• 한국환경성돌연변이발암원학회지
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• 제22권1호
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• pp.54-59
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• 2002

We have used cDNA microarray hybridization to identify gene regulated in response to gamma-irradiation in U-937 cell. The cDNA-chip was composed entirely of 1,000 human cancer related gene including apoptosis and angiogenesis etc. In gamma-irradiated U-937 cell, highly charged protein, ribosomal protein L32, four and a half LIM domains 3, lipocalin 2 (oncogene 24p3) and interleukin 15, ataxia telangiectasia mutated (includes complementation groups A, C and D) genes showed increased level of its transcription, and cell division cycle 25A, dihydrofolate reductase, topoisomerase (DNA) II beta(180kD), kinase suppressor of ras and strarigin genes showed reduced level of its transcription compared to untreated U-937 cell. The significant change of level of transcription was not found in well-known ionizing radiation(IR)-responsive gene, such as transcription factor TP53 and p53 related gene, except ataxia telangiectasia mutated gene.

• 생약학회지
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• 제33권1호통권128호
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• pp.29-34
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• 2002

Albizzia julibrissin belonging to the family Leguminosae has been used for the treatment of contusion, sore throat, amnesia, and insomnia in Oriental traditional medicine. The water extract of A. julibrissin induced apoptosis in Jurkat T-acute lymphoblastic leukemia (ALL) cells as measured by cell morphology. The capability of this herb medicine to induce apoptosis was associated with proteolytic cleavage of specific target protein such as beta-catenin protein suggesting the possible involvement of caspases. The purpose of the present study is also to investigate the effect of A. julibrissin on cell cycle progression. Our results showed that GI checkpoint related gene products (cyclin D1, cyclin dependent kinase 4, retinoblastoma, E2F1) were decreased in their protein levels in a dose-dependent manners after treatment of the extract. These results indicate that the increase of apoptotic cell death by A. julibrissin may be due to the inhibition of cell cycle progression in wild type p53-lacking Jurkat cells.

• International Journal of Oral Biology
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• 제45권4호
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• pp.152-161
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• 2020

D-pinitol is an analog of 3-methoxy-D-chiro-inositol found in beans and plants. D-pinitol has anti-inflammatory, antidiabetic, and anticancer effects. Additionally, D-pinitol induces apoptosis and inhibits metastasis in breast and prostate cancers. However, to date, no study has investigated the anticancer effects of D-pinitol in oral cancer. Therefore, in this study, whether the anticancer effects of D-pinitol induce apoptosis, inhibit the epithelial-to-mesenchymal transition (EMT), and arrest cell cycle was investigated in squamous epithelial cells. D-pinitol decreased the survival and cell proliferation rates of CAL-27 and Ca9-22 oral squamous carcinoma cells in a concentration- and time-dependent manner. Evidence of apoptosis, including nuclear condensation, poly (ADP-ribose) polymerase, and caspase-3 fragmentation, was also observed. D-pinitol inhibited the migration and invasion of both cell lines. In terms of EMT-related proteins, E-cadherin was increased, whereas N-cadherin, Snail, and Slug were decreased. D-pinitol also decreased the expression of cyclin D1, a protein involved in the cell cycle, but increased the expression of p21, a cyclin-dependent kinase inhibitor. Hence, D-pinitol induces apoptosis and cell cycle arrest in CAL-27 and Ca9-22 cells, demonstrating an anticancer effect by decreasing the EMT.

• BMB Reports
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• 제32권4호
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• pp.379-384
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• 1999

Taxol, a natural product with significant anti-tumor activity, stabilizes microtubules and arrests cells in the G2/M phase of the cell cycle. It has been reported that taxol has additional effects on the cell such as an increase in tyrosine phosphorylation of proteins and activation of mitogen-activated protein (MAP) kinase. This phosphorylated kinase translocates into the nucleus and phosphorylates its substrate c-jun, c-fos, ATF2, and ATF3. The MAP kinase family is comprised of key regulatory proteins that control the cellular response to both proliferation and stress signals. First examination was cytotoxicity and apoptosis-induced concentration with paclitaxel in HeLa cell. A half-maximal inhibition of cell proliferation ($IC_{50}$) occurred at 13 nM paclitaxel. When DNA fragmentation was analyzed by agarose gel electrophoresis, a nucleosomal ladder became evident 24 h after a taxol (50 nM) addition to the cells. In addition, an apoptotic body was detected by electron microscopy. Taxol-treated cells were arrested at the S phase at 10 nM. Treatment of 50 nM taxol activated the extracellular signal-regulated protein kinase (ERK1), and a fraction of the activated MAP kinases entered the nucleus. It was also discovered that nucleus substrates c-jun was phosphorylated and activated in the cell. The activated ERK1 could subsequently translocate into the nucleus and phosphorylate its substrate c-jun as well. This study suggests that taxol-induced apoptosis might be related with signal transduction via MAP kinases.

• Journal of Applied Biological Chemistry
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• 제57권2호
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• pp.113-122
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• 2014

A5E is complex of several medicinal herb ethanol extracts. The aim of this study is investigating the anticancer effect for non-small cell lung cancer. The antitumor effects of A5E on NCI-H460 were examined by regulation of cell proliferation, apoptosis, cell cycle arrest, mitochondrial membrane potential (${\Delta}{\Psi}_m$), and apoptosis-related protein. Cell proliferation was measured by MTS assay. Apoptosis induced by A5E was confirmed by Annexin V-fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) staining, and cell cycle arrest was measured by PI staining. NF-${\kappa}B$ translocation was detected by immunofluorescence and MMP (${\Delta}{\Psi}_m$) was measured by JC-1 staining. The expression of extrinsic pathway molecules such as FasL and FADD were elevated, and procaspase-8 was processed by A5E. In addition, intrinsic pathway related molecules were altered. The Bcl-2 and Bcl-xl levels decreased, Bax increased, and cytochrome C was released. In addition, the mitochondrial membrane potential collapsed, and caspase-3 and poly-(ADP-ribose) polymerase were processed by A5E. Moreover, A5E affected the cellular survival pathway involving phosphatidylinositol 3-kinase (PI3K)/Akt and NF-${\kappa}B$. PI3K and Akt were downregulated, also NF-${\kappa}B$ expression was decreased, and nuclear translocalization was inhibited by A5E. These results suggested that A5E delays proliferation, inhibit cell cycle progression and induce apoptosis in human lung cancer cell. We conclude that A5E is a potential anticancer agent for human lung carcinoma.

• 대한본초학회지
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• 제36권4호
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• pp.1-7
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• 2021

Objective : As more and more people are interested in appearance in modern society, the increasing number of hair loss population can have an important impact on psychological and social problems such as depression and inappropriate interpersonal symptoms. Therefore, much research is being done on treatments for alopecia using herbal extracts with relatively few side effects. This study was investigated about the effect of Achyranthis Radix (AR) extract with ethanol solvent on hair growth. Methods : We determined the promoting efficacy of AR-ethanol extract compared with minoxidil (MNXD) on the growth of human hair dermal papilla cells (HDPCs). Cell viability was measured by MTT assay and cell proliferation was confirmed by cell cycle analysis from flow cytometry in HDPCs. Also, we monitored the safe concentration range through MTT assay. And protein expression of hair growth-related genes (insulin-like growth factor 1 (IGF-1), Wnt3a, Protein kinase B (Akt), Extracellular signal-regulated kinase (Erk)) was monitored by western blot. Results : On cell cycle analysis, the G2/M phase was higher than that of the DW group in AR ethanol extract group at 0.05 and 0.1 mg/㎖. All protein expression levels of HDPCs were increased in AR ethanol extract groups and the MNXD group, compared to the DW group, respectively. Conclusion : As mentioned above, AR extract increased cell proliferation and the protein expression of IGF-1, Wnt3a, Akt, Erk in HDPCs. These results suggest that AR ethanol extract has promoted hair growth and it might be potential hair growth supplement.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6417-6421
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• 2015

Viscum album var (VAV) also known as mistletoe, has long been categorized as a traditional herbal medicine in Asia. In addition to its immunomodulating activities, mistletoe has also been used in the treatment of chronic hepatic disorders in China and Korea. There are numerous reports showing that VAV possesses anti-cancer effects, however influence on human hepatocarcinoma has never been elucidated. In the present study, hot water extracts of VAV was evaluated for its potential anti-cancer effect in vitro. SK-Hep1 cells were treated with VAV (50-400ug/ml) for both 24 and 48 hours then cell viability was measured by cell counting kit-8 (CCK-8). Flow cytometry analysis was used to measure the proportion of SK-Hep1 in the different stages of cell cycle. RT-PCR and Western blot analysis were conducted to measure expression of cell cycle arrest related genes and proteins respectively. VAV dose dependently inhibited the proliferation of SK-Hep1 cells without any cytotoxicity with normal Chang liver cell (CCL-13). Flow cytometry analysis showed that VAV extract inhibited the cell cycle of SK-Hep1 cells via G1 phase arrest. RT-PCR and Western blot analysis both revealed that cyclin dependent kinase 2 (Cdk2) and cyclin D1 gene expression were significantly down regulated while p21 was upregulated dose dependently by VAV treatment. Combined down regulation of Cdk2, Cyclin D1 and up regulation of p21 can result in cell death. These results indicate that VAV showed evidence of anti-cancer activity through G1 phase cell cycle arrest in SK-Hep1 cells.

• 한국미생물·생명공학회지
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• 제32권1호
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• pp.72-77
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• 2004

본 연구진은 토양미생물의 배양액으로부터 cyclin-dependent kinase 저해활성의 Toyocamycin을 분리하였으며 〔16〕, 화학적 전합성을 통하여 활성이 개선된 유도체인 신물질 MCS-5A를 합성하였다〔3〕. 이 MCS-5A를 이용한 항암 기전규명을 위한 연구를 통하여 , human promyelocytic leukemia cell(HL-60)에서 MCS-5A에 의해 cyclin-dependent kinase inhibitor p16$^{INK4A}$ 단백질의 발현증가가 암세포의 세포주기 억제와 동시에 HL-60 cell희 세포사멸을 유도하는 것을 확인하였다(data not shown). 그러나 HL-60 cell의 경우와는 달리 non small cell lung cancer cell(NSCLC)인 A549 cell(p16$^{INK4A}$ 결핍 세포주)에 MCS-5A를 처리할 경우에는 전혀 세포사멸이 유도되지 않았다. 따라서 MCS-5A에 의한 HL-60 cell에서의 세포사멸 유도는 발암억제 유전자인 P16$^{INK4A}$의 세포 내 발현 및 존재 여부에 의해 좌우되는 것으로 판단되었다. 이러한 배경에서 본 연구는 p16$^{INK4A}$.의 기존에 알려진 세포주기 억제를 유발하는 cyclin-dependent kinase inhibitor(CKI)로서의 역할 뿐 아니라, p16$^{INK4A}$ 유전자가 세포사멸을 유도할 수 있다는 새로운 기능을 규명하기 위하여 다음의 연구를 시도하였다. 즉 $p^{INK4A}$ 결핍 세포주인 A549(-p16/+p53)와 H1299(-pl6/-p53) 그리고 p16$^{INK4A}$ 함유 세포주인 HeLa(+p16/+p53)세포에 외부로부터 p16$^{INK4A}$ 유전자를 도입시켜, 각 세포주에서의 세포사멸 유도 여부를 비교하고자 하였다. 우선 wild-type p16$^{INK4A}$ 유전자를 가진 HeLa cell에서 총 RNA를 추출하여, 역전사 반응으로 cDNA를 만들고, PCR을 통해 p16$^{INK4A}$ 유전자를 증폭하였다. pcDNA3.1/His is A vector에 p16$^{INK4A}$ 유전자를 끼워 넣고 competent cell (XL1-Blue)에 형질 전환하여 cloning한 후, p16$^{INK4A}$ clone을 다량으로 추출하였다. 위에 언급한 각각의 cell line에 p16$^{INK4A}$유전자를 농도(0, 1, 5, 10$\mu\textrm{g}$)별로 transfection 시킨 후, p16 단백질을 일정 시간 동안(12시간) 발현시킨 뒤, TUNEL등의 분석을 통해 세포사멸이 유도되는지를 확인하였으며, 또한 Western blot 분석을 통하여 p16단백질과 세포사멸 유도 인자인 caspase 3의 발+현 양상을 확인하였다. 연구 결과, Western blot을 통해 transfection시킨 p16/INK4A/유전자의 농도에 따라 각각의 cell line에서 Pro-caspase 3의 감소함을 관찰할 수 있었고, TUNEL분석을 통해 A549및 HeLa cell에서 세포사멸이 유도됨을 확인할 수 있었다 특히 A549(-p16/+p53)와 HeLa cell(+p16/+p53)에서는 TUNEL 분석 및 Western blot을 통한 pro-caspase 3의 caspase 3로의 전환 등을 통해 세포사멸이 발생하였음을 확연하게 확인할 수 있었으나, 반면 H1299(-pl6/-p53) cell에서는 단지 Western blot을 통한 pro-caspase 3의 활성화만을 통해 간접적으로 세포사멸을 확인 할 수 있었다. 또한 p53이 결핍된 H1299(-pl6/-p53)세포주에서의 $^{INK4A}$ 에 의한 세포사멸 유도는 p53 비의존적으로 작용한다는 사실을 확인할 수 있었다. 결론적으로 발암억제 유전자인 $^{INK4A}$ 는 CKI로서의 기능뿐 아니라, 세포사별 유도와도 밀접하게 관련되어 있으며, 이 기능은 발암 억제 유전자인 p53과는 독립적으로 작용한다는 사실을 확인하였다. 세포사멸 유도 기전연구에서 $p16^{INK4A}$ 가 세포사멸을 유도하는 기전에 대해서는 아직 명확하게 밝혀진 바는 없으며, 현재 본 연구실에서 다양한 실험을 통해 연구가 진행 중이다.

• Nutrition Research and Practice
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• 제14권5호
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• pp.478-489
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• 2020

BACKGROUND/OBJECTIVES: Morusin, a marker component of Morus alba L., possesses anti-cancer activity. The objective of this study was to determine autophagy-inducing effect of morusin in non-small cell lung cancer (NSCLC) cells and investigate the underlying mechanism. SUBJECTS/METHODS: Autophagy induction and the expression of autophagy-related proteins were analyzed by LC3 immunofluorescence and western blot, respectively. The role of autophagy and AMP-activated protein kinase (AMPK) was determined by treating NSCLC cells with bafilomycin A1, an autophagy inhibitor, and compound C, an AMPK inhibitor. Cytotoxicity and apoptosis induction were determined by MTT assay, trypan blue exclusion assay, annexin V-propidium iodide (PI) double staining assay, and cell cycle analysis. RESULTS: Morusin increased the formation of LC3 puncta in the cytoplasm and upregulated the expression of autophagy-related 5 (Atg5), Atg12, beclin-1, and LC3II in NSCLC cells, demonstrating that morusin could induce autophagy. Treatment with bafilomycin A1 markedly reduced cell viability but increased proportions of sub-G1 phase cells and annexin V-positive cells in H460 cells. These results indicate that morusin can trigger autophagy in NSCLC cells as a defense mechanism against morusin-induced apoptosis. Furthermore, we found that AMPK and its downstream acetyl-CoA carboxylase (ACC) were phosphorylated, while mammalian target of rapamycin (mTOR) and its downstream p70S6 kinase (p70S6K) were dephosphorylated by morusin. Morusin-induced apoptosis was significantly increased by treatment with compound C in H460 cells. These results suggest that morusin-induced AMPK activation could protect NSCLC cells from apoptosis probably by inducing autophagy. CONCLUSIONS: Our findings suggest that combination treatment with morusin and autophagy inhibitor or AMPK inhibitor might enhance the clinical efficacy of morusin for NSCLC.

• 대한임상검사과학회지
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• 제38권1호
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• pp.72-81
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• 2006

Wee1 is a kinase regulator of the M-phase promoting factor (MPF; a complex of cdc2 and cyclin B1). The present study was undertaken to determine the role(s) of wee1 in the early stages of mouse ovarian follicles. The expression of wee1 and the correlated cell-cycle components, namely cdc2, cyclin B1, and cdc25C, were evaluated by immunohistochemistry. In addition, the expression of Tyr15-phosphorylated cdc2 (cdc2-p) was also examined to determine whether wee1 kinase phosphorylates cdc2 existed. Each component except cdc25C was found cytoplasmic in the oocytes at all stages of follicles, while cdc25C was not detected in primordial follicles. It was found primarily in ovarian somatic cells and to a small extent in granulosa cells of the growing follicles. To further confirm the expression of cell-cycle components in the primordial follicular oocytes, day1 ovaries were enzymatically and mechanically dissociated, then oocytes were isolated from somatic including pre-granulosa cells, and we confirmed that cdc2-p was expressed in oocytes of primordial follicles. From the results of the present study, we concluded wee1, without the counteracting cdc25C, would cause meiotic arrest of oocytes by the inhibitory phosphorylation of cdc2. The expression of all these proteins in the granulosa cells of growing follicles may regulate their mitosis concurrently with the growth of oocytes and follicles.