https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/564 2026-04-12T17:42:23Z https://scholarworks.korea.ac.kr/kumedicine/handle/2021.sw.kumedicine/77622 Title: Gene expression network analysis identified CDK1 and KIF11 as possible key molecules in the development of colorectal cancer from normal tissues Authors: Lee, Soo Bin; Noh, Young Seon; Moon, Ji-Wook; Sim, Soohyun; Han, Sung Won; Kim, Eun Sun; Lee, Ji-Yun Abstract: Background: Colorectal cancer (CRC) is one of the most common malignancies and the second most common cause of cancer-related mortality worldwide. Despite extensive research, the mechanism underlying CRC development remains unclear. This study aimed to understand the development and progression of CRC. Methods: Gene network analysis of tumors with their paired normal tissues was performed using the differentially expressed genes dataset for CRC from the Cancer Genome Atlas. Further investigation of the regulatory relationship between hub genes and tumor development was conducted by protein–protein interaction network, Gene Ontology enrichment, and Kyoto Encyclopedia of Genes and Genomes pathway analyses using the selected hub genes. Results: The network was more centered, and a common hub as well as a hub of hub genes were more connected to each other in the tumor than in the normal tissue, indicating changes in the network from normal to tumor. Eight downregulated and two upregulated hub genes (CDK1 and KIF11) in the tumor were identified. Further, the regulatory pathway was altered, especially in cell cycle and cell division. All R implementation codes are available on the journal website as supplementary materials. Conclusions: Our findings may help understand the biological processes underlying tumor development and progression and suggest CDK1 and KIF11 as possible key molecules in the development of CRC. © The Author(s) 2025. 2025-12-01T00:00:00Z https://scholarworks.korea.ac.kr/kumedicine/handle/2021.sw.kumedicine/80350 Title: Protocol for in vitro differentiation of C2C12 myoblasts into mature myotubes Authors: Oh, Jong Min; Lyu, Ji Hyo; Sihombing, Maic Audo Eybi Mayer; Bae, Chang-Hwan; Shin, Ung Cheol; Muthamil, Subramanian; Kim, Seon-Wook; Azim, M. N.; Kim, Jong-Hoon; Park, Jun Hong Abstract: BackgroundMouse C2C12 myoblast cell line is widely used as a good in vitro model to study muscle physiology, metabolism, and regeneration. The in vitro model is capable of replicating the main steps of skeletal muscle development, which may help in investigating the molecular mechanism behind myogenesis, muscle degeneration, especially in cancer patients, and in the atrophy of muscles in older people.ObjectiveConsidering the in vitro muscle differentiation study, there is a need to develop an ideal protocol for C2C12 myoblast differentiation.ResultsC2C12 myoblasts were differentiated into myotubes within 4 days when cultured in Dulbecco's modified Eagle's medium (DMEM) with 2% horse serum, which represents the key feature of myogenesis. Before differentiation, C2C12 cells were cultured up to 90-100% confluence, then the medium was replaced with differentiation media. Then, the cells started to differentiate, on day 2 (D2); the cells were fused with neighboring cells to form long cells. On day 3, the cells became more elongated, and on day 4, myotubes were clearly detected, having a cylindrical and elongated shape. The cells can be maintained at this stage for more than a week with regular media changes.ConclusionThis protocol could be helpful for translational research related to cancer, genetic disorders, and age-related muscle-wasting diseases. 2025-12-01T00:00:00Z https://scholarworks.korea.ac.kr/kumedicine/handle/2021.sw.kumedicine/78445 Title: Meso-macroporous hydrogel for direct litre-scale isolation of extracellular vesicles Authors: Kim, Junbeom; Kang, Minjin; Han, Geonhee; Hyung, Sujin; Kim, Mina; Jang, Minjeong; Lee, Han Kyul; Seo, Yunhee; Gil, Ki Cheol; Kim, Changheon; Song, Sojin; Jeong, Seonghyeok; Kim, Seongchan; Kim, Min Soo; Shim, Ji Sung; Kang, Sung Gu; Lee, Young Chan; Chung, Seok; Cho, Il-Joo; Shim, Tae Soup; Song, Kwang Hoon; Min, Jouha; Seong, Hyejeong; Lee, Kyungeun; Lee, Jeeyun; Lee, Cheolju; Kim, Hong Nam; Lee, Hyojin; Kim, Sun Hwa; Kang, Ji Yoon; Bong, Ki Wan; Choi, Nakwon Abstract: Extracellular vesicles are cell-originated lipid bilayer membrane vesicles that play vital roles in cell-to-cell communications. While extracellular vesicles hold substantial biomedical potential, conventional methodologies for isolating extracellular vesicles require elaborate preprocessing and, therefore, remain labour intensive and limited by throughput. To overcome these challenges, we present a facile fabrication route for generating a meso-macroporous hydrogel matrix with pores of similar to 400 nm for customizable extracellular vesicle isolation. By combining surface charge-selective capture of extracellular vesicles within the hydrogel matrix and their recovery by high ionic strength, we report direct extracellular vesicle isolation with a throughput range from microlitre to litre scales, without preprocessing, for various biofluids, including whole blood, plasma, ascites, saliva, urine, bovine milk and cell culture media. Furthermore, we demonstrate that the meso-macroporous hydrogel also serves as a solid-phase matrix for preserving extracellular vesicles for on-demand downstream analyses, making it applicable for therapeutics, cosmeceuticals and disease diagnostics. 2025-11-01T00:00:00Z https://scholarworks.korea.ac.kr/kumedicine/handle/2021.sw.kumedicine/78165 Title: Alpha-Ketoisocaproate Attenuates Muscle Atrophy in Cancer Cachexia Models Authors: Lim, Pooreum; Woo, Sang-woo; Han, Jihye; Lee, Young-lim; Lim, Jin-ju; Kang, Yeonghoon; Moon, Ji-wook; Nam, Jeong-min; Kim, Jeong-hyeon; Kim, Donghun; Shim, Jae Ho; Kim, Hyeon Soo Abstract: Background: Cancer-associated cachexia (CAC) is a multifactorial syndrome characterised by progressive loss of muscle mass with limited Food and Drug Administration treatments. Although emerging evidence suggests that l-leucine and β-hydroxy-β-methyl butyrate (HMB) have potential for treating CAC, the role of α-ketoisocaproate (KIC), a metabolite of l-leucine, remains unclear. Therefore, this study explored the use of KIC as a therapeutic agent for CAC-induced muscle atrophy by targeting myostatin. Methods: We evaluated the effect of KIC on muscle atrophy using BALB/c mice and C2C12 myotubes as models of C26- and 4T1-induced CAC. Male and female mice were injected with C26 and 4T1 cells, respectively. Grip strength was measured weekly, and mice were sacrificed 4 weeks post-injection for muscle collection. C2C12 myotubes were treated with conditioned media (CM) derived from C26 or 4T1 cells. Results: KIC suppressed mRNA expression of myostatin, a key regulator of muscle atrophy, more effectively than did l-leucine (−26.37 ± 4.11%, p < 0.01). KIC enhanced protein turnover in C2C12 myotubes and maintained 50% cell viability at high concentrations (KIC: 4.68 mM, HMB: 3.11 mM). Following CM treatment, KIC suppressed MuRF1 and MAFbx expression in a myostatin-dependent manner, thereby reducing their polyubiquitination. KIC restored Akt-FoxO3a phosphorylation, leading to improved myotube diameter (+63.8 ± 25.71%, p < 0.05) and fusion index (+51.9 ± 22.6%, p < 0.05). Immunofluorescence and nuclear fractionation revealed that KIC reduced FoxO3a nuclear accumulation. CM reduced p-Akt–FoxO3a interaction, which was rescued by KIC. In vivo, KIC administration increased body weight (11.11 ± 8.53%), grip strength (24.76 ± 10.58%), and skeletal muscle mass (p < 0.001) in C26 tumour-bearing mice. Protein expression of myostatin in the tibialis anterior (TA) muscle (−23.57 ± 12.22%, p < 0.05) and serum (−52.11 ± 3.56%, p < 0.001) was lower in KIC-treated mice (n = 12) compared with that in the controls. KIC increased the mean fibre cross-sectional area in TA (24.51 ± 14.14%, p < 0.01). In 4T1 tumour-bearing mice, KIC improved body weight (13.10 ± 10.76%) and grip strength (7.42 ± 4.33%) (p < 0.001, n = 10). Serum myostatin levels (−57.43 ± 9.46%, p < 0.001) and skeletal muscle weight were reduced in KIC-treated mice (n = 10). Conclusion: Our findings demonstrate that KIC improves muscle function in CAC-induced muscle atrophy by regulating myostatin expression in skeletal muscle via the Akt–FoxO3a pathway. Thus, KIC has been proposed as a potential therapeutic agent against CAC. © 2025 Elsevier B.V., All rights reserved. 2025-08-01T00:00:00Z