• Asian Pacific Journal of Cancer Prevention
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• v.16 no.10
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• pp.4357-4361
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• 2015

Thunbergia Laurifolia Linn. (TL) is one of the most familiar plants in Thai traditional medicine that is used to treat various conditions, including cancer. However, the antitumor activity of TL or its constituents has never been reported at the molecular level to support the folklore claim. The present study was designed to investigate the antitumor effect of an aqueous extract of TL in human breast cancer cells and the possible mechanism(s) of action. An aqueous crude extract was prepared from dried leaves of TL. Folin-Ciocalteu colorimetric assays were used to determine the total phenolic content. Antiproliferative and cell cycle effects were evaluated in human breast adenocarcinoma MCF-7 cells by MTT reduction assay, cell growth inhibition, clonogenic cell survival, and flow cytometric analysis. Free radical generation by the extracts was detected using electron paramagnetic resonance spectroscopy. The exposure of human breast adenocarcinoma MCF-7 cells to a TL aqueous extract resulted in decreases in cell growth, clonogenic cell survival, and cell viability in a concentration-dependent manner with an $IC_{50}$ value of $843{\mu}g/ml$. Treatments with extract for 24h at $250{\mu}g/ml$ or higher induced cell cycle arrest as indicated by a significant increase of cell population in the G1 phase and a significant decrease in the S phase of the cell cycle. The capability of the aqueous extract to generate radical intermediates was observed at both high pH and near-neutral pH conditions. The findings suggest the antitumor bioactivities of TL against selected breast cancer cells may be due to induction of a G1 cell cycle arrest. Cytotoxicity and cell cycle perturbation that are associated with a high concentration of the extract could be in part explained by the total phenolic contents in the extract and the capacity to generate radical intermediates to modulate cellular proliferative signals.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.337-372
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• 1998

To evaluate the effects of Injinchunggantang-derivative on cell viability, cell cycle progression, and apoptosis, MTT assay, cell cycle analysis, Cpp32 protease assay, DNA fragnemtation assay, quantitative RT-PCR, and Western blotting were performed. The results were as followes. In MTT assay, etoposide+Injinchunggantang-derivative-treated cells as well as Injinchunggantang-derivative-treated cells showed higher viability than etoposide-treated cells with no time-concentration-dependence, which implied that Injinchunggantang-derivative has hepato-protective effect Cell cycle analysis showed that Injinchunggantang-derivative has no significant effect on the cell cycle. Cpp32 protease assav and DNA fragmentation assay Injinchunggantang-derivative carry inhibitory effects on apoptosis induction. It was suggested that Injinchunggantang-delivative might regulate the cell cycle, in particular $G_1$ checkpoint by blocking p53 and Watl pathway. Injinchunggantang-derivative inhibited the mRNA expressions of Cpp32, Fas, and Bcl-2, which could result in inhibition of apoptosis. These results imply that Injinchunggantang-derivative increases hepatocyte viability, and protects hepatocyte from damage by regulating the expression of genes associated with cell cycle and apoptosis, which explains the mechanism of the clinical effect of Injinchunggantang-derivative on liver diseases.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.1
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• pp.169-180
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• 2005

Objective : Spatholobus Suberectus Dunn stems, Chinese vine plants, have been used for the relief of menstrual disorders and rheumatic arthralgia. In this study, we investigated the antitumor effect of Spatholobus Suberectus Dunn on cervical cancer in vitro. Methods : HeLA cervical cancer cell lines were used as targets. We examined the effect of water extract from Spatholobus Suberectus Dunn on cell proliferation, cell cycle regulation and cell cycle-regulating gene expression. Further, we investigated the apoptotic effects of Spatholobus Suberectus Dunn on cervical cancer cell lines. Results : Spatholobus Suberectus Dunn significantly inhibited the proliferation of cervical cancer cell lines in a dose-dependent and time dependent manner. Fluorescence activated cell sorter (FACS) analysis indicated that Spatholobus Suberectus Dunn induced G1 cell cycle arrest. Spatholobus Suberectus Dunn enhanced the expression of $p21^{waf1}$ and $p27^{kip1}$ with cell cycle arrest. Further, Spatholobus Suberectus Dunn stimulated apoptosis via caspase3 pathway. Conclusions : These findings suggest that Spatholobus Suberectus Dunn is a candidate agent for the treatment of cervical cancer. p21waf1 and $p21^{waf1}$ and $p27^{kip1}$ may play an important role in Spatholobus Suberectus Dunn-induced cell cycle arrest and cell growth inhibition.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.4
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• pp.2343-2347
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• 2013

Objective: This investigation aimed to determine effects of celecoxib on the cell cycle kinetics of the gastric cancer cell line MGC803 and the mechanisms involved by assessing expression of cytochrome C and caspase-9 at the protein level. Methods: Cell proliferation of MGC803 was determined by MTT assay after treatment with celecoxib. Apoptosis was assessed using fluorescence staining and cell cycle kinetics by flow cytometry. Western blotting was used to detect the expression of caspase-9 protein and of cytochrome C protein in cell cytosol and mitochondria. Results: Celecoxib was able to restrain proliferation and induce apoptosis in a dose- and time-dependent manner, inducing G0/G1 cell cycle arrest, release of cytochrome C into the cytosol, and cleavage of pro-caspase-9 into its active form. Conclusion: Celecoxib can induce apoptosis in MGC803 cells through a mechanism involving cell cycle arrest, mitochondrial cytochrome C release and caspase activation.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8313-8319
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• 2016

We have recently shown that thymoquinone (TQ) has a potent cytotoxic effect and induces apoptosis via caspase-3 activation with down-regulation of XIAP in mouse neuroblastoma (Neuro-2a) cells. Interestingly, our results showed that TQ was significantly more cytotoxic towards Neuro-2a cells when compared with primary normal neuronal cells. In this study, the effects of TQ on cell-cycle regulation and the mechanisms that contribute to this effect were investigated using Neuro-2a cells. Cell-cycle analysis performed by flow cytometry revealed cell-cycle arrest at G2/M phase and a significant increase in the accumulation of TQ-treated cells at sub-G1 phase, indicating induction of apoptosis by the compound. Moreover, TQ increased the expression of p53, p21 mRNA and protein levels, whereas it decreased the protein expression of PCNA, cyclin B1 and Cdc2 in a dose-dependent manner. Our finding suggests that TQ could suppress cell growth and cell survival via arresting the cell-cycle in the G2/M phase and inducing apoptosis of neuroblastoma cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.4
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• pp.1-12
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• 2019

Objectives : The purpose of this study was to identify the anticancer effect of biological substances of ethylacetate(EtOAc) fraction from Orostachys japonicus(OJEF), their effect on human melanoma A375 cells and the related molecular mechanisms. Methods : The MTS assay was used to confirm the inhibition of cancer cell proliferation in A375 cells. And the $MUSE^{TM}$ analyzer was used to determine the ability of OJEF to induce cell cycle arrest. Western blotting was used to determine the changes in protein expression in A375 cells after treatment with OJEF. Results : OJEF showed cytotoxicity to A375 cells. And cell cycle arrest occurred in G1 phase and G2/M phase owing to inhibition of CDK1, cyclin B1, CDK4, and cyclin D, which are related to cell cycle regulation and cell division control. Conclusion : OJEF is effective in regulating cell cycle of human melanoma cells and thus can be a good theraputic agent to treat patients with melanoma.

• Journal of Life Science
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• v.16 no.5
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• pp.828-833
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• 2006

Maize (Zea mays L.) pistil cell death and stamen cell arrest are pivotal process on the sex determination, which diverges from bisexual state of floral meristem to unisexual state in staminate or pistillate floret. We investigated the temporal and spatial distribution of cell cycle gene expression during maize sex determination. The positive regulatory genes of cell cycle, cyclin A, cyclin B, cyclin dependent kinase (CDK) and Mad2 were highly expressed in the developing pistil and stamen but the expression was disappeared in the dying pistil and arresting stamens. In contrast, the negative regulatory genes of cell cycle, Wee1 and CDK inhibitor (CKI) were expressed in the arresting stamens in the wild-type ear and tasselseed2 mutant tassel, however, these genes were not detected in dying pistil although the cyclin B gene expression was disappeared. These results suggest that both the pistil cell death and stamen cell arrest process in maize sex determination are involved in cell cycle regulation, but the different expression patterns of negative regulatory cell cycle genes in the arresting stamens and aborting pistils suggest that the two processes may have distinctive modes of action.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.60-73
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• 2005

Objectives : The aim of this study was to evaluate the effects of Loranthus parasiticus Merr. on cell cycle and expression of related genes in HepG2 cells. Methods : The MTT assay, cell counting assay, $[^3H]-Thymidine$ incorporation assay, flow cytometric analysis, quantitative RT-PCR and western blot assay were studied. Results : In the water extract of Loranthus parasiticus Merr., inhibition of cell proliferation and DNA synthesis in HepG2 cells was seen. These inhibitory effects were due to inhibition of G l-S transition in cell cycle. After treatment with the extract, expression of cyclin D1(G1 check point related gene) was inhibited particularly in dose-dependent and time-dependent manners. Conclusion : These results suggest that the inhibition of cell cycle progression by Loranthus parasiticus Merr. in HepG2 cell is due to suppression of cyclin D1(G1 check point related gene) mRNA expression and protein synthesis.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.153-158
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• 2003

The genome-wide program of gene expression during the cell division cycle response to serum free media in chinese hamster ovary(CHO) cells was characterized using cDNA microarrays. Many transcripts of genes showed variation during the cell cycle. Characterization is critical step toward understanding the basic cell cycle processes and serum-free media role in CHO cells

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.495-498
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• 1994

Genistein, a inhibitor of the progression of G$_{1}$ and G$_{2}$ phase of the mammalian cell cycle, was discovered through a unique screening system, in which effects of microbial metabolites on the cycle progression of the cultured mouse mammalian carcinoma cell were monitored by flow cytometry. The inhibitor was extracted from the fermentation broth of Streptomyces sp. ZF10 with ethyl acetate, and purified by silica gel column chromatography and HPLC.