• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.46-46
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• 2003

Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1206-1212
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• 2023

The Wnt/β-catenin pathway plays essential roles in regulating various cellular behaviors, including proliferation, survival, and differentiation [1-3]. The intracellular β-catenin level, which is regulated by a proteasomal degradation pathway, is critical to Wnt/β-catenin pathway control [4]. Normally, casein kinase 1 (CK1) and glycogen synthase kinase-3β (GSK-3β), which form a complex with the scaffolding protein Axin and the tumor suppressor protein adenomatous polyposis coli (APC), phosphorylate β-catenin at Ser45, Thr41, Ser37, and Ser33 [5, 6]. Phosphorylated β-catenin is ubiquitinated by the β-transducin repeat-containing protein (β-TrCP), an F-box E3 ubiquitin ligase complex, and ubiquitinated β-catenin is degraded via a proteasome pathway [7, 8]. Colorectal cancer is a significant cause of cancer-related deaths worldwide. Abnormal up-regulation of the Wnt/β-catenin pathway is a major pathological event in intestinal epithelial cells during human colorectal cancer oncogenesis [9]. Genetic mutations in the APC gene are observed in familial adenomatous polyposis coli (FAP) and sporadic colorectal cancers [10]. In addition, mutations in the N-terminal phosphorylation motif of the β-catenin gene were found in patients with colorectal cancer [11]. These mutations cause β-catenin to accumulate in the nucleus, where it forms complexes with transcription factors of the T-cell factor/lymphocyte enhancer factor (TCF/LEF) family to stimulate the expression of β-catenin responsive genes, such as c-Myc and cyclin D1, which leads to colorectal tumorigenesis [12-14]. Therefore, downregulating β-catenin response transcription (CRT) is a potential strategy for preventing and treating colorectal cancer. Plant cytokinins are N⁶-substituted purine derivatives; they promote cell division in plants and regulate developmental pathways. Natural cytokinins are classified as isoprenoid (isopentenyladenine, zeatin, and dihydrozeatin), aromatic (benzyladenine, topolin, and methoxytopolin), or furfural (kinetin and kinetin riboside), depending on their structure [15, 16]. Kinetin riboside was identified in coconut water and is a naturally produced cytokinin that induces apoptosis and exhibits antiproliferative activity in several human cancer cell lines [17]. However, little attention has been paid to kinetin riboside's mode of action. In this study, we show that kinetin riboside exerts its cytotoxic activity against colon cancer cells by suppressing the Wnt/β-catenin pathway and promoting intracellular β-catenin degradation.

• Journal of Nutrition and Health
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• 제2권4호
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• pp.173-181
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• 1969

생후 $40{\pm}5$ 일된 젖떨어진 Albino Rat female male 각각 30마리를 크게 Alanine(AL)과 Aminoethylphosphonic acid(AEP) group으로 나누고 각 group을 다시 각각 2 group으로 나누어 female male Alanine 0.1%, female male Alanine 0.2%와 female male AEP 0.1%, female male AEP 0.2% 합(合) 8 group으로 나누어 14 주동안 실험한 결과의 요약은 다음과 같다. 단백질 효율과 뇨중 질소배설량에 어떤 유의적인 차이가 없음으로 AEP도 다른 일반적인 Amino acid와 같은 정도로 체내에서 이용됨을 알 수 있다. AEP의 첨가로 인한 p의 이용율의 변화 유무를 판정하기 위하여 뇨중 p의 함량을 측정한 결과 모든 group사이에 유의성이 없었음으로 alanine 첨가 group에서나 AEP 첨가 group에서나 같은 정도로 체내에서 이용됨을 알 수있으나 AEP의 첨가로 인한 Dietary Phosphorus함량이 높음으로 인하여 체내 보유량이 많다는 것을 알 수 있으며 이는 blood phosphorus 함량측정을 해본 결과와 일치함을 알 수 있다. 간질소 함량을 측정하여 본 결과 AEP 첨가가 간질소 함량을 높일 수 있으며 0.2%쪽이 더 많은 양을 간에 보유할 수 있다는 것을 알았다. 간지방 함량도 간질소 함량과 같은 방향으로 나타났으며 조직 검사로 fat lumps를 발견할 수 없었으므로 AEP 첨가가 fatty liver의 요인이 될 수 없다는 것을 알 수 있다. Amino acid의 생물학적 가치와 Amino acid의 조성사이에는 현저한 관계가 있어 양적으로 충분한 단백질을 섭취한다 하더라도 섭취한 단백질이 함유하고 있는 Amino acid의 종류 및 각종 Amino acid의 비율에 따라서 소화에 현저한 차이가 있어 생물학적 기능이 다르다는 것이 Oser와 그외 여러 학자들에 의하여 보고되었다. 이 보고에 비추어 보아 0.2% diet에서 Amino acid의 장내 흡수시 imbalanced condition을 초래 한 것으로 본다.