• Journal of Acupuncture Research
• /
• v.24 no.2
• /
• pp.51-61
• /
• 2007

목적 : 이 연구는 봉독의 주요 성분인 낮은 농도의 melittin이 in vitro에서 세포자멸사 관련 단백질과 전립 선암세포 PC-3 증식 관련 수용체의 발현 조절을 통하여 세포자멸사(Apoptosis)를 유도하는지 in vivo에서 또한 전립선 암세포주인 PC-3 세포의 성장을 억제하는지 살펴보고자 하였다. 방법 : Melittin을 처리한 후 전립선암세포 PC-3의 성장억제를 관찰하기 위해 WST-l assay와 morphology analysis를 시행하였고， 세포자멸사 관련 MAP kinase 계열의 대표인 ERK1/2과 전립선암세포 증식관련 수용체인 PDGF-BB receptor ${\beta}$의 활성 변화 관찰에는 western blot analysis 및 Immunofluorescence Staining , Confocal immunocytochemistry를 시행하였으며， 전립암세포의 종양형성에는 흉선을 제거한 쥐에 Tumorigenecity study를 시행하였다. 결과 : 1. PC-3 세포에서 Melittin 처리 후 세포증식이 억제되었고 세포의 형태는 세포자멸사의 특징을 나타내었다. 2. PC-3 세포에서 Melittin 처리 후 ERKl/2과 PDGF-BB receptor ${\beta}$의 활성이 억제되었다. 3. PC-3 세포에서 Melittin과 AG1296을 함께 투여시 PDGF-BB receptor ${\beta}$ 활성억제의 상승효과가 나타났다. 4. 흉선 제거 후 전립선암세포주를 이식한 쥐에서 Melittin을 피내로 주입한 결과 전립선암의 크기와 무게가 유의하게 감소하였다. 결론 : 이상의 결과는 Melittin이 ERKl/2과 PDGF BB receptor ${\beta}$의 활성 억제를 통하여 인간 전립선암세포주인 PC-3의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것이며, 이를 재확인한 생처 연구에서의 긍정적인 결과는 향후 Melittin의 전립선암 예방과 치료에 대한 효과적인 치료제 개발에 초석이 될 것으로 기대된다.

• Journal of Life Science
• /
• v.20 no.6
• /
• pp.877-884
• /
• 2010

We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

• Korean Journal of Clinical Laboratory Science
• /
• v.39 no.2
• /
• pp.68-75
• /
• 2007

After reports on regression of cancer in humans and animals infected with microbial pathogens date back more than 100 years, much effort has been spent over the years in developing a wild type or attenuated bacterial and purified bacterial proteins for the treatment of cancer. Pseudomonas aeruginosa exotoxin A (ETA) is known to inhibit cell growth and trigger significant cell death in various cancer cells. Although ETA induces apoptosis of cancer cells, its exact mechanism of action is not known yet. Four different assays were performed in this study: morphological assessment of apoptotic cells, cell cytotoxity, annexin-V binding assay, and cell cycle analysis. The proliferation and survival of the K-562 cells treated with ETA were decreased in a dose dependent manner. In addition, the apoptotic body of K-562 cells was induced by ETA treatment in a dose dependent manner. The ETA-induced apoptosis was confirmed by annexin-V binding assay. Flow cytometric analysis was examined to ascertain whether ETA could arrest the cell cycle at the sub-G1 phase. Our results suggest that P. aeruginosa ETA inhibits cell growth and induces apoptosis in human leukemia K-562 cells.

• Journal of Periodontal and Implant Science
• /
• v.24 no.3
• /
• pp.513-528
• /
• 1994

배양한 치은섬유아세포와 3T3 세포에서, glycyrrhetinic acid가 cyclosporin A를 처리 세포의 활성에 미치는 영향을 알아보기 위해서 MTT 방법을 이용하여 측정하였다. Cyclosporin A는 $1{\sim}10ng/ml$의 농도에서 인체 치은 섬유아세포와 마우스 3T3 세포 활성을 증가시켰으며, glycyrrhetinic acid는 농도와 비례하게 마우스 3T3 세포의 성장을 억제시켰다. 특히 $25{\mu}g/ml$ 이상의 농도에서는 현저하게 억제시킴을 관찰할 수 있었다. 반면, 인체 치은 섬유아세포에서는 $50{\mu}g/ml$ 이상의 농도에서 조차도 성장을 억제 시키지 않았다. 또한, 마우스 3T3와 인체 치은 섬유아세포에서 일정한 cyclosporin A 농도에 $1-100{\mu}g/ml$의 다양한 농도로 glycyrrhetinic acid를 첨가하였을 때, cyclosporin A 단독으로 처리한 세포에 비하여 유의성 있게 세포의 활성을 억제시켰으며, 특히, $100{\mu}g/ml$의 glycyrrhetinic acid를 첨가하였을 경우 세포활성을 현저하게 억제시켰다. 3T3 세포에 cyclosporin A와 $50{\mu}g/ml$의 glycyrrhetinic acid를 함께 처리한 경우 glycyrrhetinic acid단독처리한 군에 비하여 세포활성에 첨가 효과를 보였으며, 인체 치은 섬유아세포 에서는 같이 처리한 경우 glycyrrhetinic acid단독 처리에 비하여 뚜렷한 억제 효과를 나타내었다. 이러한 상승효과는 1ng/ml 의 cyclosporin A와 $50{\mu}g/ml$ glycyrrhetinic acid를 같이 처리한 군에서 가장 뚜렷하게 관찰되었다. 이러한 결과로, 인체 치은 섬유아세포에 cyclosporin A와 glycyrrhetinic acid를 동에 처리하였을 경우 세포의 활성에 상승 효과가 있음을 알 수 있다.

• Environmental Analysis Health and Toxicology
• /
• v.19 no.3
• /
• pp.235-240
• /
• 2004

본 연구는 비만세포의 일종인 RBL-2H3 세포주에 대한 hexamethylene bisacetamide (HMBA)의 세포독성 유도 기전이 telomerase의 활성과 관계가 있다는 것에 대한 보고이다. RBL-2H3 세포주에 5mM HMBA를 처리시 그 세포주의 세포성장이 억제 되었다. 기전연구로서 HMBA는 telomerase활성을 억제 하였으며 RT-PCR에 의한 결과 telomerase mRNA발현의 억제에 의한 것이었다. Madl은 telomerase의 발현을 억제하는 억제인자로 잘 알려져 왔으며 이를 확인해본 결과 Madl은 발현은 유의성있게 증가 되었다. 이를 종합해보면, HMBA에 의한 RBL-2H3 세포주에 대한 세포독성은 Madl 발현의 증가에 따른 telomerase발현의 억제에 기인하는 것으로 밝혀졌다.

• Journal of Life Science
• /
• v.16 no.4
• /
• pp.659-663
• /
• 2006

The extracts of Plocamium telfairiae using several solvents with different polarities were prepared and their growth inhibitory effects were examined on the human cancer cells. We investigated the cytotoxic effects of P. telfairiae extracts on HT-29 cells by the MTT reduction assay and examining the morphological change under the inverted microscope. Among three extracts, the methanol extract showed the strongest inhibitory effect on the growth of HT-29 cells. The methanol extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and aqueous for purifying crude methanol extract. The n-hexane layer among the fractioned layers showed remarkable inhibitory activity on the growth of HT-29 cells. Moreover n-hexane layer showed the notable growth inhibition effects with a dose-dependent manner against SW620, HeLa, and MCF-7 cells as well as HT-29 cells. These results indicated that P. telfairiae extracts may be contained bioactive materials with inhibitory effect on the growth of human cancer cells.

• Journal of Life Science
• /
• v.22 no.9
• /
• pp.1224-1230
• /
• 2012

Cancer chemotherapy drugs command a large share of the market, and the development of new therapeutics with high efficacy and specificity is an active area of study. Recently, the development of cancer therapeutics from natural products targeting angiogenesis has drawn attention due to conventional chemotherapeutics showing serious side effects and resistance in cancer cells. In this study, we investigated the pharmacological efficacy of Gomisin A, an active ingredient of Schizandra chinensis baillon, on tumor growth and metastasis. Administration of Gomisin A at 10 and 100 ${\mu}g/ml$ reduced tumor growth in vivo by $80.5{\pm}8.1%$ and $96.2{\pm}2%$, respectively, compared with positive tumor controls. Treatment of Gomisin A in normal and various tumor cell lines did not exert significant toxicity. Mice treated with Gomisin A at a concentration of 10 and 100 ${\mu}g$/head showed a significant reduction in tumor-induced angiogenesis of $151{\pm}16.9%$ and $98.5{\pm}29.5%$, respectively. Furthermore, tumor metastasis analysis revealed that the administration of Gomisin A at a concentration of 10 and 100 ${\mu}g$/head inhibited tumor metastasis by $13.5{\pm}8.56%$ and $58.3{\pm}9.12%$, respectively. In addition, Gomisin A significantly decreased cell adhesion of the B16BL6 cells to the extracellular matrix. These results demonstrate that Gomisin A inhibits tumor growth via suppression of angiogenesis and tumor metastasis inhibition, without cellular toxicity. The pharmacological efficacy of Gomisin A suggests that it may be a potential candidate for the development of cancer drugs.

• Journal of Acupuncture Research
• /
• v.28 no.3
• /
• pp.111-119
• /
• 2011

목적 : 이 연구는 봉독이 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도하고 전립선 암세포주인 DU-145 세포의 성장을 억제하는지를 확인하고 해당 기전을 살펴보고자 하였다. 방법 : 봉독을 처리한 후 DU-145의 세포자멸사를 관찰하기 위해 TUNEL staining assay를 시행하였으며, 세포자멸사 조절단백질의 변동 관찰에는 western blot analysis를 시행하였다. 결과 : DU-145 세포에 봉독을 처리한 후, 세포자멸사의 유발, 세포자멸사 관련 단백질의 발현에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. DU-145 세포에서 봉독을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었다. 2. 세포자멸사 관련 단백질 중 분리된 pro-apoptotic proteins인 PARP, caspase-3, caspase-9은 유의한 증가를 나타내었다. 3. 세포자멸사 관련 단백질 중 분리된 anti-apoptotic proteins인 Bcl-2, p-AKT, XIAP, cIAP2는 유의한 감소를, MMP2, MMP13은 유의한 증가를 나타내었다. 결론 : 이상의 결과는 봉독이 인간 전립선 암세포주인 DU-145의 세포자멸사를 유발함으로써 전립선암세포 증식억제 효과가 있음을 입증한 것으로 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

• Korean Journal of Food Science and Technology
• /
• v.36 no.1
• /
• pp.141-146
• /
• 2004

Fungal products indirectly mediate anti-tumor effects in vitro and in vivo. To investigate whether Lentinus edodes might possess direct anti-tumor substance, L. edodes was extracted and tested on human papillomavirus (HPV) 16 oncogenes-associated animal tumor cells (TC-1) and in an animal tumor model. Only water extract displayed direct anti-proliferative effects in TC-1 tumor cells in vitro. This inhibition was dose-dependent, and inhibitory concentration ($IC_{50}$) was $800\;{\mu}g/mL$. Fungal extracts also showed growth inhibition to human cervical cancer cells (CaSki and HeLa) similarly to TC-1 tumor cells. When fungal extracts were added at a high dose (1.5 mg/mL), cell growth was inhibited within 6 hr following extract treatment. Cell growth inhibition was blocked by heat treatment, but not by low pH, which is indicative of heat sensitivity of this anti-proliferative substance. Cell growth suppression was mediated by apoptosis, as determined by Annexin V and propidium iodide staining. When challenged with TC-1 cells, direct intratumoral injection of fungal extracts resulted in some positive effect on tumor growth inhibition, as compared to oral delivery. Results suggest that heat labile substance of L. edodes suppresses growth of HPV oncogenes-associated tumor cells through apoptosis.

• The Korean Journal of Zoology
• /
• v.33 no.1
• /
• pp.45-52
• /
• 1990

In order to determine whether maturation inhibiting activity(MIA) in the cytoplasm of growing follicular oocytes would suppress the cleavage of the embryonal cells, the growing oocytes were fused with the 2 or 4 cell blastomeres and then examined for the nuclear phase of the fused giant cells 24 hr after culture. A significant number of the giant cells(60%) composed of growing mouse oocyte and 2 cell mouse blastomere(1/2) in interphase has contained 2 nuclei 24 hr after culture and most of the giant cells (90%) composed of the growing oocyte and 4 cell blastomere(1/4) also contained 2 nuclei after culture. The unfused blastomeres or the isolated blastomeres cultured without fusion treatment cleaved one cell cycle under the same culture condition. In contrast, the nucleus of the growing oocytes was disintegrated and the chromosome condensed when fused with 2 cell blastomere in mitosis. The growing rat oocytes also suppressed the nuclear disintegration of the mouse embryonal cells during culture. The data presented here showed that MIA in the growing mammalian oocyte inhibited the cleavage of the embryonal cells in interphase stage, but not in milosis stage.