• 한국수정란이식학회지
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• 제24권3호
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• pp.183-187
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• 2009

In this study, we aimed to determine whether the evaluated markers of cell death could be found at particular developmental stages of normal porcine in vitro fertilization (IVF) embryos. We investigated the characteristics of spontaneous and induced apoptosis during preimplantation development stages of porcine IVF embryos. In experiment 1, to induce apoptosis of porcine IVF embryos, porcine IVF embryos at 22h post insemination were treated at different concentration of actinomycin D (0, 5, 50 and 500 ng/ml in NCSU medium). Treated embryos were incubated at $39^{\circ}C$ in 5% $CO_2$, 5% $O_2$ for 8h, and then washed to NCSU medium and incubated until blastocyst (BL) stage. We examined cleavage rate at 2days and BL development rate at 7days after in vitro culture. A significantly lower rate of cleavage was found in the 500 ng/ml group compared to others (500 ng/ml vs. 0, 5, 50 ng/ml; 27.8 % vs. 50.0%, 41.2%, 35.9%), and BL formation rate in 500 ng/ml was lower than that of others (500 ng/ml vs. 0, 5, 50 ng/ml; 8.0% vs. 12.6%, 11.2%, 12.6%). In experiment 2, to evaluate apoptotic cells, we conducted TUNEL assay based on morphological assessment of nuclei and on detection of specific DNA degradation under fluorescence microscope. This result showed that apoptosis is a normal event during preimplantation development in control group (0 ng/ml actinomycin D). A high number of BL derived control group contained at least one apoptotic cell. Actinomycin D treated BLs responded to the presence of apoptotic inductor by significant decrease in the average number of blastomeres and increase in the incidence of apoptotic cell death. In 500 ng/ml group, the incidence of apoptosis increased at 4-cell stage and later. This result suggested that apoptosis is a process of normal embryonic development and actinomycin D is useful tool for the apoptosis study of porcine preimplantation embryos.

• 대한한방부인과학회지
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• 제21권3호
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• pp.18-33
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• 2008

Purpose: The research is to investigate the effect of TFE on apoptosis of human-derived breast cancer cells, to find out the relationship with apoptosis. Methods: Human-derived breast adenocarcinoma cell line, MCF-7 cells were treated by TFE with various concentration. The inducement effect of TFE on cell apoptosis was observed with MTT assay and the relationship between the treatment and apoptosis was investigated with FACS analysis, TUNEL assay and DNA laddering assay and the change in the protein levels of PARP and caspase-3 activities were also observed. The release of cytochrome-c was observed to find out the pathway of apoptosis induced by TFE. Results: The cell apoptosis was significantly induced in MCF-7 cells treated with TFE in concentration-dependent and time-dependent manner. It was verified by FACS analysis, TUNEL assay, DNA laddering assay that cell-death was caused not by necrosis but by apoptosis. The activity of PARP and caspase were increased concentration-dependently. The release of cytocrome-c was decreased in proportion to the concentration of the fruit extract. It therefore demonstrated that mitochondria were involved in apoptosis induced by TFE. The appearance of Bcl-2 protein was decreased concentration-dependently. Conclusion: The treatment by TFE induced apoptosis of human breast adenocarcinoma cell line, MCF-7. It seems likely that cell-death was caused by apoptosis and mitochondria were involved in it. The mechanism of protein change causing apoptosis seems related to the inhibition of Bcl-2 protein, the promotion of inversion from cytochrome-c into cytosol, the activation of caspase and the promotion of PARP cleavage.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1027-1033
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• 2007

Paclitaxel이 암세포에서 세포예정사를 유발할지라도, 아직 정확한 기전은 잘 알려져 있지 않다. 이에 본 연구에서는 HeLa $S_{3}$ 자궁암세포에서의 paclitaxel이 어떠한 영향을 미치는지 알아보고자 한다. 그리하여 방법으로는 세포독성검사, apoptotic cells의 형태학적 변화(DAPI 염색 ), western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 paclitaxel은 HeLa $S_{3}$ 세포에서 세포독성을 보이며 특히 paclitaxel의 $IC_{50}$ 값은 약 1 ${\mu}M$이며, paclitaxel 처리한 HeLa $S_{3}$ 세포에서 형태학적 변화(분절화)를 관찰하였고, flow cytometric 분석에서는 G2/M기가 차단되어 paclitaxel은 세포주기 특히 Sub-$G_{1}$기를 조절함을 알 수 있다. 그리고 Paclitaxel을 처리한 HeLa $S_{3}$ 세포에서는 PARP cleavage를 유발하였고 Bc1-2의 감소와도 관련되었다.

• 생명과학회지
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• 제31권4호
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• pp.430-441
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• 2021

세스퀴테르펜 락톤(Sesquiterpene lactones; STL) 화합물은 테르페노이드의 일종으로 주로 국화과에서 발견이 되고 강한 세포 독성을 나타내는 생리학적 특성을 지니고 있다. 이러한 세스퀴테르펜 락톤은 강한 세포 독성으로 인해 연구가 미미하였으나, 최근 화학적 변형을 통해 독성이 적은 형태로 합성하여 새로운 의약품 개발로서의 연구가 활발히 진행되고 있다. 세스퀴테르펜 락톤 화합물인 artemisinin 및 mipsagargin 화합물은 현재 말라리아 및 종양성장에 대한 약물로 사용되고 있다. 또한 항산화, 간보호, 항바이러스, 항균, 항종양 및 항노화 등의 생리활성 효능이 보고되어 있으며, 종양세포에서 자멸사를 유도하여 항암제로서의 연구가 진행되고 있다. 본 연구에서는 세스퀴테르펜 락톤 화합물인 artemisinin, costunolide, thapsigargin, arglabin, parthenolide, alantolactone, cynaropicrin, helenalin, 및 santonin의 생리활성 효능에 대한 연구 동향을 검토하고자 한다.

• 대한한방부인과학회지
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• 제18권1호
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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.

• Clinical and Experimental Reproductive Medicine
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• 제35권1호
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• pp.61-67
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• 2008

목 적: Glucocorticoid가 황체용해에 미치는 효과의 기전의 일부를 면역 조직학적 연구를 통해 알아보고자 하였다. 연구방법: 8주령의 Rat 암컷 30마리 대조군, 덱사메타손 0.1 mg 투여군 및 덱사메타손 1mg 투여군의 3군으로 나누어 황체용해의 차이와 대식세포의 침투여부를 비교하기 위하여 광학현미경을 이용하여 계수하였다. 결 과: TUNEL 면역염색 결과 덱사메타손 0.1 mg 투여군과 1 mg 투여군 모두에서 대조군에 비해 황체용해의 비율이 현저히 감소하였고, ED1 면역염색 결과 덱사메타손 1 mg 투여군에서 대식세포의 황체 내 침투가 현저히 억제된 것으로 나타났다. ED1 면역염색 결과 덱사메타손 1 mg 투여군에서 대식세포의 면역반응성이 대조군에 비해 감소되어 나타났다. 결 론: 덱사메타손은 직접 황체세포에 작용하여 세포자멸사를 억제한다. 또한 대량 투여했을 때는 대식세포의 황체 내 침투를 억제하거나 황체 내에 이미 존재하는 대식세포의 활성화를 억제함으로써 황체용해를 억제한다.

• 한국자원식물학회지
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• 제31권2호
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• pp.102-108
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• 2018

본 연구는 꼭두서니의 뿌리인 천초근의 물 추출물과 에탄올 추출물을 이용하여 대장암 세포에 대한 암세포 성장 억제 및 사멸효과가 있는지 알아보고자 수행하였다. ERA(천초근 에탄올 추출물)은 폴리페놀($45.77{\pm}2.03mg/g$)과 플라보노이드($22.82{\pm}1.33mg/g$)를 함유하고 있었으며, $H_2O_2$에 의해 증가된 ROS(reactive oxygen species)를 억제하는 효과를 나타냈지만 WRA(천초근 물 추출물)은 효과가 없었다. 또한 ERA는 $500{\mu}g/m{\ell}$의 농도로 대장암 세포주(HCT-116)에 처리했을 때 세포사멸을 유도할 뿐만 아니라 caspase-3 단백질 활성화, DNA fragmentation 및 apoptotic cell death를 일으키는 것으로 확인되었다. 이는 ERA가 HCT-116 세포주에 대해 apoptosis(세포자멸사)를 통해 항암효과를 나타내는 것으로 생각되지만 다른 연구결과들과 비교하였을 때 농도 대비 효능이 미미하다. 따라서 천초근 에탄올 추출물에 대장암 세포의 성장을 억제하는 유효성분을 분석하여 그 효능을 탐색하는 추가실험이 필요할 것으로 생각된다.

• 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.

• 대한한방부인과학회지
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• 제19권3호
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• pp.135-149
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Millettia Reticulatas on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Millettia Reticulatas and investigated cell death rate by MTS assay. Furthermore, flow cytometry analyis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Millettia Reticulatas was increased in a concentration proportional. 2) The result of flow cytometry analysis. subG1 phase arrest related3 cell apoptosis was investigated 23.49% in uterine leiomyoma cell treated Millettia Reticulatas and showed the fession of proportional concentration. 3) The gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing concentration but cyclin E was none exchanged. 4) The character of apoptosis, DNA fragmentation was significantly observed the fession of proportional concentration. 5) The expression of pro-caspase3 and PARP were decreased dependent on treatment concentration. Conclusion : This study showed that Millettia Reticulatas have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis. The apoptotic mechanism was observed that the gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing treatment concentration, induced G1 phase arrest and finally cell death was occurred. The decreased expression of pro-caspase 3 and PARP were noted that apoptosis was related with caspase pathway.

• 대한한방부인과학회지
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• 제27권2호
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• pp.12-22
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• 2014

Objectives: This study aimed to evaluate the effects of Cuscutae Semen water extract (CS) on MCF-7 human breast cancer cells. Methods: To clarify the results, we cultivated MCF-7 cells in cell culture plates. And then we extracted each of $100{\mu}g/ml$, $300{\mu}g/ml$, $600{\mu}g/ml$ CS, gave it to MCF-7 cell. After these process we performed MTT assay to elucidate the ability of apoptosis. The result of mRNA was analyzed by RT-PCR. Results: Each of concentrated extracts CS decreased the survival rate of MCF-7 cells. CS decreased Bcl-2 which is known as a blocking cell apoptosis. Bax, caspase-3, P21 and RIP-1 that accelerate apoptogenic activity factors increased by CS. CS did not change the condition of caspase-8, caspase-9, P53 factors on MCF-7 cells. Furthermore caspase-8, caspase-9, P53 factors on MCF-7 cells does not make it more active but turn it on. Conclusions: According to the above results, we could suggest that CS can occur the apoptosis on MCF-7 cells.