• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.769-773
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• 2014

Fangchinoline (Fan) inhibits cell proliferation and induces apoptosis in several cancer cell lines. The effects of Fan on cell growth and proliferation in breast cancer cells remain to be elucidated. Here, we show that Fan inhibited cell proliferation in the MDA-MB-231 breast cancer cell line through suppression of the AKT/Gsk-3beta/cyclin D1 signaling pathway. Furthermore, Fan induced apoptosis by increasing the expression of Bax (relative to Bcl-2), active caspase 3 and cytochrome-c. Fan significantly inhibited cell proliferation of MDA-MB-231 cells in a concentration and time dependent manner as determined by MTT assay. Flow cytometry analysis demonstrated that Fan treatment of MDA-MB-231 cells resulted in cell cycle arrest at the G1 phase, which correlated with apparent downregulation of both mRNA and protein levels of both PCNA and cyclin D1. Further analysis demonstrated that Fan decreased the phosphorylation of AKT and GSK-3beta. In addition, Fan up-regulated active caspase3, cytochrome-c protein levels and the ratio of Bax/Bcl-2, accompanied by apoptosis. Taken together, these results suggest that Fan is a potential natural product for the treatment of breast cancer.

• 대한약침학회지
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• 제7권2호
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• pp.5-17
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• 2004

Objective : In order to measure the efficacy of cultivated wild ginseng distilled herbal acupuncture by concentration level, we've treated A549 human lung cancer lines with different concentrations of cultivated wild ginseng distilled herbal acupuncture and examined mRNA and proteins which take parts in apoptosis. Methods : A549 human lung cancer lines were treated with various concentration levels of cultivated wild ginseng distilled herbal acupuncture and cell toxicity was carefully examined. From the analysis of DNA fragmentation, RT-PCR and Western blot, manifestation of mRNA and proteins which are associated with apoptosis were inspected. Results : The following results were obtained on apoptosis of A549 human lung cancer lines after administering various concentration levels of cultivated wild ginseng distilled herbal acupuncture. 1. Measuring cell toxicity of lung cancer cells, strong cell toxicity was detected at high concentration level(1000ul, 1200ul), but no consistent concentration dependent reliance was detected. 2. Through DNA fragmentation, we were able to confirm cell destruction in all groups. 3. Experiment groups treated with cultivated wild ginseng distilled herbal acupuncture showed inhibition of Bcl-2 and COX-2 at mRNA and Protein level, whileas increase of Bax was shown. 4. Manifestation of p21, p53, Cyclin E, and Cyclin Dl were confirmed in all groups. 5. Extrication of Cytochrome C was detected at all groups, as well as increased activity of the enzyme caspase-3 and caspase-9, and PARP fragmentation were confirmed. Conclusion : According to the results, we can carefully deduce cell destruction of A549 human lung cancer lines were induced by Apoptosis. At the fixed level, cultivated wild ginseng distilled herbal acupuncture showed decrease of Bcl-2 and COX-2, as well as increase of Bax. Since cultivated wild ginseng distilled herbal acupuncture increases manifestation of p21, p53, Cyclin E, and Cyclin Dl, it affects cellular cycle and through these phenomena, we can consider extrication of Cytochrome C, increase of caspase, and PARP fragmentation are the results.

• Bulletin of the Korean Chemical Society
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• 제28권2호
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• pp.211-219
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• 2007

The subtle but significant differences and thereby the lack of consensus in active site structures among the crystal structures of cyclin-dependent kinase 2 (CDK2) has hampered structure-based drug design. In this study, we devised a simple but effective ‘mutation, pharmacophore-guided docking, followed by mutation' strategy to generate an “average” CDK2 structure, which was used for ligand docking study to successfully reproduce 30 out of 32 X-ray ligand positions within 2.0 A of heavy atom RMSD. This novel docking method was applied for structure-based 3D QSAR with CoMSIA study of a series of structurally related ligands, which showed a good discrimination between CDK2 binders and nonbinders.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7181-7185
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• 2014

Cyclin D1 is an important positive regulator of the G1/S phase of the cell cycle. We investigated the association between the CCND1 G870A polymorphism and susceptibility to papillary thyroid cancer in Turkish people. This study covered 102 patients with papillary thyroid cancer and 174 healthy controls. CCND1 genotyping was determined by the PCR-RFLP method. We found that the A allele frequency was higher in the cases than in the controls (p=0.042). On stratification analysis, papillary thyroid cancer risk was significantly elevated in individuals older than 45 years with the A allele (OR=1.91, 95% CI, 1.09-3.35, p=0.024) and in females with the A allele (OR=1.73, 95% CI, 1.06-2.84, p=0.029), compared to the G allele. According to the subject age, there was an increased papillary thyroid cancer risk for the individuals older than 45 years with the AA genotype (OR=2.28, 95% CI, 1.02-5.13, p=0.046) compared to the AG+GG combined genotypes. In conclusion, it is suggested that the CCND1 G870A polymorphism may contribute to the susceptibility to papillary thyroid cancer, especially in those who were older subjects ($45{\leq}$ years old) and female, in the Turkish population.

• 생명과학회지
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• 제23권9호
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• pp.1109-1117
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• 2013

본 연구에서는 인체 폐암 세포인 A549를 사용하여 택란 메탄올 추출물의 항암활성과 그 분자적 기전에 관하여 연구하였다. 먼저 택란 추출물이 A549의 세포증식에 미치는 영향을 알아본 결과 처리 농도 및 시간 의존적으로 A549의 성장이 저해되었으며, 세포 주기 변화를 분석한 결과 강력한 G1 arrest가 유도되는 것을 확인하였다. 이러한 택란 추출물에 의한 G1 arrest는 세포주기 조절 단백질인 Cyclin D1, Cyclin E 및 Cyclin-dependent kinase인 CDK2, CDK4, CDK6의 발현 감소와 연관되어 있었다. 또한 택란 추출물에 의한 CDK/Cyclin complex의 발현 저해는 DNA 손상에 의해 활성화되는 CHK2의 활성화 형태인 p-CHK2의 발현 증가에 따른 CDK 활성화 효소인 Cdc25A phosphatase의 발현 억제에 의해 나타나는 결과로 사료된다. 반면 종양억제유전자인 p53 및 CDK 억제제인 p21과 p27의 발현량은 증가되지 않았다. 이러한 결과들로부터 택란 추출물은 DNA damage에 의한 ATM/CHK2/Cdc25A/CDK2 pathway를 통해 A549의 G1 arrest를 유도하여 세포의 증식을 억제할 것으로 판단되며, 이때 택란 추출물에 의해 유도되는 G1 arrest는 p53 비의존적인 경로일 것으로 사료된다. 본 연구결과는 택란이 Cdc25A를 target으로 하는 새로운 항암활성 소재로서 사용될 수 있는 가능성을 시사한다. 또한 본 연구결과는 택란 추출물의 세포주기 조절에 의한 항암기전을 이해하고 향후 지속적 연구를 하는 데 있어서 귀중한 기초자료로 사용될 수 있을 것이다.

• Natural Product Sciences
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• 제19권2호
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• pp.155-159
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• 2013

Honokiol, a naturally occurring neolignan mainly found in Magnolia species, has exhibited a potential anti-proliferative activity in human cancer cells. However, the growth inhibitory activity against hepatocellular carcinoma cells and the underlying molecular mechanisms has been poorly determined. The present study was designed to examine the anti-proliferative effect of honokiol in SK-HEP-1 human hepatocellular cancer cells. Honokiol exerted anti-proliferative activity with cell-cycle arrest at the G0/G1 phase and sequential induction of apoptotic cell death. The cell-cycle arrest was well correlated with the down-regulation of checkpoint proteins including cyclin D1, cyclin A, cyclin E, CDK4, PCNA, retinoblastoma protein (Rb), and c-Myc. The increase of sub-G1 peak by the higher concentration of honokiol ($75{\mu}M$) was closely related to the induction of apoptosis, which was evidenced by decreased expression of Bcl-2, Bid, and caspase-9. Hohokiol was also found to attenuate the activation of signaling proteins in the Akt/mTOR and ERK pathways. These findings suggest that the anti-proliferative effect of honokiol was associated in part with the induction of cell-cycle arrest, apoptosis, and dow-nregulation of Akt/mTOR signaling pathways in human hepatocellular cancer cells.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.811-825
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• 2002

Recently, many natural medicines, whose advantages are less side effects and possibility of long-term use, have been studied for their capacity, their anti-bacterial and anti-inflammatory effects and regenerative potential of periodontal tissues. Cervi Parvum Comu(CPC) have been traditionally study as an hale, growth. hematogenous, anti-aging, hack pain in Eastern medicine. The purpose of present study was to investigate the effects of CPC extract on cell cycle progression and its molecular mechanism in human fetal osteoblasts. CPC extracts (10 ${\mu}g/ml$) increased cell proliferation in the human fetal osteoblasts compared to non-supplemented control. There was no significant change in the G1 and S phase, hut a increase in the G2/M phase in 10 ${\mu}g/ml$ and 100 ${\mu}g/ml$ of CPC extracts group as compared to non-supplemented control. The protein expression of cyclin E, cdk 2, cycln D, cdk 4, and cdk 6 was higher than that of control group. The level of p21 was lower than that of control. But that of pRb and pl6 was not distinguished from control. These results indicate that the increase of cell proliferation by CPC extracts may be due t o the increased expression of cyclin E, cdk 2, cyclin D, cdk 4 and cdk 6, and the decreased expression of p21 in human fetal osteoblasts.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1055-1060
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• 2003

Lysophosphatidic acid (LPA) is a well-known mitogen in various cell types. However, we found that LPA inhibits melanocyte proliferation. Thus, we further investigated the possible signaling pathways involved in melanocyte growth inhibition. We first examined the regulation of the three major subfamilies of mitogen-activated protein (MAP) kinases and of the Akt pathway by LPA. The activations of extracellular signal-regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK) were observed in concert with the inhibition of melanocyte proliferation by LPA, whereas p38 MAP kinase and Akt were not influenced by LPA. However, the specific inhibition of the ERK or JNK pathways by PD98059 or D-JNKI1, respectively, did not restore the antiproliferative effect. We next examined changes in the expression of cell cycle related proteins. LPA decreased cyclin $D_1 and cyclin D_2$ levels but increased $p21^{WAF1/CIP1}$ (p21) and $p27^{KIP1}$ (p27) levels, which are known inhibitors of cyclin-dependent kinase. Flow cytometric analysis showed the inhibition of DNA synthesis by a reduction in the S phase and an increase in the $G_0/G_1$ phase of the cell cycle. Our results suggest that LPA induces cell cycle arrest by regulating the expressions of cell cycle related proteins.

• Journal of the Korean Wood Science and Technology
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• 제46권2호
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• pp.166-177
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• 2018

Plant essential oils are defined as fragrant volatile oils extracted from leaves, stems, fruits, flowers, and roots of a plant. Such oils are composed of multiple components and multiple functions. By accumulation of inductive information, various plant essential oils have been studied for using in therapeutic medicine for various diseases. Despite of the apparent advantages of essential oils as a source of therapeutic medicines, plant essential oils have many limitations, including cytotoxic side effects. Therefore, it is necessary to evaluate the toxicity and the mechanisms of cytotoxicity of such oils. In this study, we evaluated the cytotoxicity to human-derived cell lines of 10 plant essential oils provided by National Institute of Forest Science (i.e., Larix kaempferi; Abies holophylla; Zanthoxylum ailanthoides; Pinus parviflora; Tsuga sieboldti; Chamaecyparis pisifera; Cryptomeria japonica; Pinus densiflora; Illicium anisatum; Pinus thunbergii). Cytotoxicity evaluations were accomplished by using CCK-assays and PCR-based cytotoxicity-related marker gene analyses with A549 cell line, and the Detroit551 cell line which are lung and skin cell line. The genes were analyzed included caspase-3 has a role in cell apoptosis, and the other cyclinA, cyclinB, cyclinD, and cyclinE regulated cell cycling for the cell proliferation. By examining the five cytotoxicity-related marker genes by performing real-time PCR and examined the cytostatic gene regulation associated with the various essential oils. The results of this study showed that the degree of cytotoxicity and the cytostatic gene regulation which could give precious information for using the plant essential oil for the clinical usages.

• 대한두경부종양학회지
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• 제27권1호
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• pp.59-65
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• 2011

Background and Objectives : Papillary thyroid carcinoma(PTC) frequently metastasize to the regional neck, however, lateral neck lymph node metastasis is less common. The aim of this study is to investigate clinical and immunohistochemical features of PTC with lateral LN metastasis, and determine the predictive factors for lateral LN metastases. Material and Methods : We undertook a retrospective study of 83 patients treated between January 2007 and December 2009 for PTC by thyroidectomy with or without lateral neck dissection. The following criteria were used to study the clinical predictive value of lateral LN. metastases : sex, age, tumor size, multifocality, extracapsular spread(ECS) and lymphovascular emboli. Immunohistochemical staining for VEGF-A, VEGF-C, Bax, Bcl-2, Cyclin D1, Cyclin E, $p27^{kip1}$ and $p57^{kip2}$ was performed, and quantified blindly by three pathologists who had no clinical information of the patients. Immunohistochemical expression was scored as high(>50% of cells stained) or low(0-49%). Results : With use of univariate and multivariate analysis, tumor size(>2cm) and ECS were independent correlates of lateral LN metastasis in PTC. Expression of VEGF-C, Bax, and Cyclin D1 in the PTC with lateral LN metastasis was scored higher than in PTC without lateral LN metastasis(p<0.05). Conclusion : The important risk factors for lateral LN metastasis in PTC are primary tumor size and the presence of ECS. And expression of VEGF-C, Bax and cyclin D1 may be considered of lateral LN metastatic potential in PTC.