• Journal of Applied Biological Chemistry
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• 제63권2호
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• pp.147-152
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• 2020

Farrerol은 중국에서 거담제로 사용되어온 전통 한약제로 사용된 산진달래(만산홍, Rhododendron dauricum L.)에서 유래된 플라바논이다. Farrerol은 항산화, 항염증 및 항균 작용을 포함한 다양한 생리 활성이 보고되었다. 하지만 farrerol의 MCF-7에 대한 항암 작용은 아직 보고된 바가 없다. 본 연구에서 인간 유방암 MCF-7 세포에 대한 farrerol의 처리가 세포증식을 억제하고 apoptosis를 유도함을 입증하였다. MCF-7 세포에 ferrerol을 48시간 동안 처리했을 때, 이는 통계적으로 유의한 세포증식 효과를 나타냈으며 이의 IC₅₀ 값은 145.04±1.4 μM임을 확인하였다. 또한, farrerol이 세포사멸을 유도함을 TUNEL assay와 FACS를 이용한 Annexin V/PI 염색을 통해 검증하였다. 이러한 항암 효능의 작용기전으로써, farrerol의 처리가 BAX/Bcl-2 및 Caspase-3활성화와 PARP 분절화를 증가시켜 세포자살을 촉진한다는 것을 확인하였다. 결론적으로, 본 연구의 결과는 farrerol이 apopotosis 관련 단백질의 활성 및 발현조절을 통해 MCF-7 유방암세포에 대한 항암 효능을 갖는다는 것을 보여주고 있다.

• Journal of Nutrition and Health
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• 제51권6호
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• pp.498-506
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• 2018

본 연구는 베타세포에서 라이코펜의 항사멸 효과와 그 기전에 대해 조사하기 위해 실시하였다. 라이코펜에 의한 베타세포독성을 조사하기 위해 다양한 농도 (0.1, 1, 10 nM)로 처리하였을 경우, 저농도에서 세포독성이 나타나지 않음을 관찰하였다. 선택한 농도를 사이토카인 혼합물과 함께 처리하였을 경우, 세포 생존율이 증가하는 것을 관찰하였고, 세포사멸 유도 단백질인 Bax의 발현양은 감소하고, 세포사멸억제 단백질인 Bcl-2 발현양은 증가하는 것을 관찰하였다. 또한 사이토카인 혼합물에서 증가하였던 세포내 산화스트레스가 라이코펜과 함께 처리하였을 경우 감소되는 것을 관찰하였고 이러한 효과는 항산화 유전자인 GCLC, NQO1, HO-1의 발현양이 증가함으로서 일어난 현상임을 알 수 있었다. 라이코펜은 미토콘드리아의 생성 및 기능과 관련된 유전자의 발현을 증가시키고 사이토카인 혼합물에 의해 감소되었던 세포내 ATP 생성량을 증가시켰다. 이러한 결과는 라이코펜의 항산화효과와 미토콘드리아 기능 개선 효과가 사이토카인에 의한 베타세포 사멸을 억제하는 기전 중의 하나로 작용할 수 있음을 의미한다. 향후 라이코펜이 베타세포를 타겟으로 하는 제 2형 당뇨 치료의 기능성 소재로 개발될 가능성이 있음을 시사하는 바이다.

• Clinical and Experimental Pediatrics
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• 제51권2호
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• pp.170-180
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• 2008

목 적 : 몇몇 항생제가 저산소성 허혈성 뇌 손상에서 뇌 보호 효과를 가진 것으로 밝혀졌지만 아직까지 그 기전에 대해 잘 알려지지 않고 있다. 최근 아미노글루코사이드 계열의 항생제인 G418(geneticin)이 항고사에 의한 암세포 생존을 증가 시키는 것으로 알려졌으며 본 연구는 G418이 주산기 저산소성 허혈성 뇌손상에서 세포 고사를 억제함으로서 뇌 보호 효과를 나타내는지를 알아보고자 하였다. 방 법 : 임신 18일된 백서의 대뇌피질 신경세포를 배양하여 정상산소 상태군와 저산소 상태군으로 나누고, 두 군을 각각 대조군과 G418 $10{\mu}g/mL$으로 처리한 군으로 나누어서 TUNEL 분석과 caspase 3에 대한 면역조직생화학검사를 하였고, 생후 7일된 신생 백서의 좌측 총경동맥을 결찰 후 절단하고 저산소 상태를 유도하여 G418을 저산소 상태 유도 전 30분과 유도 후 30분에 복강 내로 $0.1{\mu}g/kg$ 투여하고 7일 후에 희생시켜 H&E 염색과 caspase-3에 대해 Western blot과 real-time PCR를 하였다. 결 과 : TENEL 분석상 정상산소 상태군과 저산소 상태군 모두에서 대조군보다 G418 투여군에서 통계학적으로 유의하게 고사세포가 감소되었다(P<0.01). Caspase-3에 대한 면역조직화학검사에서 저산소 손상 전에 G418을 투여한 군에서 손상 후에 투여한 군보다 Caspase-3 발현이 적게 나타났다. 저산소 손상 7일 후에 얻은 신생 백서 뇌의 육안적 관찰과 H & E 염색에서 저산소 상태군의 뇌 용적이 정상산소 상태군의 경우보다 감소된 것을 알 수 있었고, 저산소 손상 전에 G418을 투여한 군에서 손상 후에 투여한 군보다 뇌 조직의 손상이 더 적은 것을 볼 수 있었다. Bax/Bcl-2, caspase-3에 대해 Western blot과 real-time PCR 한 경우에 G418 투여한 군에서 Bax/Bcl-2 비율과 caspase-3 발현이 감소되었다. 결 론 : 주산기 저산소성 허혈성 뇌 손상에서 G418는 뇌 조직의 고사를 억제함으로서 뇌보호 효과를 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1287-1293
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• 2011

The extracellular polysaccharide (EPS) was isolated from mycelial cultures of Laetiporus sulphureus var. miniatus and purified by DEAE cellulose and Sephadex G-50 column chromatography. The purified EPS (EPS-2-1) was composed of only glucose units and its molecular mass was 6.95 kDa. The chemical structure of EPS-2-1 consisted of a main chain containing ($1{\rightarrow}4$)-Glcp units with branches at the C-6 position of the chain carrying-Glcp-($1{\rightarrow}4$)-linked residues. The effect of purified EPS on immunomodulatory genes and proteins of the Bcl-2 family was observed using cultured U937 human leukemia cells. Of note, the levels of Bax and Bad proteins treated with the EPS (4 mg/ml) were approximately 23- and 18-times higher than those in non-treated cells, respectively. These results may suggest that the EPS purified from the mushroom L. sulphureus is associated with the activation of immunomodulatory mediators, Bax and Bad proteins.

• Biomolecules & Therapeutics
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• 제25권2호
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• pp.158-164
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• 2017

Methylglyoxal (MGO) is a highly reactive metabolite of glucose which is known to cause damage and induce apoptosis in endothelial cells. Endothelial cell damage is implicated in the progression of diabetes-associated complications and atherosclerosis. Hypericin, a naphthodianthrone isolated from Hypericum perforatum L. (St. John's Wort), is a potent and selective inhibitor of protein kinase C and is reported to reduce neuropathic pain. In this work, we investigated the protective effect of hypericin on MGO-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Hypericin showed significant anti-apoptotic activity in MGO-treated HUVECs. Pretreatment with hypericin significantly inhibited MGO-induced changes in cell morphology, cell death, and production of intracellular reactive oxygen species. Hypericin prevented MGO-induced apoptosis in HUVECs by increasing Bcl-2 expression and decreasing Bax expression. MGO was found to activate mitogen-activated protein kinases (MAPKs). Pretreatment with hypericin strongly inhibited the activation of MAPKs, including P38, JNK, and ERK1/2. Interestingly, hypericin also inhibited the formation of AGEs. These findings suggest that hypericin may be an effective regulator of MGO-induced apoptosis. In conclusion, hypericin downregulated the formation of AGEs and ameliorated MGO-induced dysfunction in human endothelial cells.

• Integrative Medicine Research
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• 제3권2호
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• pp.67-73
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• 2014

Background: The transcription factor signal transducer and activator of transcription 3 (Stat3)is constitutively activated in many human cancers. It promotes tumor cell proliferation,inhibits apoptosis, induces angiogenesis and metastasis, and suppresses antitumor hostimmune responses. Therefore, Stat3 has emerged as a promising molecular target for cancertherapies. In this study, we evaluated the Stat3-suppressive activity of 38 herbal medicinestraditionally used in Korea.Methods: Medicinal herb extracts in 70% ethanol were screened for their ability to suppressStat3 in the A549 human lung cancer cell line. A Stat3-responsive reporter assay system wasused to detect intracellular Stat3 activity in extract-treated cells, and Western blot analyseswere performed to measure the expression profiles of Stat3-regulated proteins.Results: Fifty percent of the 38 extracts possessed at least mild Stat3-suppressive activities(i.e., activity less than 75% of the vehicle control). Ethanol extracts of Bupleurum falcatumL., Taraxacum officinale Weber, Solanum nigrum L., Ulmus macrocarpa Hance, Euonymus alatusSieb., Artemisia capillaris Thunb., and Saururus chinensis (Lour.) Baill inhibited up to 75% of thevehicle control Stat3 activity level. A549 cells treated with these extracts also had reducedBcl-xL, Survivin, c-Myc, and Mcl-1 expression.Conclusion: Many medicinal herbs traditionally used in Korea contain Stat3 activity-suppressing substances. Because of the therapeutic impact of Stat3 inhibition, these resultscould be useful when developing novel cancer therapeutics from medicinal herbs.

• Advances in nano research
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• 제7권4호
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• pp.241-248
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• 2019

Recent researches demonstrated well promising anticancer activities for antibiotics. Such effects would be significantly increased while nanoparticle based delivery systems were applied. In this study, the goal was aim to improve anticancer and antitoxic effects of Streptomycin by loading on special kind of dendrimer (anionic-linear-globular second generation). In the current study, Size and zeta potential as well as AFM techniques have been used to prove the fact that the loading was performed correctly. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the drug loaded on dendrimer nanoparticle were determined and compared with both of dendrimer alone and free drug with respect to staphylococcus aureus as the test microorganism. The anticancer activity among three groups including Streptomycin, Streptomycin -G2 dendrimer, and control was measured in vitro. In vitro studies showed that G2 anionic linear-globular polyethylene-glycol-based dendrimer, which loaded on Streptomycin was able to significantly improve the treatment efficacy over clinical Streptomycin alone with respect to proliferation assay. Maximal inhibitory concentration (IC50) was calculated to be $257{\mu}g/mL$ for streptomycin alone and $55{\mu}g/mL$ for Streptomycin -G2 dendrimer. In addition, Streptomycin -G2 dendrimer conjugate prevented the growth of MCF-7 cancerous cells in addition to enhance the number of apoptotic and necrotic cells as demonstrated by an annexin V-fluorescein isothiocyanate assay. Streptomycin -G2 dendrimer conjugate was able to increase Bcl-2/Bax ratio in a large scale compared with the control group and Streptomycin alone. Based on results a new drug formulation based nano-particulate was improved against S. aureus with sustained release and enhanced antibacterial activity as well as anticancer activity shown for functional cancer treatment with low side effects.

• Nutrition Research and Practice
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• 제3권3호
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• pp.180-184
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• 2009

The apoptotic effect of bacteria-derived $\beta$-glucan was investigated in human colon cancer cells SNU-C4 using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax, and Caspase-3 genes, and assay of caspase-3 enzyme activity. $\beta$-Glucan of 10, 50, and $100{\mu}g$/mL decreased cell viability in a dose-dependent manner with typical apoptotic characteristics, such as morphological changes of chromatin condensation and apoptotic body formation from TUNEL assay. In addition, $\beta$-glucan ($100{\mu}g$/mL) decreased the expression of Bc1-2 by 0.6 times, whereas the expression of Bax and Caspase-3 were increased by 3.1 and 2.3 times, respectively, compared to untreated control group. Furthermore, the caspase-3 activity in the $\beta$-glucan-treated group was significantly increased compared to those in control group (P < 0.05). Bacterial derived $\beta$-glucan could be used as an effective compound inducing apoptosis in human colon cancer.

• 생명과학회지
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• 제25권8호
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• pp.849-860
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• 2015

본 연구에서는 한약재로 널리 사용되는 건칠, 유근피 및 신석 추출물의 항암 활성을 조사하였다. 생쥐 유래 정상세포(RAW 264.7 대식세포 및 C2C12 근아세포)에서는 건칠, 유근피 및 신석 단독 및 복합 처리에 의하여 유의적인 세포생존율의 억제 현상은 관찰 할 수 없었다. 그리고 건칠, 유근피 및 신석의 복합 처리는 단독 처리군에 비하여 AGS 위암세포의 생존력을 유의적으로 억제하였으나, 폐암(A549), 대장암(HCT116), 간암(Hep3B) 및 방광암(T24) 세포에서는 그 효과가 미비하였다. 아울러 이러한 AGS 위암세포 선택적 생존 억제력은 apoptosis 유도와 밀접한 연관성이 있음을 염색질의 응축 현상, DNA 단편화 및 annexin-V 염색에 의한 flow cytometry 분석을 통하여 확인하였다. 건칠, 유근피 및 신석의 복합 처리는 Fas 및 Fas legand의 발현을 증가시켰으며, XIAP, cIAP-1 및 survivin과 같은 IAP family 단백질과 anti-apoptotic Bcl-xL의 발현은 저하시켰다. 복합 처리는 또한 mitochondrial membrane potential의 손실과 caspases (-3, -8 및 -9)의 활성에 PARP 단백질의 분절화를 유도하였다. 그러나 이러한 복합 처리에 의한 AGS 세포에서 관찰된 세포독성 및 apoptosis 유도 효과는 pan-caspases inhibitor인 z-VAD-fmk의 선처리에 의하여 차단되었다. 이상의 결과는 건칠, 유근피 및 신석의 복합 처리에 의한 AGS 위암세포 선택적 apoptosis 유도가 caspase 의존적으로 일어나고 있음을 보여주는 결과이며, in vivo 모델을 이용한 후속 연구가 진행되어야 할 것이다.

• 대한한방종양학회지
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• 제5권1호
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• pp.47-60
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• 1999

Traditional oriental medicines have been used for treatment of various kinds of human cancers for long times and some of them proven to be effective clinically. However, the pharmacological actions and mechanisms related to cancer treatment are generally unknown. In an effort to clarify the action mechanisms of several oriental medicines used for cancer treatments, we planned this experimental procedures. We selected Cordyceps sinensis (冬蟲下草), Punellae Herba (夏枯草), Rehmanniae Radix (熟地黃), Paeoniae Radix (白芍藥), Oldenlandiae Herba (白花蛇舌草), Partulaceae Herba (馬齒? ), Scdopendra subspinipes mutilans (蜈蚣), Mylabris Phalerara (班蟄), Phellinus igniarius(桑黃), Ganodermae Lignum(靈芝) for evaluation, which have been used for patients of gastric cancers. The twenty grams of medicines were boiled in 100ml of water for 1 hour and filtered with $0.2\;{\mu}m$ pore-sized filter unit to remove insoluble particles. Initially we evaluated the effects oriental medicines on growth inhibition in stomach cancer cells. The gastric cancer cell line, AGS, was cultured in RPMI 1640 supplemented with l0% heat-inactivated fetal bovine serum and treated with $10{\mu}l$ oriental medicines per 1ml of medium up to 48 hours. The specimens were subjected to MTT assay for evaluation of growth inhibition. We found mat Mylabris phalerata (班蟄) markedly suppressed the growth of cancer cells as shown in results. Next, we checked the effects of oriental medicines on cancer on cell cycles and apptosis. Mylabrls phalerata (班蟄) inhibited cell cycle progression of cancer cells a compared with control cells and cells treated with other medicines. In addition, Mylabri phalerata (班蟄) induced apoptosis in 30-40% of stomch cancer cells within 24 hours. Other oriental medicines used for this experiments did not show apoptosis-inducing effects on cancer cells. Finally, we determined the expression levels of genes associated with cell cycle and apoptosis. The expressions of Bcl-2 and bcl-XL were downregulated by the treatment of Mylabris phalerata (班蟄). However, the expression levels of genes related to cell cycles were not altered significantly. In conclusion, we found that Mylabris phalerata (班蟄) has in vivo gowth-inhibiting and apptosis-inducing effects on stomach cancer cells. However, we think that at least animal experiments are necessay for evduations.