• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.154.2-154.2
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• 2003

Mammalian cell cycles are tightly regulated by cyclins, cyclin dependent kinase (CDK), Retinoblatoma (Rb) protein, and cellular CDK inhibitors (CDKI). Cyelin dependent kinases (CDK) are key enzymes regulating eukaryotic cell cycle. And also it is recognized that the abnormal increase of CDK activities is one of the common events in human cancer and CDK inhibitors have therapeutic values in cancer treatment. Until now it is known that over 10 different CKDs participate in cell cycle regulation. (omitted)

• 대한임상검사과학회지
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• 제56권1호
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• pp.73-84
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• 2024

펄스 전자기장(pulsed electromagnetic field, PEMF)은 여러 항암제의 항암 효과를 향상시키는 것으로 알려져 있고 독소루비신(doxorubicin, DOX)은 유방암을 포함한 다양한 종류의 악성 종양을 치료하는 데 사용되는 항암제이다. 본 연구는 PEMF가 MCF-7 유방암 세포에 대한 DOX의 항암 효과 증진 여부를 조사하고 관련기전을 규명하기 위해 진행되었다. 본 연구팀은 DOX와 PEMF를 동시에 처리하면 DOX 단독 처리에 비해 MCF-7 유방암 세포의 생존율 감소가 더 커지는 것을 확인하였다. PEMF는 cyclin-dependent kinase 2의 인산화와 p53, p21, 사이클린 E2 및 polo like kinase 1의 단백질 발현에 영향을 주어 DOX 처리에 의한 G1 세포주기 정지를 더욱 증가시켰다. 또한, PEMF는 DOX 처리에 의한 Fas와 Bcl-2-associated X의 증가, myeloid leukemia 1과 survivin의 감소, 카스파제(caspase)-8/9/7의 활성 및 poly (adenosine diphosphate-ribose) polymerase 절단을 더욱 증가시켰다. 이러한 연구결과를 바탕으로, 본 연구팀은 PEMF는 DOX 처리에 의한 G1 세포주기 정지와 카스파제 의존적 세포자멸사를 더욱 증가시켜 DOX 처리에 의한 MCF-7 세포의 생존율 감소를 더욱 증진시킴을 확인할 수 있었다.

• 약학회지
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• 제46권1호
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• pp.18-23
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• 2002

Ailanthus altissima has been used to settle an upset stomach, to alleviate a fever and as an insecticide. We reported that the water extract of A. altissima induced apoptotic cell death in Jurkat T-acute Iymphoblastic leukemia cells. Here, we showed the dose-dependent inhibitions of cell viability by the extract, as measured by cell morphology. The cell cycle control genes are considered to play important roles in tumorigenesis. The purpose of the present study is also to investigate the effect of A. altissima on cell cycle progression and its molecular mechanism in the cells. The level of p21 protein was increased after treatment of the extract, whereas both Bcl-2 and Bax protein levels were not changed. These results suggest that A. altissima induces apoptotic cell death via p21-dependent signaling pathway in Jurkat cells which delete wild type p53. Gl checkpoint related gene products tested (cyclin D3, cyclin dependent kinase 4, retinoblastoma, E2Fl) were decreased in their protein levels in a dose-dependent manner after treatment of the extract Taken together, these results indicate that the increase of apoptotic cell death by A. altissima may be due to the inhibition of cell cycle in Jurkat cells.

• Biomolecules & Therapeutics
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• 제26권3호
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• pp.328-334
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• 2018

Because of the unsatisfactory treatment options for breast cancer (BC), there is a need to develop novel therapeutic approaches for this malignancy. One such strategy is chemotherapy using non-toxic dietary substances and botanical products. Studies have shown that Panduratin A (PA) possesses many health benefits, including anti-inflammatory, anti-bacterial, anti-oxidant and anticancer activities. In the present study, we provide evidence that PA treatment of MCF-7 BC cells resulted in a time- and dose-dependent inhibition of cell growth with an $IC_{50}$ of $15{\mu}M$ and no to little effect on normal human MCF-10A breast cells. To define the mechanism of these anti-proliferative effects of PA, we determined its effect critical molecular events known to regulate the cell cycle and apoptotic machinery. Immunofluorescence and flow cytometric analysis of Annexin V-FITC staining provided evidence for the induction of apoptosis. PA treatment of BC cells resulted in increased activity/expression of mitochondrial cytochrome C, caspases 7, 8 and 9 with a significant increase in the Bax:Bcl-2 ratio, suggesting the involvement of a mitochondrial-dependent apoptotic pathway. Furthermore, cell cycle analysis using flow cytometry showed that PA treatment of cells resulted in G0/G1 arrest in a dose-dependent manner. Immunoblot analysis data revealed that, in MCF-7 cell lines, PA treatment resulted in the dose-dependent (i) induction of $p21^{WAF1/Cip1}$ and p27Kip1, (ii) downregulation of Cyclin dependent kinase (CDK) 4 and (iii) decrease in cyclin D1. These findings suggest that PA may be an effective therapeutic agent against BC.

• Journal of Acupuncture Research
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• 제20권3호
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• pp.45-62
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• 2003

Objective : To examine the effects of Beevenom on the cell proliferation of human breast carcinoma cell line MCF-7, we performed various experiments such as does-dependent effect of Beevenom on cell proliferation and viability, morphological changes, and alterations of apoptosis/cell cycle-regulatory gene products. Methods : Beevenom induced cell viability and proliferation of MCF-7 cells in a concentration-dependent manner. The anti-proliferative effect by Beevenom treatment in MCF-7 cells was associated with morphological changes such as membrance shrinking and cell rounding up. Results : Beevenom induced apoptotic cell death in a concentration-dependent manager, which was associated with degradation of ${\beta}$-catenin, an apoptotic target protein. Beevenom induced the Bax expressions, a pro-apoptotic gene, both in protein and mRNA levels, however, the levels of Bcl-$X_{S/L}$ expression, an anti-apoptotic gene, were down-regulated in Beevenom-treated cells. Western blot analysis and RT-PCT data revealed that the levels of cyclin of B1 protein and cyclin E mRNA were reduced by Beevenom treatment in MCF-7 cells, respectively, where as the expression of tumor suppressor p53 and cyclin dependent kinase inhibitor p21 mRNA were markedly increased in a concentration-dependent fashion. Conclusions : Taken together, these findings suggest that Beevenom induced inhibition of human breast cancer cell proliferation is associated with the induction of apoptotic cell death and Beevenom may have therapeutic potential in human breast cancer.

• 생명과학회지
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• 제16권7호
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• pp.1229-1234
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• 2006

배지에 tetracycline이 없으면 사멸하는 평활근세포 (FADD-SMC)를 이용하여 metalloproteinase (MMP)와 사멸세포의 상관관계를 조사하였다. FADD-SMC를 tetracycline이 없는 배지에서 배양하는 경우 핵이 조각으로 잘라졌고 인산화한 p38과 ERK가 증가하고 MMP-9의 발현과 활성이 증가한 반면, cyclin D와 cyclin D의 발현은 감소하였다. 그리고 죽는 FADD-SMC에서 MMP-9의 발현은 immunofluorescen로 재확인하였다. MMP-9의 증가는 MAPK 억제제인 PD098059와 p38 MAPK 억제제인 SB203S80에 의하여 감소하였다. 그리고 MMP 억제제인 BB94는 FADD-SMC의 사멸을 감소시켰다.

• 생명과학회지
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• 제29권6호
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• pp.679-687
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• 2019

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, $IC_{50}$가 $11.71{\mu}g/ml$로 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.

• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.1045-1050
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• 2002

The aims of this study were to detect spontaneously occurring apoptosis in cultured porcine ovarian cells, to examine the role of growth hormone (GH), tyrosine kinase (TK), protein kinase G (PKG) and cyclin-dependent kinase (CDK) in the control of this process, and to determine whether the effect of GH on apoptosis is mediated by TK-, PKG- and cdc2-dependent intracellular mechanisms. We studied the action of pGH (10 ng/ml), blockers of TK (genistein, lavendustin, both 100 ng/ml), PKG (Rp-Br-PET-cGMPS, 50 nM; KT5823, 100 ng/ml) and CDK (olomoucine, $1{\mu}g/ml$), as well as combinations of GH with these blockers, on the onset of apoptosis in cultured granulosa cells isolated from antral (3-6 mm) porcine follicles. The functional characteristics of an early apoptotic event, DNA fragmentation, were determined using terminal deoxynucleotidyltransferase (TdT)-mediated dUTP nick end labelling (TUNEL), whilst morphological signs of advanced apoptosis such as pyknosis, chromatin marginalization, shrinkage and fragmentation of nucleus, were detected using routine light microscopy. After culture, some ovarian granulosa cells exhibited DNA fragmentation, which in some cases was associated with morphological apoptosis-related changes (pyknosis, shrinkage and fragmentation of the nucleus). GH significantly reduced the proportion of TUNEL-positive cells. Neither TK nor CDK blockers when given alone, significantly affected the percentage of TUNEL-positive cells although both PKG blockers significantly increased this index. Furthermore, TK and PKG blockers given together with GH, prevented or reversed the inhibitory effect of GH on apoptosis, whilst the CDK blocker olomoucine promoted it. These observations demonstrate apoptosis in porcine ovaries and suggest the involvement of GH, TK, PKG and CDK in the control of this process. They also suggest that the effect of GH on ovarian apoptosis is mediated or regulated by multiple signalling pathways including TK-, PKG- and CDK-dependent intracellular mechanisms.

• The Korean Journal of Physiology and Pharmacology
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• 제22권2호
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• pp.193-201
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• 2018

Connective tissue growth factor (CTGF) is a novel fibrotic mediator, which is considered to mediate fibrosis through extracellular matrix (ECM) synthesis in diabetic cardiovascular complications. Statins have significant immunomodulatory effects and reduce vascular injury. We therefore examined whether fluvastatin has anti-fibrotic effects in vascular smooth muscle cells (VSMCs) and elucidated its putative transduction signals. We show that advanced glycation end products (AGEs) stimulated CTGF mRNA and protein expression in a time-dependent manner. AGE-induced CTGF expression was mediated via ERK1/2, JNK, and Egr-1 pathways, but not p38; consequently, cell proliferation and migration and ECM accumulation were regulated by CTGF signaling pathway. AGE-stimulated VSMC proliferation, migration, and ECM accumulation were blocked by fluvastatin. However, the inhibitory effect of fluvastatin was restored by administration of CTGF recombinant protein. AGE-induced VSMC proliferation was dependent on cell cycle arrest, thereby increasing G1/G0 phase. Fluvastatin repressed cell cycle regulatory genes cyclin D1 and Cdk4 and augmented cyclin-dependent kinase inhibitors p27 and p21 in AGE-induced VSMCs. Taken together, fluvastatin suppressed AGE-induced VSMC proliferation, migration, and ECM accumulation by targeting CTGF signaling mechanism. These findings might be evidence for CTGF as a potential therapeutic target in diabetic vasculature complication.

• 한국식품영양과학회지
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• 제31권2호
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• pp.329-332
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• 2002

본 연구는 사람의 간암세포인 HepG2 세포를 대상으로 강력한 암 예방 효과물질을 함유하고 있을 것으로 추측되는 향버섯 메탄올 추출물의 암세포 성장 저해 효과를 검토하고 또한 암세포 성장 억제 효과의 분자생물학적 기전을 파악하기 위하여 암세포주의 세포주기 조절인자들의 발현을 조사하였다. 향버섯 메탄을 추출물의 HePG2세포에 대한 성장 저해 효과를 MTT assay로 검토한 결과 높은 암세포 성장 저해 효과를 나타내었으며 사람의 정상 간세포인 Chang cell에서는 세포독성이 나타나지 않았다. 또한 향버섯 추출물의 작용으로 HepG2 세포에서 cyclin A와 Dl 단백질의 발현이 억제 되었으며 cyclin Bl 단백질의 발현은 증가하는 경향을 나타내었다. 그리고 암 억제 단백질인 p53의 발현은 전반적으로 증가되었으며, 이와 대조적으로 PCNA 단백질은 감소하는 경향을 나타내었고 세포분열 억제 단백질 p27의 발현은 증가 하는 경향을 나타내었다. 이러한 결과로 볼 때 향버섯 메탄올 추출물은 간암세포의 세포주기 중 Gl기 에서 S기로의 진행을 조절하는 인자인 cyclin A와 cyclin Dl 발현을 억제시키고 p53, p27 단백질을 활성화 시킴과 동시에 PCNA 작용을 억제 함으로써 세포주기 중 Gl/S기 차단을 유도하여 암세포 증식을 억제한 것으로 추정된다.