013 European journal of cancer Vol.49 No.7

Epithelial-mesenchymal-transition (EMT) is a key event for tumour cells to initiate metastasis leading to switching of E-cadherin to N-cadherin. Transglutaminase-2 (Tgase-2) expression is increased in TGF-β1-induced EMT in A549 lung cancer cells or other lung cancer cells. The role and underlying mechanism of Tgase-2 in N-cadherin switching of TGF-β1-induced EMT are not known. The involvement and mechanisms of Tgase-2 were investigated in A549 cells using chemical inhibitors, gene silencing and over-expression. TGF-β1-induced EMT was suppressed by cystamine or gene silencing of Tgase-2. Suppression of Tgase-2 or the c-Jun-N-terminal kinase (JNK) inhibitor, SP600125, significantly reduced and over-expression of Tgase-2 increased the expression of N-cadherin. The relationship between Tgase-2 and JNK in the TGF-β1-induced EMT of A549 cells was examined using Tgase-2 over-expressed A549 cells (A549<SUB>TG2</SUB>) and Tgase-2 silenced A549 cells (A549<SUB>shTG2</SUB>). JNK activation was significantly increased in A549<SUB>TG2</SUB> cells and decreased in A549<SUB>shTG2</SUB> cells. In contrast, PP2A expression was decreased in A549<SUB>TG2</SUB> and A549 cells and increased in A549<SUB>shTG2</SUB> cells. The involvement of Tgase-2 in N-cadherin expression was also confirmed in an in vivo lung cancer orthotopic model by injection of A549<SUB>WT</SUB> and A549<SUB>shTG2</SUB> cells into SCID mice. Tgase-2 expressing A549<SUB>WT</SUB> cells-injected mice group showed increased expressions of N-cadherin and JNK activation, but decreased expression of PP2A in lung cancer tissue comparing with the A549<SUB>shTG2</SUB> cells-injected group. These results suggested that Tgase-2 induces N-cadherin expression of TGF-β1-induced EMT via JNK activation by PP2A down-regulation, and Tgase-2/PP2A/JNK might be a novel axis that affects N-cadherin switching in the EMT of A549 lung cancer cells.

인체폐암세포 NCI-H460 및 A549의 apoptosis 유발에 미치는 삼기보배탕의 영향

허만규,허태율,김기탁,변미권,김진영,심성흠,김광록,감철우,박동일,Heo, Man-Kyu,Heo, Tae-Yool,Kim, Ki-Tak,Byun, Mi-Kwon,Kim, Jin-Young,Sim, Sung-Heum,Kim, Koang-Lock,Kam, Cheol-Woo,Park, Dong-Il 대한한방내과학회 2007 大韓韓方內科學會誌 Vol.28 No.3

Objectives : This study was designed to investigate the antiproliferative activity of the water extract of Samgibopae-tang (SGBPT) in NCI-H460 and A549 non-small-cell lung cancer cell lines Methods : In this study, we measured the subsistence, form of NCI-H460 and A549 non-small-cell lung cancer cell by hemocytometer and DAPI staining. In each cell, we analyzed DNA fragmentation. reverse transcription-polymerase chain reaction and measured activity of caspase-3, caspase-8 and caspase-9. Results and Conclusions : We found that exposure of A549 cells to SGBPT resulted in growth inhibition in a dose-dependent manner. butSGBPT did not affect the growth of NCI-H460 cells. The antiproliferative effect by SGBPT treatment in A549 cells was associated with morphological changes. SGBPT treatment partially induced the expression of DR5 cells and the expression of Faswas markedly increased in both transcriptional and translational levels in A549 cells. SGBPT treatment partially induced the expression of Bcl-2, Bcl-XL and the expression of Bid was markedly decreased in translational levels in A549 cells. However, SGBPT treatment did not affect the expression of IAP family in A549 orNCI-H460 cells. SGBPT treatment partially induced the expression of caspase-3, caspase-8, caspase-9 activity which markedly increased in a dose-dependent manners in A549 cells. The fragmental development of PARP and ${\beta}$-catenin protein was observed in A549 cells by SGBPT treatment. SGBPT treatment induced the expression of PLC-${\gamma}1$ protein which decreased in A549 cells. SGBPT treatment partially induced the expression of DFF45/ICAD which markedly increased in a dose-dependent manner in A549 cells. Taken together. these findings suggested that SGBPT-induced inhibition of human lung carcinoma did not affect NCI-H460 cell growth. However, SGBPT-induced inhibition of human lung carcinoma A549 cell growth was associated with the induction of death receptor and mitochondrial pathway. The results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of SGBPT.

Anti-proliferative Effects of β-ionone on Human Lung Cancer A-549 Cells

Sun Min Lee(이선민),Young Sook Kim(김영숙),Wook Jin Jang(장욱진),Abdur Md. Rakib(압두르 라키브),Tae Woo Oh(오태우),Boh Hyun Kim(김보현),So Young Kim(김소영),Jeong Ok Kim(김정옥),Yeong Lae Ha(하영래) 한국생명과학회 2013 생명과학회지 Vol.23 No.11

β-Ionone의 인체 비소폐암세포 A-549 cells (human non-small lung cancer A-549 cell)에 대한 anti-proliferative effect에 관한 연구를 수행하였다. A-549 cell에 다양한 농도의 β-ionone (1, 5, 10, and 15 μM)을 2, 4, 6일 간 처리 하고, 배양 2일에 A-549 cell의 생육억제와 관련되는 biochemical marker를 측정하였다. β-Ionone은 A-549 cell 생육을 dose와 time 의존적으로 저해하였다. β-Ionone 배양 2일의 IC50은 5.0 μg/ml이었다. β-Ionone은 농도 의존적으로 apoptosis를 유도하였다. β-Ionone은 p53, p21 및 Bax protein 수준을 증가시켰으나, Bcl-2 protein 발현은 억제시켰다. β-ionone은 cytosol cytochrome c 함량을 증가시켰고, caspase-9과 caspase-3 효소 활성증가를 유도하였다. 또한, β-ionone은 cPLA₂와 COX-2 protein level을 감소시켰다. 이와 같은 결과는 β-ionone의 A-549 cell에 대한 생육억제효과는 Bax와 Bcl-2 gene 발현을 reciprocal regulation하여 유도한 apoptosis와 cPLA₂ 및 COX-2 protein 발현 억제에 기인함을 의미한다. The anti-proliferative activity of β-ionone was investigated on human non-small lung cancer A-549 cells (designated A-549 cells). A-549 cells were treated with various concentrations of β-ionone (1, 5, 10, and 15 μM) for two, four, and six days. Biochemical markers related to the growth inhibition of A-549 cells by β-ionone were measured at the second day of incubation. β-Ionone inhibited the growth of A-549 cells by dose-and time-dependent manners, resulting in an IC50 of 5.0 μg/ml at the second day of incubation. β-Ionone induced apoptosis by a dose-dependent manner. β-Ionone increased levels of p53, p21, and Bax proteins, but suppressed expression of the Bcl-2 protein. Similarly, β-ionone enhanced cytochrome c release from the mitochondria to the cytosol, and induced activation of caspase-9 and -3. Additionally, β-ion-one reduced cPLA₂ and COX-2 protein levels. These results suggest that the β-ionone inhibits the proliferation of A-549 cells through reciprocal regulation of Bax and Bcl-2 gene expression and suppression of cPLA₂ and COX-2 protein expressions.

인체 폐암 세포주 A549에서 Euonymus porphyreus 추출물의 항산화 및 항암활성 분석

진수정(Soojung Jin),오유나(You Na Oh),손유리(Yu Ri Son),배수빈(Soobin Bae),박정하(Jung-ha Park),김병우(Byung Woo Kim),권현주(Hyun Ju Kwon) 한국생명과학회 2021 생명과학회지 Vol.31 No.2

Euonymus porphyreus는 노박덩굴과에 속하는 식물로 동아시아 지역에 널리 분포하며, 식물학상의 특징에 대한 보고는 있으나 항산화능과 항암활성 등에 관한 연구는 아직까지 밝혀진 바가 없다. 이에 본 연구에서는 인체 폐암세포인 A549를 사용하여 E. porphyreus 에탄올 추출물(EEEP)의 항산화 및 항암활성과 그 분자적 기전에 관하여 연구하였다. 먼저 EEEP의 총 폴리페놀 화합물과 플라보노이드 함량을 측정한 결과, 각각 115.42 ㎎/g, 23.07 ㎎/g이었다. EEEP의 DPPH radical 소거활성을 측정한 결과, IC50가 11.09 ㎍/ml로 뛰어난 항산화능을 보유한 것을 확인하였다. 또한 EEEP는 농도의존적으로 인체폐암세포주인 A549의 세포 성장을 저해하였으며, 세포 주기변화를 분석한 결과 A549 세포의 SubG1기 세포비율이 증가하는 것을 확인하였다. Annexin V 염색과 DAPI 염색으로 EEEP 처리에 의해 apoptotic 세포와 apoptotic body가 증가하는 것을 확인하였으며, 이러한 결과는 EEEP에 의해 A549 세포의 apoptosis가 유도되는 것을 시사한다. 또한 관련 단백질들의 발현변화를 분석한 결과, EEEP에 의해 Fas, p53, Bax의 발현이 증가하고 Bcl-2의 발현은 감소하였으며, caspase-8, -9와 caspase-3의 활성화를 통해 PARP가 분해되어 apoptosis가 유도되었음을 확인하였다. 이러한 결과들로부터 EEEP는 내인성 및 외인성 경로를 통한 apoptosis 유도에 의하여 A549 세포의 증식을 억제하는 항암활성을 보유하였음을 확인하였다. Euonymus porphyreus, a species of plant in the Celastraceae family, is widely distributed in East Asia, especially in Southern China. The botanical characteristics of E. porphyreus have been reported, but its antioxidative and anticancer activities remain unclear. In this study, we evaluated the antioxidative and anticancer effects of ethanol extracts of E. porphyreus (EEEP) and the molecular mechanism of its anticancer activity in human lung adenocarcinoma A549 cells. The total polyphenol and flavonoid compound contents from EEEP were 115.42 ㎎/g and 23.07 ㎎/g, respectively. EEEP showed significant antioxidative effects with a concentration at 50% of the inhibition (IC50) value of 11.09 ㎍/ml, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. EEEP showed cytotoxic activity by increasing the SubG1 cell population of A549 cells in a dose-dependent manner. Apoptosis in A549 cells treated with EEEP was evident due to increased apoptotic cells and apoptotic bodies, as detected by Annexin V and 4,6-diamidino-2-phenylindole (DAPI) staining, respectively. EEEP-induced apoptosis resulted in increased expression of the First apoptosis signal (Fas), p53, and Bax, with decreased expression of Bcl-2 and subsequent activation of caspase-8, -9, and caspase-3, leading to cleavage of poly (ADP-ribose) polymerase (PARP). Collectively, these results suggest that EEEP may exert an anticancer effect by inducing apoptosis in A549 cells through both intrinsic and extrinsic pathways.

인체 폐암 세포주 A549에서 Litsea populifolia 추출물의 항산화 및 항암활성 분석

진수정(Soojung Jin),오유나(You Na Oh),정현영(Hyun Young Jeong),윤희정(Hee Jung Yun),박정하(Jung-ha Park),권현주(Hyun Ju Kwon),김병우(Byung Woo Kim) 한국생명과학회 2019 생명과학회지 Vol.29 No.6

본 연구에서는 인체 폐암 세포인 A549를 사용하여 Litsea populifolia 에탄올 추출물(EELP)의 항산화 및 항암활성과 그 분자적 기전에 관하여 연구하였다. 먼저 EELP의 DPPH 라디칼 소거활성을 측정한 결과, IC50가 11.71 μg/ml로 유의적인 항산화활성을 보였다. 또한 EELP가 인체폐암세포주인 A549와 정상 폐세포인 IMR90의 세포증식에 미치는 영향을 알아본 결과, 정상세포의 생존율에는 거의 영향을 끼치지 않은 반면, EELP 농도의존적으로 A549 세포의 성장이 저해되었으며, 세포 주기 변화를 분석한 결과 EELP에 의해 A549 세포의 강력한 G1 arrest가 유도되는 것을 확인하였다. EELP에 의해 유도되는 G1 arrest는 세포주기 조절 인자인 Cyclin D1, Cyclin E, Cyclin-dependent kinase인 CDK2와 CDK6의 mRNA 발현 감소와 더불어 단백질 발현 감소와 연관되어 있었다. 또한 EELP 처리에 의한 CDK/Cyclin complex의 발현 저해는 DNA 손상에 의해 활성화되는 CHK2의 활성화 형태인 p-CHK2의 발현 증가에 따른 p53 인산화에 따른 활성화와 CDK 활성화 효소인 CDC25A 탈인산화효소의 인산화에 따른 저해에 의해 나타나는 결과로 사료된다. 이러한 결과들로부터 EELP는 두가지 경로인 p53-의존성과 p53-비의존성(ATM/CHK2/CDC25A/CDK2) 경로를 통해 A549의 G1 arrest를 유도하여 세포 증식을 억제하는 것으로 사료된다. 본 연구결과는 EELP가 폐암에 대한 새로운 항암활성 소재로서 사용될 수 있는 가능성을 시사하며, 또한 EELP의 세포주기 조절에 의한 항암기전을 이해하고 향후 지속적 연구를 하는 데 있어서 귀중한 기초자료로 사용될 수 있을 것이다. Litsea populifolia, a plant species of the Lauraceae family, is widely distributed in the tropical and subtropical areas of Asia. The phylogenetic relationships and botanical characteristics of L. populifolia have been reported; however, its anti-oxidative and anti-cancer activities remain unclear. In this study, we evaluated the anti-oxidative and anti-cancer effects of ethanol extracts of L. populifolia (EELP) together with the molecular mechanism of its anti-cancer activity in human lung adenocarcinoma A549 cells. EELP showed significant anti-oxidative effects with a 50% inhibitory concentration at 11.71 μg/ml, which was measured by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. EELP exhibited cytotoxic activity and induced cell cycle arrest at the G1 phase in A549 cells in a dose-dependent manner, whereas EELP did not have the cytotoxic effect on the normal human lung cell line IMR90. Treatment with EELP also resulted in a decreased expression of G1/S transition-related molecules—including cyclin-dependent kinase (CDK) 2, CDK6, cyclin D1, and cyclin E—both for the transcription and translation levels. EELP-induced G1 arrest was associated with the phosphorylation of checkpoint kinase 2 (CHK2), p53, cell division cycle 25 homolog A (CDC25A), and the reduction of CDC25A expression in A549 cells. Collectively, these results suggest that EELP may exert an anti-cancer effect by cell cycle arrest at the G1 phase through both p53-dependent and p53-independent (ATM/CHK2/CDC25A/CDK2) pathways in A549 cells.

A Novel All-trans Retinoid Acid Derivative N-(3-trifluoromethyl-phenyl)-Retinamide Inhibits Lung Adenocarcinoma A549 Cell Migration through Down-regulating Expression of Myosin Light Chain Kinase

Fan, Ting-Ting,Cheng, Ying,Wang, Yin-Feng,Gui, Shu-Yu,Chen, Fei-Hu,Zhou, Qing,Wang, Yuan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.18

Aim: To observe the effects of a novel all-trans retinoid acid (ATRA) derivative, N-(3-trifluoromethyl-phenyl)-retinamide (ATPR), on lung adenocarcinoma A549 cells and to explore the potential mechanism of ATPR inhibiting of A549 cell migration. Materials and Methods: The cytotoxicity of ATRA and ATPR on A549 cells was assessed using MTT assay. Wound healing assays were used to analyze the influences of ATRA, ATPR, ML-7 (a highly selective inhibitor of myosin light chain kinase (MLCK)), PMA (an activator of MAPKs) and PD98059 (a selective inhibitor of ERK1/2) on the migration of A549 cells. Expression of MLCK and phosphorylation of myosin light chain (MLC) were assessed by Western blotting. Results: ATRA and ATPR inhibited the proliferation of A549 cells in a dose- and time-dependent manner, and the effect of ATPR was much more remarkable compared with ATRA. Relative migration rate and migration distance of A549 cells both decreased significantly after treatment with ATPR or ML-7. The effect on cell migration of PD98059 combining ATPR treatment was more notable than that of ATPR alone. Moreover, compared with control groups, the expression levels of MLCK and phosphorylated MLC in A549 cells were both clearly reduced in ATRA and ATPR groups. Conclusions: ATPR could suppress the migration and invasion of A549 cells, and the mechanism might be concerned with down-regulating the expression of MLCK in the ERK-MAPK signaling pathway, pointing to therapeutic prospects in lung cancer.

Comparative Global Transcription Analysis of Hydrophilic Sapphyrin and Resveratrol on A549 Cell

장주선,장형진,정주영 한국바이오칩학회 2009 BioChip Journal Vol.3 No.4

Anti-cancer agents were studied to use a variety of natural product and chemical synthesis compounds, the effect elucidate cell cycle arrest, growth inhibitor, and death pathway factor expression and so on cancer cell. However molecular mechanisms, by which these anti-cancer agents kill and the extent to which cancer cell, including A549 cell, are resistant remains unclear. In previously DNA microarrays were utilized to analyze the genome-wide transcription changes in A549 cell after anti-cancer compounds (resveratrol, sapphyrin PCI-2050) exposure. In this study, we compared-cell eath pathway, cell cycle arrest, growth inhibition and so on-with both gene expression of sapphyrin (PCI-2050) and resveratrol that were treated on A549. And these investigated that resistance to any mechanism to A549 cell. Resveratrol treated A549 cell showed cell death pathway, cell cycle arrest, growth inhibition factor-related gene expression, and sapphyrin treated cell weren’t indicated gene expression of cell cycle arrest or growth inhibition. But it exhibited over-expression of c-jun on MAPK (JNK) pathway. Consequently, resveratrol resistance to cancer cell through several apoptotic pathways. But Sapphyrin may estimate to inhibit cancer cell by only certain MAPK (JNK) pathway. Anti-cancer agents were studied to use a variety of natural product and chemical synthesis compounds, the effect elucidate cell cycle arrest, growth inhibitor, and death pathway factor expression and so on cancer cell. However molecular mechanisms, by which these anti-cancer agents kill and the extent to which cancer cell, including A549 cell, are resistant remains unclear. In previously DNA microarrays were utilized to analyze the genome-wide transcription changes in A549 cell after anti-cancer compounds (resveratrol, sapphyrin PCI-2050) exposure. In this study, we compared-cell eath pathway, cell cycle arrest, growth inhibition and so on-with both gene expression of sapphyrin (PCI-2050) and resveratrol that were treated on A549. And these investigated that resistance to any mechanism to A549 cell. Resveratrol treated A549 cell showed cell death pathway, cell cycle arrest, growth inhibition factor-related gene expression, and sapphyrin treated cell weren’t indicated gene expression of cell cycle arrest or growth inhibition. But it exhibited over-expression of c-jun on MAPK (JNK) pathway. Consequently, resveratrol resistance to cancer cell through several apoptotic pathways. But Sapphyrin may estimate to inhibit cancer cell by only certain MAPK (JNK) pathway.

Effect of Withaferin A on A549 Cellular Proliferation and Apoptosis in Non-small Cell Lung Cancer

Cai, Yong,Sheng, Zhao-Ying,Chen, Yun,Bai, Chong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.4

Objective: To explore the effect of Withaferin A on A549 cellular proliferation and apoptosis in non-small cell lung cancer (NSCLC). Materials and Methods: NSCLC cell line A549 was selected to explore the effect of Withaferin A on A549 cellular proliferation, apoptosis and the PI3K/Akt signal pathway capable of regulating tumor biological behavior by assessment of cellular proliferation, cellular apoptotic rates and cellular cycling as well as by immuno-blotting. Results: Withaferin A could inhibit A549 cellular proliferation and the control rate was dosage-dependent (P<0.05), which also increased time-dependently with the same dosage of Withaferin A (P<0.05). The apoptotic indexes in A549 cells treated with 0, 2.5, 5.0, 10.0 and 20.0 ${\mu}mol{\cdot}L^{-1}$ Withaferin A for 48 h were significantly different (P<0.05). In addition, the apoptotic rates of each group in both early and advanced stages were higher than those in 0 ${\mu}mol{\cdot}L^{-1}$ (P<0.05), which were evidently higher after 48 h than those after 24 h (P<0.05). A549 cells treated by Withaferin A for 48 h were markedly lower in Bcl-2 level and obviously higher in Bax and cleaved caspase-3 levels than those treated by 0 ${\mu}mol{\cdot}L^{-1}$ Withaferin A (P<0.05), and there were significant differences among 5, 10 and 20 ${\mu}mol{\cdot}L^{-1}$ Withaferin A (P<0.05). The ratios of A549 cells treated by Withaferin A for 48 h in G0/G1 stage were higher than those in 0 ${\mu}mol{\cdot}L^{-1}$, while those in S and G2/M stages were obviously lower than those in G2/M stage, and there were significant differences in 5.0, 10.0 and 20.0 ${\mu}mol{\cdot}L^{-1}$ Withaferin A (P<0.05). Additionally, p-Akt/Akt values were in reverse association with dosage, and the differences were significant (P<0.05). Conclusion: Withaferin A can inhibit the proliferation and apoptosis of A549 cells by suppressing activation of the PI3K/Akt pathways.

사람 폐암 세포주에서 시스플라틴이나 독소루비신의 세포독성에 미치는 녹차 추출물의 영향

이병래(Byoung-Rai Lee),박재윤(Jae-Yoon Park),박평심(Pyoung-Sim Park) 한국식품영양과학회 2011 한국식품영양과학회지 Vol.40 No.5

항암 화학요법제의 항암작용을 증가시키거나, 부작용을 감소시켜 항암 치료를 효과적으로 할 수 있는 항암치료 보조제(modulator)에 대한 개발의 일환으로 녹차 추출물의 이용 가능성을 추정하기 위하여 사람 폐암 세포주인 A549 세포를 배양하여 시스플라틴과 독소루비신의 항암성에 미치는 녹차 추출물과 EGCG의 영향을 비교 관찰하였다. A549 세포에 독성을 나타나는 농도는 녹차 추출물 400 ㎍/mL, EGCG 300 ㎍/mL, 시스플라틴 10 ㎍/mL 및 독소루비신 8 ㎍/mL로, 녹차 추출물이 세포독성을 나타내는 농도는 시스플라틴이나 독소루비신에 비하면 낮았다. A549 세포에서 시스플라틴 10 ㎍/mL 이상의 농도에서 세포활성이 감소되었고, EGCG나 녹차 추출물 100 ㎍/mL를 첨가하면 시스플라틴 6 ㎍/mL 이상의 농도에서 세포활성이 감소되어 EGCG나 녹차 추출물 첨가로 시스플라틴의 세포독성이 증가되었다. A549 세포에서 독소루비신 8 ㎍/mL 이상의 농도에서 세포활성이 감소되었고, EGCG나 녹차 추출물 100 ㎍/mL를 첨가하면 독소루비신 4 ㎍/mL 이상의 농도에서 세포활성이 감소되어 EGCG나 녹차 추출물 첨가로 독소루비신의 세포독성이 증가되었다. A549 세포에서 녹차추출물 투여 후 p53 및 caspase-3에 대한 Western blot을 시행한 결과 p53및 caspase-3의 유전자 발현이 증가되었다. 이상의 실험결과 녹차추출물은 광범위 항암제 시스플라틴이나 독소루비신의 세포독성을 증강시키는 효과가 있고, 녹차추출물에 의한 p53이나 caspase-3 등과 같은 세포자살유도 단백질의 발현 증가는 녹차추출물에 의한 세포독성 증강효과와 연관이 있을 것으로 추측된다. 녹차추출물의 시스플라틴이나 독소루비신 세포독성 증강효과는 항암화학요법제의 용량을 늘리지 않고 항암력을 증대시킬 수 있기 때문에 항암화학요법 보조제로서 이용될 수 있는 가능성이 높은 것으로 생각되며, 이러한 효과를 규명하기 위한 연구가 필요할 것으로 사료된다. Tea extract (TE) has been shown to have anti-tumor properties in a wide variety of experimental systems. We evaluated green tea extract (GTE) as a biochemical modulator for the antitumor activity of cisplatin and doxorubicin in the treatment of human lung cancer A549 cells. Cells were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and two antibiotics (100 units/mL penicillin and 100 ㎍/mL streptomycin). Two types of TE, epigallocatechin galate (EGCG) and GTE, were used in this experiment. The cells were seeded at 1×10⁴ cells/well in the RPMI-1640 media with or without TE (100 ㎍/mL) and then treated with different concentrations of doxorubicin (0~14 ㎍/mL) or cisplatin (0~35 ㎍/mL). After incubation in 5% CO₂ at 37℃ for 24 hr, cell viability was determined with a MTT assay. We used a Western blot to detect the influence of EGCG and GTE on the expression of p53 and caspase-3 genes in the A549 cells. A549 cell viability decreased to 15% with a 10 ㎍/mL concentration of cisplatin, and to 21% with a 8 ㎍/mL concentration of doxorubicin, as measured with the MTT assay. However, pre-treatment of the cells with EGCG (100 ㎍/mL) or GTE (100 ㎍/mL) resulted in decreased cell viability with 6 ㎍/mL of cisplatin and 4 ㎍/mL of doxorubicin. There was no apparent change in cell viability between EGCG or GTE administration in cisplatin- or doxorubicin-induced cytotoxicity in A549 cells. The levels of p53 and caspase-3 in the A549 cells increased with both EGCG and GTE treatment. We found that GTE could potentially affect cisplatin- or doxorubicin-induced cytotoxicity of A549 cells, which may be useful in the chemotreatment of cancer.