• Asian Pacific Journal of Cancer Prevention
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• v.18 no.11
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• pp.2919-2923
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• 2017

Objective: Anthocyanins belong to a class of flavonoids, exhibiting antioxidant and anti-inflammatory actions have been reported to have anti-cancer effects. Here, we investigated whether anthocyanins can inhibit cancer cell proliferation, invasion, and angiogenesis in human lung cancer A549 cells, which are critically involved in cancer metastasis. Methods: We used anthocyanins from fruits of Vitis coignetiae Pulliat (AIMs) which has been used in Korean folk medicine for the treatment of inflammatory diseases and cancers. We have performed cell proliferation assays, cell invasion assay, gelatin zymography, wound healing assay and western blotting to examine whether anthocyanins can inhibit cancer cell proliferation, invasion, and angiogenesis in A549 cells. Result: AIMs did not inhibit cancer cell proliferation on A549 cells. Also, AIMs suppressed cancer migration, and invasion by supressing MMP-2 and MMP-9 expression. The Immuno-blotting results also revealed that AIMs suppressed the proteins involved in cancer proliferation (COX-2, C-myc, cyclin D1), migration and invasion (MMP-2, MMP-9), anti-apoptosis (XIAP, and c-IAP2), adhesion and angiogenesis (ICAM-1, VEGF). Conclusion: This study demonstrates that the anthocyanins isolated from fruits of Vitis coignetiae Pulliat inhibit cancer proliferation, cancer migration, and invasion that is involve in cancer-metastasis. This study provides evidence that AIMs might have anti-cancer effects on human lung cancer.

• Journal of Veterinary Clinics
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• v.36 no.4
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• pp.200-206
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• 2019

This study was performed to assess the anti-inflammatory and cartilage regenerative effects of platelet-rich plasma (PRP) on canine chondrocytes. Proliferation and migration assays under both normal and lipopolysaccharide (LPS)-induced inflammatory conditions were performed with various concentrations of PRP (1% to 10%). The expression levels of genes related to osteoarthritis were evaluated in the following groups: PRP group, LPS group and LPS + PRP group. mRNA expression levels were detected using real-time polymerase chain reaction (RT-PCR). Proliferation assays showed significantly enhanced proliferation in all PRP-treated groups compared with the no serum group. Compared with 10% fetal bovine serum (FBS), PRP concentrations above 3% in the normal condition and 1% to 7% PRP in the LPS-induced inflammatory condition were found to significantly promote chondrocyte proliferation. In the normal condition, all PRP-treated groups showed significantly increased cell migration compared with the no serum group. Chondrocyte migration was decreased with LPS-induced inflammation, but PRP treatment resulted in significantly enhanced migration compared with the other groups in this condition. According to RT-PCR, the LPS + PRP group showed significantly higher levels of COL1A1, IL-6, aggrecan and lower levels of $TNF-{\alpha}$, MMP-1, MMP-3 mRNA expression compared to the LPS group. The results of this study suggest that PRP application can enhance the proliferation and migration of canine chondrocytes and improve canine articular cartilage regeneration.

• BMB Reports
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• v.55 no.5
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• pp.244-249
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• 2022

Characterized by abnormal proliferation and migration of vascular smooth muscle cells (VSMCs), neointima hyperplasia is a hallmark of vascular restenosis after percutaneous vascular interventions. Vaccinia-related kinase 1 (VRK1) is a stress adaption-associated ser/thr protein kinase that can induce the proliferation of various types of cells. However, the role of VRK1 in the proliferation and migration of VSMCs and neointima hyperplasia after vascular injury remains unknown. We observed increased expression of VRK1 in VSMCs subjected to platelet-derived growth factor (PDGF)-BB by western blotting. Silencing VRK1 by shVrk1 reduced the number of Ki-67-positive VSMCs and attenuated the migration of VSMCs. Mechanistically, we found that relative expression levels of β-catenin and effectors of mTOR complex 1 (mTORC1) such as phospho (p)-mammalian target of rapamycin (mTOR), p-S6, and p-4EBP1 were decreased after silencing VRK1. Restoration of β-catenin expression by SKL2001 and re-activation of mTORC1 by Tuberous sclerosis 1 siRNA (siTsc1) both abolished shVrk1-mediated inhibitory effect on VSMC proliferation and migration. siTsc1 also rescued the reduced expression of β-catenin caused by VRK1 inhibition. Furthermore, mTORC1 re-activation failed to recover the attenuated proliferation and migration of VSMC resulting from shVrk1 after silencing β-catenin. We also found that the vascular expression of VRK1 was increased after injury. VRK1 inactivation in vivo inhibited vascular injury-induced neointima hyperplasia in a β-catenin-dependent manner. These results demonstrate that inhibition of VRK1 can suppress the proliferation and migration of VSMC and neointima hyperplasia after vascular injury via mTORC1/β-catenin pathway.

• Journal of Ginseng Research
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• v.47 no.1
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• pp.133-143
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• 2023

Background: Past studies suggested that ginseng extracts and ginseng-derived molecules exerted significant regulatory effects on skin. However, no reports have described the effects of ginseng-derived nanoparticles (GDNPs) on skin cell proliferation and wound healing. In this study, we investigated whether GDNPs regulate the proliferation of skin cells and promote wound healing in a mouse model. Methods: GDNPs were separated and purified via differential centrifugation and sucrose/D2O gradient ultracentrifugation. GDNP uptake, cell proliferation and cell cycle progression were measured by confocal microscopy, CCK-8 assay and flow cytometry, respectively. Cell migration and angiogenic effects were assessed by the wound scratch assay and tube formation assay, respectively. ELISA was used to detect extracellular matrix secretion. The relevant signaling pathway was confirmed by western blotting. The effects of GDNPs on skin wound healing were assessed by wound observation, HE staining, and western blotting. Results: GDNPs possessed the essential features of exosomes, and they were accumulated by skin cells. Treatment with GDNPs notably enhanced the proliferation of HaCaT, BJ and HUVECs. GDNPs also enhanced the migration in HaCaT cells and HUVECs and angiogenesis in HUVECs. GDNPs increased the secretion of MMP-1, fibronectin-1, elastin-1, and COL1A1 in all three cell lines. GDNPs regulated cell proliferation through the ERK and AKT/ mTOR pathways. Furthermore, GDNPs facilitated skin wound healing and decreased inflammation in a mouse skin wound model. Conclusion: GDNPs can promote skin wound healing through the ERK and AKT/mTOR pathways. GDNPs thus represent an alternative treatment for chronic skin wounds.

• Animal Bioscience
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• v.36 no.11
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• pp.1757-1768
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• 2023

Objective: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. Methods: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17β-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. Results: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17β-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. Conclusion: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

• Food Science of Animal Resources
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• v.43 no.6
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• pp.1017-1030
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• 2023

Fetal bovine serum (FBS), which contains various nutrients, comprises 20% of the growth medium for cell-cultivated meat. However, ethical, cost, and scientific issues, necesitates identification of alternatives. In this study, we investigated commercially manufactured serum-free media capable of culturing Hanwoo satellite cells (HWSCs) to identify constituent proliferation enhancing factors. Six different serum-free media were selected, and the HWSC proliferation rates in these serum-free media were compared with that of control medium supplemented with 20% FBS. Among the six media, cell proliferation rates were higher only in StemFlex^TM Medium (SF) and Mesenchymal Stem Cell Growth Medium DXF (MS) than in the control medium. SF and MS contain high fibroblast growth factor 2 (FGF2) concentrations, and we found upregulated FGF2 protein expression in cells cultured in SF or MS. Activation of the fibroblast growth factor receptor 1 (FGFR1)-mediated signaling pathway and stimulation of muscle satellite cell proliferation-related factors were confirmed by the presence of related biomarkers (FGFR1, FRS2, Raf1, ERK, p38, Pax7, and MyoD) as indicated by quantitative polymerase chain reaction, western blotting, and immunocytochemistry. Moreover, PD173074, an FGFR1 inhibitor suppressed cell proliferation in SF and MS and downregulated related biomarkers (FGFR1, FRS2, Raf1, and ERK). The promotion of cell proliferation in SF and MS was therefore attributed to FGF2, which indicates that FGFR1 activation in muscle satellite cells may be a target for improving the efficiency of cell-cultivated meat production.

• The Journal of Internal Korean Medicine
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• v.18 no.1
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• pp.231-241
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• 1997

The purpose of this research was to investigate effect of water extract of Euphorbiae Pekinensis Radix and Moutan Cortex Radicis on the proliferation of human cancer cell-lines. The effects of Euphorbiae Pekinensis Radix and Moutan Cortex Radicis on the proliferation of A431, HeLa, MOLT-4, K562 cells, Balb/c 3T3 cells, mouse thymocytes, splenocytes and human lymphocytes were estimated by MTT colorimetric assay. The results were as follows; 1. In proliferation of A431, HeLa, MOLT-4 and K562 cell-lines, Euphorbiae Pekinensis Radix and Moutan Cortex Radicis inhibited the proliferation of K562 cells. 2. In the combined effect of Euphorbiae Pekinensis Radix and mitomycin C, Moutan Cortex Radicis and mitomycin C, all herbs stimulated the proliferation of MOL T-4 cells. 3. Euphorbiae Pekinensis Radix and Moutan Cortex Radicis did not inhibited the proliferation of Balb/c 3T3 cells. 4. Euphorbiae Pekinensis Radix and Moutan Cortex Radicis stimulated the proliferation of mouse thymocytes. 5. Euphorbiae Pekinensis Radix and Moutan Cortex Radicis stimulated the proliferation of mouse splenocytes. 6. Euphorbiae Pekinensis Radix and Moutan Cortex Radicis stimulated the proliferation of human lymphocytes.

• Journal of Animal Science and Technology
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• v.55 no.1
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• pp.19-23
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• 2013

The current study was carried out to determine the effects of Kohlrabi (Brassica oleracea var. gongylodes) on proliferation and differentiation of pig preadipocytes and $_3T_3-L_1$ cells. Pig preadipocytes were isolated from the backfat of the new-born pigs. Twenty-four hours after seeding, the cells were washed with DMEM/F-12 (designated day 0). To measure the cell proliferation, the cells were treated with 25 ng/ml and 100 ng/ml ethanol extracts of Kohlrabi (peel and flesh) for two days (day 0 ~ 2). To measure differentiation, the cells were treated with Kohlrabi for two days (day 0 ~ 2) and cell differentiation was measured on day 6. Twenty-five ng/ml and 100 ng/ml of Kohlrabi peel decreased proliferation of pig preadipocytes by 4.59% and 17.7%, respectively, compared with the control and Kohlrabi flesh by 11.4% and 19.2%, respectively. However, Kohlrabi did not inhibit cell differentiation. To measure the effects of Kohlrabi on proliferation and differentiation of $_3T_3-L_1$ cells, the cells were treated with Kohlrabi for two days in culture, like pig preadipocytes. Kohlrabi (both peel and flesh) did not show any effects on cell proliferation and differentiation. In summary, the results of the current study showed that Kohlrabi decreased proliferation of pig preadipocytes, but no inhibitory effects on differentiation of the cells. Kohlrabi had no effects on proliferation and differentiation of $_3T_3-L_1$ cells.

• Archives of Pharmacal Research
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• v.28 no.8
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• pp.948-955
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• 2005

Glomerular mesangial cells (GMCs) in diverse renal diseases undergo cell proliferation and/or hypertrophy, and gangliosides have been reported to play an important role in modulating cell structure and function. This study compared the effects of transforming growth $factor-\beta\; (TGF­\beta1)$ and the effects of the application of exogenous gangliosides on GMCs and investigated whether the application of exogenous gangliosides regulated cellular proliferation and hypertrophy. Human GMCs were cultured with exogenous gangliosides and $TGF-\beta1$ in a media containing $10\%$ fetal bovine serum and in a media without the fetal bovine serum. Exogenous gangliosides biphasically changed the proliferation of human GMCs (0.1-1.0 mg/mL). A low concentration (0.1 mg/mL) of gangliosides mainly increased the number of human GMCs, whereas cellular proliferation was significantly reduced by raising the concentration of exogenous gangliosides. $TGF-\beta1$ greatly reduced the number of human GMCs in a concentration­dependent manner (1-10 ng/mL). Serum deprivation accelerated the gangliosides- and $TGF­\beta1-induced$ inhibition of mesangial cell proliferation to a greater extent. Gangliosides (1.0 mg/ mL) and $TGF-\beta1$ (10 ng/mL) both caused a significant increase in the incorporation of $[^3H]leucine$ per cell in the serum-deprived condition, whereas it was completely reversed in serum­supplemented condition. Similar results to the $[^3H]leucine$ incorporation were also observed in the changes in cell size measured by flow cytometric analysis. These results show that exogenous gangliosides modulate cell proliferation and hypertrophy in cultured human GMCs, and these cellular responses were regulated differently based on whether the media contained serum or not. Results from the present study raise new possibilities about the potential involvement of gangliosides in the development of mesangial cell proliferation and hypertrophy.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.373-379
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• 2013

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or $37^{\circ}C$. In case of ICR strain, primary TSCs cultured at $37^{\circ}C$ showed significantly higher proliferation and viability than those at $35^{\circ}C$ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the $35^{\circ}C$ culture condition. Moreover, sub-passage of primary TSCs at $35^{\circ}C$ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at $35^{\circ}C$, which showed no significant difference in the viability, compared to those at $37^{\circ}C$. Furthermore, sub-passaged TSCs cultured at $37^{\circ}C$ showed no significant differences in proliferation and viability, compared to those at $35^{\circ}C$. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at $35^{\circ}C$, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at $37^{\circ}C$. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.