• Title/Summary/Keyword: calpain

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Identification of Polymorphisms in CAST Gene Associated with Economic Traits in Hanwoo (Bos taurus coreanae) (한우(Bos taurus coreanae)의 CAST 유전자 내 변이지역 탐색 및 경제형질과의 연관성 분석)

  • Oh, Jae-Don;Lee, Jin-A;Lee, Kun-Woo;Park, Kyung-Do;Cho, Byung-Wook;Jeon, Gwang-Joo;Lee, Hak-Kyo;Kong, Hong-Sik
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1498-1504
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    • 2010
  • A number of studies have shown that the calpain system is important in normal skeletal muscle growth. An increased rate of skeletal muscle growth can result from a decreased rate of muscle protein degradation, and this is associated with a decrease in activity of the calpain system, due principally to a large increase in calpastatin (CAST) activity. The CAST gene, mapped to BTA 7, is considered a candidate gene for beef tenderness and muscle growth. The present study used comparative sequencing of five novel polymorphisms located within exon 20 and 22 of the bovine CAST gene in Hanwoo: exon20- 109737G/A, 109749T/C, 109823T/C, exon22- 116151G/A, intron- 109926G/A. The association of the CAST SNPs with economic traits was studied. The 109926G/A showed a significant effect only on the longissimus muscle area (LMA, p<0.05) in Hanwoo. 109926G/A with the genotype GG had a significantly higher effect on LMA (75.35) than the genotype AA (69.6, p<0.05). Also, the 116151G/A showed a significant effect only on weight at 18 months (W18, p<0.05). 116151G/A with the genotype GG had a significantly higher effect on W18 (428.54) than the genotype AA (408.87, p<0.05).

Bee Venom Inhibits Prostate Cancer Growth in LNCaP Xenografts via Apoptosis (Bee venom의 세포자멸사를 통한 전립선 암세포의 성장 및 LNCaP의 이종이식에 미치는 영향)

  • Yang, Chang-Yeol;Song, Ho-Sueb
    • Journal of Pharmacopuncture
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    • v.13 no.1
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    • pp.15-35
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    • 2010
  • 연구목적 : 이 연구는 봉약침의 봉독이 NF-${\kappa}B$ 활성억제와 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 발현을 통하여 세포자멸사를 유도하고, 전립선 암세포를 이식한 쥐에서의 세포자멸사 유도 효과를 확인함으로써, 봉약침의 봉독이 생체 내에서도 세포자멸사를 유도하여 전립선암에 효과를 나타냄을 확인하고자 하였다. 실험방법 : 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절 단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였다. 결과 : 1. DAPI, TUNEL staining assay 결과 봉독 및 melittin을 처리한 LNCaP 세포 모두에서 세포자멸사 유도율이 유의한 증가를 나타내었다. 2. LNCaP 세포에 봉독이나 melittin을 처리한 결과, 안드로겐 수용체 조절 단백질 중 p-Akt, COX-2, calpain은 봉독과 melittin 모두에서 유의한 감소를 나타내었고, Akt는 melittin에서 유의한 감소를 나타냈으며, 봉독에서 증가하는 경향을 보였고, MMP-9은 증가하였다. 3. 생체 내에서의 봉독의 항암효과를 확인하기 위해 전립선암세포가 이식된 쥐에 봉독을 처리한 후 암세포의 부피와 무게, 쥐의 체중을 측정한 결과, 봉독을 처리한 군에서 암 세포 부피비율 및 무게는 감소하였고, 쥐의 체중은 증가하였다. 4. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, NF-${\kappa}B$ 활성에서 유의한 감소를 나타내었다. 5. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, 세포자멸사 조절 단백질 중 Bax/Bcl-2, p53, caspase-3, caspase-9, calpain은 유의한 증가를, COX-2는 유의한 감소를 나타냈으며, MMP-9는 증가를 나타내었다. 결론 : 이상의 결과는 봉독이 시험관 내에서 뿐만 아니라 생체 내에서도 NF-${\kappa}B$의 활성을 억제하고 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 조절을 통하여 인간 전립선암 세포주인 LNCaP의 세포자멸사를 유도함으로써 전립선암 세포 증식억제 효과 및 호르몬 비의존적인 전립선암으로의 전이를 지연시키는 경향이 있을 것으로 사료되고, 봉독이 전립선암의 예방과 치료에 효과적으로 활용될 수 있을 것으로 기대된다.

Lovastatin Induces Apoptotic Cell Death by Activation of Intracellular Ca2+ Signal in HepG2 Human Hepatoma Cells

  • Lee, Yong-Soo
    • Biomolecules & Therapeutics
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    • v.15 no.3
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    • pp.137-144
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    • 2007
  • Although lovastatin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMGCoA) reductase, has been shown to have anti-cancer actions, the effect on human hepatoma cells was not investigated. Moreover, the exact mechanism of this action is not fully understood. In this study we investigated the mechanism by which lovastatin induces apoptosis using HepG2 human hepatoblastoma cells. Lovastatin induced apoptotic cell death in a dose-dependent manner in the cells, assessed by the flow cytometric analysis. Treatment with mevalonic acid, a precursor of cholesterol, did not significantly suppress the lovastatin-induced apoptosis. Lovastatin induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the lovastatin-induced intracellular $Ca^{2+}$ increase and apoptosis, whereas intracellular $Ca^{2+}$ reduction with BAPTA/AM and intracellular $Ca^{2+}$ release blockers (dantrolene and TMB-8) completely blocked these actions of lovastatin. In addition, the lovastatin-induced apoptosis was significantly reduced by a calpain inhibitor, a broad spectrum caspase inhibitor z-VAD-fmk and inhibitors specific for caspase-9 and caspase-3 (z-LEHD-fmk and z-DEVD-fmk, respectively), but not by an inhibitor specific for caspase-8 (z-IETD-fmk). Collectively, these results suggest that lovastatin induced apoptosis of HepG2 hepatoma cells through intracellular $Ca^{2+}$ release and calpain activation, leading to triggering mitochondrial apoptotic pathway. These results further suggest that lovastatin may be valuable for the therapeutic management of human hepatoma.

Diversity of Calpain-Calpastatin gene frequencies in Brown, Brindle and Jeju Black Hanwoo (한우, 칡소 및 제주 흑우 Calpain-Calpastatin 유전자 다양성)

  • Lee, Seung-Hwan;Kim, Seung-Chang;Cho, Soo-Hyun;Choi, Bong-Hwan;Sharma, A.;Lim, Dajeong;Dang, Chang-Gwan;Chang, Sun-Sik;Kim, Jae-Hwan;Ko, Moon Suk;Yang, Boh-Suck;Kang, Hee-Sul
    • Korean Journal of Agricultural Science
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    • v.40 no.2
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    • pp.147-153
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    • 2013
  • The aim of study was to investigate genetic diversity for the calpain/calpastatin gene in three Hanwoo breeds [(Brown (n=62), Brindle (n=81) and Jeju Black (n=30)]. Random samples from three breeds of Hanwoo were selected and genotyped for the 7 SNPs of calpain/calpastatin using TaqMan method. Allele frequencies were investigated for CAPN1/CAST gene. Allele frequency of CAST2 SNP was 0.75, 0.59 and 0.22 for Brown, Brindle and Jeju black, respectively. The CAST3 revealed allele frequency of 0.59 and 0.57 in Brown and Jeju Black, while it showed very low allele frequency (0.07) in Brindle. In particular, favorable allele (G allele) for the CAPN1-2 SNP which was shown a strong association with tenderness in Taurine and Indicine cattle revealed 16% and 17% higher allele frequency in Brown Hanwoo (0.82) comparing Brindle (0.66) and Jeju Black Hanwoo (0.65). AMOVA demonstrated that among population variance occupied only 10% of total variance and among individual variance was 0%, while within individual variance was 90% of total variance. This result showed that population effect contributed very small portion of genetic to these three Hanwoo breeds, while within individual variance contributed large portion of genetic diversity within these Hanwoo breeds. In conclusion, three Hanwoo breeds (Brown, Brindle and Jeju black) showed a genetically homogeneous based on the 7 SNPs of CAPN1/CAST gene and it came from same ancestor to form modern Hanwoo breed.