• Animal Bioscience
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• 제37권3호
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• pp.509-521
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• 2024

Objective: It was shown that microRNAs (miRNAs) play an important role in milk protein synthesis. However, the post-transcriptional regulation of casein expression by exogenous miRNA (xeno-miRNAs) in ruminants remains unclear. This study explores the regulatory roles of alfalfa xeno-miR162 on casein synthesis in bovine mammary epithelial cells (bMECs). Methods: The effects of alfalfa xenomiR-162 and G protein subunit gamma 11 (GNG11) on proliferation and milk protein metabolism of bMECs were detected by 5-Ethynyl-2'-Deoxyuridine (EdU) staining, flow cytometry, cell counting kit-8 (CCK-8), enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Dual-luciferase reporter assay was used to verify the targeting relationship between GNG11 and xenomiR-162. Results: Results showed that over-expression of xenomiR-162 inhibited cell proliferation but promoted apoptosis, which also up-regulated the expression of several casein coding genes, including CSN1S1, CSN1S2, and CSN3, while decreasing the expression of CSN2. Furthermore, the targeting relationship between GNG11 and xenomiR-162 was determined, and it was confirmed that GNG11 silencing also inhibited cell proliferation but promoted apoptosis and reduced the expression of casein coding genes and genes related to the mammalian target of rapamycin (mTOR) pathway. Conclusion: Alfalfa xenomiR-162 appears to regulate bMECs proliferation and milk protein synthesis via GNG11 in the mTOR pathway, suggesting that this xeno-miRNA could be harnessed to modulate CSN3 expression in dairy cows, and increase κ-casein contents in milk.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2001년도 전기 제11차 학술대회 논문집
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• pp.35-37
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• 2001

인간 Lactadherin은 비유 개시와 동시에 mammary gland의 epithelial cell에 의해 생성되는 46 Kd의 glycoprotein이다. 이 단백질은 성장기 어린이의 diarrheal disease의 원인이 되는 rotavirus에 specific하게 결합하여 그 증식을 억제시켜서 바이러스성 설사를 예방한다. 본 연구에서는 한국 여성의 lactadherin gene을 cloning 하여 그 sequence를 분석 비교하고 서양 여성과의 유전자 변이를 조사하였다. 보고된 서양 여성의 lactadherin 유전자와 비교하여 한 여성의 유전자에는 silent mutation을 포함하여 3 곳의 mutation이 있었고, 또 다른 여성의 유전자에서 새로운 3 곳의 mutation이 존재하여 이 유전자의 SNP가 여러 곳에 산재되어 있음이 관찰되었다.

• Journal of Dairy Science and Biotechnology
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• 제15권1호
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• pp.27-32
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• 1997

Most lipids in milk are dispetsed as the form of fat globules. Apical portion of plasma membrane is coated with fat globules, which are synthesized from mammary epithelial cells and then secreted into the lumen. The unique phenomenon in separation of the plasma membrane from the cell is observed only in mammary system. It has been suggested that milk fat globule membrane(MFGM) is formed from endoplasmic reticulum, Golgi apparatus, secretory granule to plasma membrane. For this reason MFGM is important for nuderstanding the structure and function of biological membrane. Because MFGM also plays an important role in inhibition of lipase action, stimulation of nutrient digestion and absorption, emulsion or function as natural liposome, study of the major components in MFGM will provide the opportunity for more broad industrial uses of MFGM in the future.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.59-59
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• 2001

Interleukin-10 (IL-10) is known as a regulator of inflammation and pathogenesis in mammalian organs, but its precise role is little known in the mammary gland. Our initial experiment showed that IL-10 expression levels in mice decreased at the lactation stage otherwise increased at the involution stage. To reveal the effects of IL-10 on the involution of mammary gland, expression profiles of the apoptosis-related genes were examined in transgenic mice expressing human IL-10 as well as in knock-out mice (IL-10-/-). Mild inflammatory legions by lymphocytes were observed in the mammary glands of transgenic lines at the lactation stage. The expression of TRAIL (Tumor necrosis factor-Related Apoptosis-Inducing Ligand) among the apoptosis-related genes was highly elevated in the transgenic mice while others were not significantly changed. Furthermore, TRAIL was down regulated by four fold in the IL-10-/- mice at the involution stage. The expression of DR4 was elevated at the involution stage of normal mice. DR4 was detected in the milk of transgenic mice but absent in that of normal mice. Our results proposed that the elevated IL-10 at the involution stage recruit lymphocytes and induce TRAIL and DR4 genes, therefore, lead to enter involution stage of mammary glands.

• Molecules and Cells
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• 제39권5호
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• pp.410-417
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• 2016

During the lactation cycle of the bovine mammary gland, autophagy is induced in bovine mammary epithelial cells (BMECs) as a cellular homeostasis and survival mechanism. Interferon gamma ($IFN-{\gamma}$) is an important antiproliferative and apoptogenic factor that has been shown to induce autophagy in multiple cell lines in vitro. However, it remains unclear whether $IFN-{\gamma}$ can induce autophagy and whether autophagy affects milk synthesis in BMECs. To understand whether $IFN-{\gamma}$ affects milk synthesis, we isolated and purified primary BMECs and investigated the effect of $IFN-{\gamma}$ on milk synthesis in primary BMECs in vitro. The results showed that $IFN-{\gamma}$ significantly inhibits milk synthesis and that autophagy was clearly induced in primary BMECs in vitro within 24 h. Interestingly, autophagy was observed following $IFN-{\gamma}$ treatment, and the inhibition of autophagy can improve milk protein and milk fat synthesis. Conversely, upregulation of autophagy decreased milk synthesis. Furthermore, mechanistic analysis confirmed that $IFN-{\gamma}$ mediated autophagy by depleting arginine and inhibiting the general control nonderepressible-2 kinase (GCN2)/eukaryotic initiation factor $2{\alpha}$ ($eIF2{\alpha}$) signaling pathway in BMECs. Then, it was found that arginine supplementation could attenuate $IFN-{\gamma}$-induced autophagy and recover milk synthesis to some extent. These findings may not only provide a novel measure for preventing the $IFN-{\gamma}$-induced decrease in milk quality but also a useful therapeutic approach for $IFN-{\gamma}$-associated breast diseases in other animals and humans.

• 한국가시화정보학회지
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• 제13권1호
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• pp.43-48
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• 2015

Collective cell migration is a fundamental phenomenon observed in various biological processes such as development, wound healing, and cancer metastasis. During the collective migration, cells undergo changes in their phenotypes from those of stable to the migratory state via the process called epithelial-mesenchymal transition (EMT). Recent findings in biology and biochemistry have shown that EMT is closely related to the cancer invasion or metastasis, but not much of the correlations in kinematics and physical forces between the neighboring cells are known yet. In this study, we aim to understand the cell migration and stress distribution within the expanding cell cluster. We constructed the in vitro cell cluster on the hydrogel, employed traction force microscopy (TFM) and monolayer stress microscopy (MSM) to visualize the physical forces within the expanding cell monolayer. During the expansion, cells at the cluster edge exhibited enhanced motility and developed focal adhesions that are the essential features of EMT while cells at the core of the cluster maintained the epithelial characteristics. In the aspect of mechanical stress, the cluster edge had the highest traction force of ~90 Pa directed toward the cluster core, which means that cells at the edge actively pull the substrate to make the cluster expansion. The cluster core of the tightly confined cells by neighboring cells had a lower traction force value (~60 Pa) but the highest intercellular normal stress of ~800 Pa because of the accumulation of traction from the edge of the monolayer.

• 생명과학회지
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• 제31권2호
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• pp.248-255
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• 2021

유선종양은 암컷 개에서 가장 흔한 암 중의 하나이며 종양 타입에 따라 조직학적으로 다양한 종류의 세포들이 존재한다. 특히, 복합 상피암종(complex carcinoma)의 경우 내강상피세포(luminal epithelium)와 근상피세포(myoepithelium)가 혼재되어 종양 내 세포 이형성(intra-tumoral heterogeneity)을 보인다. 하지만, 이러한 다양한 종양세포의 기원과 종양의 악성화에 미치는 영향에 대해서는 아직 밝혀진 바 없다. 최근, 여러 종류의 사람 종양에서 알려진 종양줄기세포는 종양 내 세포의 다양성(diversity)에 관여하고 악성화에도 기여할 수 있다고 보고되었다. 흥미롭게도 종양줄기세포는 자가재생능과 다분화능을 갖는 정상줄기세포와 동일한 능력을 가지고 있지만 종양 특이 유전자의 돌연변이와 줄기세포성격유지를 조절하는 신호전달체계에 문제가 있어 종양의 발생 시작부터 다른 조직으로의 전이에 관여하여 개체의 생존률에 직접적인 영향을 준다. 뿐만 아니라, 방사선 및 화학항암제에 대한 저항성을 보이기 때문에 종양 재발에 밀접하게 연관되어 있다. 본 리뷰 논문은 개 유선종양의 특성 및 종류, 종양줄기세포의 정의, 분리 방법, 임상학적 중요성에 대해서 정리하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권4호
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• pp.576-584
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• 2002

Bovine PAS-4 is an integral membrane glycoprotein expressed in mammary epithelial cells. Complementary DNA (cDNA) cloning of PAS-4 was performed by reverse-transcriptase polymerase chain reaction (RT-PCR) with oligonucleotide probes based on it's amino terminal and internal tryptic-peptides. The cloned PAS-4 cDNA was 1,852 nucleotides (nt) long and its open reading frame (ORF) was encoded 1,413 base long. The deduced amino acid sequence indicated that PAS-4 consisted of 471 amino acid residues with molecular weight of 52,796, bearing 8 potential N-glycosylation sites and 9 cysteine residues. Partial bovine CD36 cDNA from liver also was sequenced and the homology of both nucleotide sequence was 94%. Most of the identical amino acid residues were in the luminal/extracellular domains. Contrary to PAS-4, bovine liver CD36 displays 6 potential N-glycosylation sites, which were located, except for those at positions 101 and 171, at same positions as PAS-4 cDNA. Cysteine residues of PAS-4 and CD36 were same at position and in numbers. Northern blot analysis showed that PAS-4 was widely expressed, although its mRNA steady-state levels vary considerably among the analyzed cell types. PAS-4 possessed hydrophobic amino acid segments near the amino- and carboxyl-termini. Two short cytoplasmic tails of the amino- and carboxyl-terminal ends constituted of a 5-7 and 8-11 amino acid residues, respectively.

• 한국임상수의학회지
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• 제26권6호
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• pp.556-562
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• 2009

피부와 유선 종양은 개에서 매우 빈번히 발생하는 종양으로, 종양 발생의 위험은 나이가 증가함에 따라 종양의 발생률은 증가하며, 발병률과 발병부위의 분포는 종양의 종류마다 다르다. 피부와 유선종양의 발생은 공기오염, 식생활 등 환경의 변화와도 관련이 있어 이들 종양들의 발생양상은 그 사회의 환경을 반영할 수 있어 시대와 지역에 따라 다양하게 다르게 나타날 수 있다. 따라서, 그 동안의 많은 보고들에도 불구하고, 피부와 유선종양발생에 대한 자료들은 종양에 대한 수의임상학적 진단과 치료를 위한 중요한 정보를 제공한다. 본 연구에서는 2005년 1월 1일부터 2006년 12월 31일까지 수의과학검역원과 강원대학교에 의뢰된 128증례의 개 피부종양과 240증례의 개 유선종양을 Goldschmidt와 Misdorp의 형태학적 기준에 의해 조직병리학적으로 진단하여, 종별, 성별 및 부위별로 각각 분류하여 국내에서 이들 종양의 발생양상을 체계적으로 분석하였다. 피부종양에서는 피부상피세포종양, 샘종양 및 간질종양이 각각 14.1%, 41.4%와 44.5%였다. Cutaneous histiocytoma가 피부종양에서 가장 빈번하게 관찰되어 전체 피부종양의 25.0%를 차지하였으며, 기름샘종 (sebaceous gland adenoma; 15.6%), 기저세포종(basal cell tumor; 8.6%), 항문둘레샘종 (perianal gland adenoma; 6.3%)와 지방종 (lipoma; 6.3%)순으로 높았다. 그 외에 비만세포종, 모낭세포종과 중층편평상피암 등이 관찰되었다. 이러한 피부종양들은 품종, 성과 나이 또는 부위에 따라 유사하거나 다른 양상을 나타내었다. 유선종양의 경우 양성과 악성이 201/240, 39/240으로 양성발생률이 현저히 높았으며, 양성혼합종 (benign mixed tumor)이 검사한 품종 모두에서 가장 높은 발생률 (35.0%)을 나타내었다. Complex adenoma는 Maltese와 Poodle에서 특히 높았으며, 악성종양에서는 simple carcinoma와 complex carcinoma가 각각 6.7%로, 악성 중 41.0%를 차지하여 가장 높은 발생률을 나타내었다. 본 연구결과는 최근 우리나라에서 여러 품종의 반려견에서 발생한 피부와 유선종양의 발생양상을 조직병리학적으로 진단 분석한 것으로, 임상수의사들이 지속적으로 증가추세에 있는 개의 피부와 유선종양을 진단하고 치료하는데 유용하게 활용될 수 있기를 기대한다.

• 대한세포병리학회지
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• 제18권1호
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• pp.69-73
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• 2007

Mammary carcinoma with osteoclast-like giant cells is an unusual neoplasm characterized by giant cells, mononuclear stromal cells, and hemorrhage accompanying a low grade carcinoma. We present the cytological findings in a case of invasive ductal carcinoma with osteoclast-like giant cells that was initially confused with a fibroadenoma, due to its well-demarcated and soft mass and the young age of the patient. A 28-year-old female presented with a 4.5 cm, well demarcated, soft and nontender mass in the right breast. Fine needle aspiration cytology (FNAC) showed a combination of low grade malignant epithelial cell clusters and osteoclast-like giant cells. The atypical epithelial cells were present in cohesive sheets and clusters. Osteoclast-like giant cells and bland-looking mononuclear cells were scattered. An histological examination revealed the presence of an invasive ductal carcinoma with osteoclast-like giant cells. We report here the cytological findings of this rare carcinoma in a very young woman. The minimal atypia of the epithelial cells and its soft consistency may lead to a false negative diagnosis in a young woman. The recognition that osteoclastlike giant cells are rarely present in a low grade carcinoma, but not in benign lesion, can assist the physician in making a correct diagnosis.