In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"The differential expression of lipocalin-2 in pig tissues and its molecular mechanism of regulating cell viability, differentiation, adipogenesis, and glycometabolism in porcine intramuscular preadipocytes.","authors":"Xiaoying Dong, Yanfei Chen, Liubing Tang, Qinen Wu, Tingjun Li, Qiuxia Wei, Shengqiu Tang","doi":"10.1007/s11626-026-01158-y","DOIUrl":"10.1007/s11626-026-01158-y","url":null,"abstract":"<p><p>Lipocalin-2 (LCN-2) is an adipocytokine secreted mainly by adipose tissue. Increasing evidences suggest that LCN-2 is an inflammatory factor associated with insulin resistance, obesity, and its complications. The precise mechanism of the development of obesity-related disorders induced by LCN-2 is not very clear. This study evaluated the expression of LCN-2 in pig tissues and its molecular mechanism of regulating preadipocyte differentiation in porcine intramuscular preadipocytes. LCN-2 expression in tissues of Tongcheng pigs, intramuscular adipose tissue of Tongcheng pigs (fat type) and Landrace pigs (lean type) in embryonic stage and growth stage, and adipocyte differentiation-induced porcine intramuscular preadipocytes was detected using reverse transcription polymerase chain reaction (qRT-PCR). After LCN-2 treatment, cell viability was measured by the methyl thiazolyl tetrazolium (MTT) method, mRNA expression of CCAAT/enhancer binding protein-ɑ (C/EBPɑ), adipocyte determination and differentiation factor-1 (ADD1), fatty acid desaturase (FAD), fatty acid synthase (FAS), and glucose transporter (GLUT) 1,4 was determined by qRT-PCR, protein expression of LCN-2 and peroxisome proliferator-activated receptor-γ (PPARγ) was analyzed by Western blot. Results of qRT-PCR indicated that LCN-2 showed significantly higher expression in high-intramuscular fat (IMF) pigs compared to low-IMF pigs (P < 0.05). LCN-2 expression in porcine intramuscular preadipocytes was significantly upregulated after adipocytic differentiation induction (P < 0.05). Silencing of LCN-2 with LCN-2 siRNA (siLCN-2) inhibited cell viability, lipid droplets, protein expression of PPARγ, and mRNA expression of C/EBPa, ADD1, FAD, FAS, and GLUT 1,4. siLCN-2 treated cells also showed a lower content of triglyceride and release of glucose. Moreover, LCN-2-induced downregulation of cell viability, adipocytic differentiation, adipogenesis, and glycometabolism of porcine intramuscular preadipocytes was partially blocked by the PPARγ inhibitor (GW9662). It is indicated that LCN-2 silencing suppresses cell viability, adipocytic differentiation and adipogenesis, glycometabolism, and fat deposition of porcine intramuscular preadipocytes through suppression of the PPARγ signaling pathway.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"525-536"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147473582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction of tissue-engineered vascular sinoatrial node in vitro.","authors":"Xiaolong Yu, Qing Chang, Lina Wang","doi":"10.1007/s11626-026-01159-x","DOIUrl":"10.1007/s11626-026-01159-x","url":null,"abstract":"<p><p>To develop a biological pacemaker by differentiating rabbit BMSCs into vascular endothelial cells (VECs) and pacemaker-like cells, and constructing a 3D vascularized sinoatrial node (SAN) in vitro. BMSCs were isolated via whole bone marrow adherence. Sinoatrial node cells were purified using differential adhesion with 5-BrdU treatment, and their lysate was prepared by freeze-thaw cycling. BMSCs were induced into VECs (CD31+/CD34+) using endothelial medium and into pacemaker-like cells (HCN2+/cTnT+) using SAN lysate. These cells were co-cultured on Matrigel at 1:1, 1:2, and 2:1 ratios to form 3D vascularized constructs. Cell distribution was analyzed via frozen sectioning and H&E staining. BMSCs successfully differentiated into VECs and pacemaker-like cells, confirmed by marker expression. The 1:1 co-culture ratio optimally promoted uniform cell distribution, network formation, and angiogenesis in the Matrigel scaffold. This study demonstrates that BMSCs can be differentiated into functional pacemaker-like cells and VECs, enabling the in vitro construction of a vascularized tissue-engineered SAN-a promising step toward biological pacemaker development.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"537-545"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms of Xinwei Tang in stress-induced gastric dysmotility: evidence from rat and In Vitro models.","authors":"Man Tian, Honghui Xiao, Yingbing Mei, Xinyu Zhang, Zhengliang Qi","doi":"10.1007/s11626-026-01151-5","DOIUrl":"10.1007/s11626-026-01151-5","url":null,"abstract":"<p><p>Stress is a key trigger of gastric dysmotility, partly via mitochondrial dysfunction and disordered gut-brain hormonal signaling. Xinwei Tang (XWT) is a multi-herb formula used empirically for upper gastrointestinal symptoms, but its mechanisms remain unclear. This study aimed to determine whether XWT alleviates water-immersion restraint stress (WIRS)-induced gastric dysmotility and to delineate underlying mitochondrial and metabolic pathways using integrated in vivo, in vitro and multi-omics approaches. Male rats underwent 7-d WIRS and received vehicle, domperidone (3 mg/kg) or XWT (3, 6, 12 g/kg). Gastric emptying, serum motilin/gastrin, oxidative stress indices and PINK1/Parkin-LC3/p62 proteins were assessed, and H₂O₂-injured GES-1 cells were treated with XWT-medicated serum. Gastric antra from MOD and XWT-H rats were analyzed by RNA-seq and DIA proteomics (n = 3/group). WIRS reduced gastric emptying by roughly half and lowered motilin/gastrin, increased ROS/MDA and disrupted PINK1/Parkin-LC3/p62 profiles; XWT dose-dependently reversed these changes, with XWT-H approximating domperidone. Omics revealed XWT-associated downregulation of inflammatory/protease and acute-phase genes/proteins and enrichment of oxidative phosphorylation, tricarboxylic-acid cycle and other metabolic pathways, without global activation of canonical autophagy/mitophagy gene sets. These preclinical data indicate that XWT ameliorates stress-induced gastric dysmotility via mitochondria- and metabolism-centred protection with selective tuning of mitophagy-related proteins.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"500-510"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147480590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oncogenic role and potential mechanisms of MYO1B in breast cancer.","authors":"Mei Dai, Qin Jiang, Danhua Zhang, Lun Li","doi":"10.1007/s11626-025-01149-5","DOIUrl":"10.1007/s11626-025-01149-5","url":null,"abstract":"<p><p>Myosin IB (MYO1B), a member of the type I myosin family, is overexpressed in various tumor tissues. MYO1B facilitates tumor progression by regulating cellular proliferation, migration, and the epithelial-mesenchymal transition (EMT). However, the precise function of MYO1B in breast cancer (BRCA) is still not well understood. We analyzed MYO1B expression, prognostic significance, immune infiltration, and its correlation with drug resistance in BRCA using meta-analysis and public databases. Functional assays in BRCA cells were performed to evaluate the effects of MYO1B on cell proliferation, apoptosis, and sensitivity to tamoxifen and palbociclib. MYO1B mRNA levels showed no significant difference between BRCA and normal tissues, whereas MYO1B protein was upregulated in tumor tissues. High MYO1B expression was associated with poor prognosis in BRCA patients. Functionally, MYO1B promoted BRCA cell proliferation, inhibited apoptosis, and increased resistance to tamoxifen and palbociclib. Mechanistically, MYO1B activated the Pi3k-AKT signaling pathway. Our study suggests that MYO1B promotes the progression of BRCA and may serve as a new target for overcoming endocrine therapy resistance.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"428-443"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147689611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A preliminary study on the mechanism of stromal interaction molecule 1 (STIM1) involvement in Adriamycin-induced podocyte injury.","authors":"Li Miao, Mi Bai, Songming Huang, Aihua Zhang, Siguang Lu","doi":"10.1007/s11626-026-01156-0","DOIUrl":"10.1007/s11626-026-01156-0","url":null,"abstract":"<p><p>Cellular metabolic reprogramming is intimately linked to various physiological and pathological processes. For instance, calcium (Ca²⁺)-mediated signaling pathways are essential for maintaining the homeostasis of critical cellular organelles. Stromal interaction molecule 1 (STIM1)-mediated store-operated calcium entry (SOCE) is a primary pathway for Ca²⁺ influx in non-excitable cells. This study aims to elucidate the role of STIM1 in podocyte injury. An STIM1 eukaryotic overexpression plasmid (p-STIM1) and small interfering RNA (si-STIM1) were constructed and separately transfected into mouse podocytes (MPC5). Flow cytometry was used to assess apoptotic rates, Fluo-3/AM calcium imaging to measure intracellular Ca²⁺ levels, and Western blotting to analyze the expression of endoplasmic reticulum stress (ERS)-related proteins. Additionally, mitochondrial morphology, membrane potential (MMP), reactive oxygen species (ROS) levels, and mitochondrial DNA (mtDNA) copy numbers were evaluated. Compared to STIM1 deficiency, STIM1 overexpression led to a marked increase in the apoptotic rate of Adriamycin-induced injured podocytes in vitro. This was associated with a significant rise in intracellular Ca²⁺ concentration and upregulation of ERS-related proteins, including GRP78, GRP94, and CHOP. Mitochondria displayed pronounced swelling and vacuole-like changes, a notable reduction in MMP, elevated ROS levels, and a decrease in mtDNA copies. STIM1 exacerbates podocyte injury by promoting intracellular Ca²⁺ influx, intensifying ERS, and inducing significant morphological and functional mitochondrial alterations. These findings suggest that targeting STIM1-mediated pathways could be a potential therapeutic strategy for podocyte-related kidney diseases.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"559-570"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147490929","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-495-3p attenuates cerebral ischemia-reperfusion-induced neuronal inflammation and apoptosis by targeting CCL2 expression.","authors":"XiaoDong Yu, LiZhi Xue, WenQin Zou, YanQing Deng, WenXin Jiang, GenShan Gao","doi":"10.1007/s11626-025-01148-6","DOIUrl":"10.1007/s11626-025-01148-6","url":null,"abstract":"<p><p>This study aimed to explore the function of miR-495-3p in cerebral ischemia-reperfusion injury (CI/RI) and reveal its potential molecular mechanism. In vivo and in vitro models of CI/RI were established by MACO/R and OGD/R, respectively. Neural function scores, HE staining, and TUNEL staining assessed the degree of brain tissue injury in mice. LDH assay, MTT assay, and flow cytometry evaluated neuronal toxicity, viability, and apoptosis rate. ELISA and Western blot evaluated inflammatory factors and the NF-κB pathway. Dual-luciferase reporting assay and RIP explored the targeting relationship between miR-495-3p and CCL2. miR-495-3p was abnormally low in MACO/R mouse brain tissue and OGD/R-damaged neurons, while CCL2 was highly expressed. miR-495-3p overexpression improved neuronal apoptosis and inflammation in the brain tissue of MACO/R mice. Consistent results were also obtained in in vitro experiments. Enhancing CCL2 or knocking down miR-495-3p aggravated OGD/R-induced neuronal damage. The deleterious effects of miR-495-3p knockdown were prevented by the knockdown of CCL2. miR-495-3p targeted CCL2. miR-495-3p improves CI/R-mediated neuronal apoptosis and inflammation through targeted regulation of CCL2 expression. These results provide data support for CI/RI-targeting drugs and the understanding of disease mechanisms.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"489-499"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DDX3 inhibits PTZ-induced ferroptosis in human neuronal SH-SY5Y cells via Wnt/β-catenin signaling.","authors":"Mian Zou, Yanhui Zhou, Congcong Zhang, Guoshuai Yang","doi":"10.1007/s11626-025-01138-8","DOIUrl":"10.1007/s11626-025-01138-8","url":null,"abstract":"<p><p>Ferroptosis is a novel form of programmed cell death characterized by the accumulation of lipid peroxides and associated with neuropathic diseases. However, the molecular mechanisms remain unclear. This study aimed to investigate the potential mechanism of DDX3 in pentylenetetrazole (PTZ)-induced ferroptosis in human neuronal SH-SY5Y cells. PTZ induced SH-SY5Y cells to simulate the neuropathic disease model in vitro. Western blot analysis was used to assess DDX3, β-catenin, β-catenin phosphorylated at Ser37/Thr41, GPX4, and ACSL4 expression. Nuclear accumulation of β-catenin was tested by IF. MMP-7, c-Myc, cyclin D1, LEF1, and Axin2 were detected by qRT-PCR. Cell viability was measured by CCK-8. Apoptosis was detected by flow cytometry. Total antioxidant status (TAS) and total oxidant status (TOS) levels were detected by biochemical kit. ROS production was detected by flow cytometry. Biochemical kits were used to detect MDA, 4-HNE, Fe²⁺, and GSH levels. Our results showed that DDX3 expression was decreased in PTZ-induced SH-SY5Y cells. DDX3 overexpression promoted PTZ-induced SH-SY5Y cell viability, inhibited apoptosis, promoted TAS and GSH expression, and inhibited TOS, MDA, 4-HNE, Fe²⁺, and ROS levels, indicating that DDX3 reduced PTZ-induced SH-SY5Y cell ferroptosis. DDX3 knockdown reduced total β-catenin protein, nuclear accumulation of β-catenin, Wnt target genes (MMP-7, c-Myc, cyclin D1, LEF1, and Axin2), and GPX4 expression in PTZ-induced SH-SY5Y cells, while increasing β-catenin phosphorylated at Ser37/Thr41 and ACSL4 expression. The effect of DDX3 overexpression on the above indexes was opposite to that of DDX3 knockdown. β-catenin overexpression and Wnt/β-catenin signaling activator CHIR99021 increased total β-catenin protein, nuclear accumulation of β-catenin, MMP-7, c-Myc, cyclin D1, LEF1, and Axin2 expression in PTZ-induced SH-SY5Y cells, while decreasing β-catenin phosphorylated at Ser37/Thr41expression and ACSL4 expression. In addition, β-catenin overexpression and CHIR99021 increased cell viability, reduced apoptosis, and upregulated TAS, GSH, and GPX4 expression, while decreasing TOS, MDA, 4-HNE, Fe²⁺, ROS, and ACSL4 levels. GPX4 knockdown and ACSL4 overexpression reversed β-catenin overexpression effects. Further results showed that DDX3 inhibited PTZ-induced SH-SY5Y cell ferroptosis by activating Wnt/β-catenin signaling. Our results suggested that DDX3 inhibited PTZ-induced ferroptosis in SH-SY5Y cells through activation of Wnt/β-catenin signaling. Our findings may provide new molecular targets for the treatment of neuropathic diseases.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"459-475"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147456946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcription factor HOXA4 promotes HBV replication and hepatocellular carcinoma proliferation by activating KIF11 transcription.","authors":"Jinling Ma, Duodi Liao","doi":"10.1007/s11626-025-01144-w","DOIUrl":"10.1007/s11626-025-01144-w","url":null,"abstract":"<p><p>KIF11 is a mitotic kinesin responsible for the formation and maintenance of bipolar spindles, and it has high expression in the hepatocellular carcinoma (HCC). However, the role of the KIF11 gene in the hepatitis B virus (HBV)-related HCC remains unknown. Thus, this study aims to explore the function of transcription factor HOXA4 binding to KIF11 in HBV-related HCC, with the goal of providing a novel gene therapy approach for its treatment. HBV-positive (HepG2.2.15 cells) and HBV-negative (HepG2 cells) liver cancer cells were used to investigate the expression of KIF11 and HOXA4. HepG2 cells or HepG2.2.15 cells were transfected with the pc3.1-HBx, si-KIF11, and si-HOXA4, or co-transfected with the si-HOXA4 and oe-KIF11 for subsequent analysis. The binding of HOXA4 protein to the KIF11 gene promoter was examined based on a ChIP assay. The characteristics of HepG2.2.15 cells and HepG2 cells were assessed using CCK-8 and flow cytometry. HBV transcription and replication levels were detected via Northern and Southern blotting. The secretion level of HBV antigens in the HepG2.2.15 cell supernatant was measured by ELISA. KIF11 and HOXA4 were highly expressed in the HepG2.2.15 cells. Silencing KIF11 inhibited cell viability and HBV replication and transcription, reduced HBsAg and HBeAg levels in cell supernatants, promoted apoptosis, and downregulated p-PI3K/PI3K and p-AKT/AKT protein expression in HepG2.2.15 cells. pc3.1-HBx promoted cell viability, inhibited apoptosis, and upregulated p-PI3K/PI3K and p-AKT/AKT protein expression in HepG2 cells, which was reversed by si-KIF11. ChIP assays confirmed that HOXA4 bound to the KIF11 gene promoter. Silencing HOXA4 suppressed cell viability and HBV replication and transcription, decreased HBsAg and HBeAg levels in cell supernatants, enhanced apoptosis, and downregulated p-PI3K/PI3K and p-AKT/AKT protein expression in HepG2.2.15 cells. Silencing HOXA4 inhibited cell viability, promoted apoptosis, and downregulated p-PI3K/PI3K and p-AKT/AKT protein expression in HepG2 cells with pc3.1-HBx transfection. Overexpression of KIF11 counteracted the effects of si-HOXA4 in the HepG2.2.15 cells or HepG2 cells with pc3.1-HBx transfection. In conclusion, silencing of HOXA4 inhibited HBV replication and HCC proliferation by downregulating KIF11, providing a novel gene-assisted therapeutic approach for HBV-related HCC.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"546-558"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146179500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The abnormal thyroxine signals triggers podocyte apoptosis in DN mice.","authors":"Jing Wang, Binfang Huang, Guangquan Zhou","doi":"10.1007/s11626-026-01153-3","DOIUrl":"10.1007/s11626-026-01153-3","url":null,"abstract":"<p><p>Podocyte injury is a pivotal factor in the advancement of diabetic nephropathy (DN). The present study aimed to delineate the influence of disrupted thyroxine signaling on podocyte apoptosis in DN mouse models. We employed bioinformatics analyses, coupled with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment assessments, to identify differentially expressed genes (DEGs) associated with thyroxine signaling in both human and murine DN datasets. Subsequently, we elucidated the function of thyroid hormone receptor α1 (THRA1) and nuclear receptor co-repressor 1 (NCOR1) on glomerular injury and podocytes apoptosis under hypothyroid and hyperglycemic conditions, respectively. Our findings highlight that hypothyroidism significantly alters glomerular gene expression profiles in DN mice leading to increased podocyte apoptosis. This effect occurs through a dual mechanism: on one hand, the upregulation of THRA1 expression induced by DN results in direct glomerular injury, which was further aggravated by hypothyroidism; on the other hand, the downregulation of NCOR1 expression thereby increases THRA1 activity levels. Our data suggests that disturbed thyroxin signals could trigger podocyte apoptosis and glomerular injury in DN mice, offering new insights into DN pathogenesis while laying groundwork for innovative therapeutic strategies.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"444-458"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of ARL4C antisense oligonucleotide with reduced off-target effects and enhanced efficacy as an anti-cancer drug.","authors":"Kanae Kawai, Shinji Matsumoto, Akikazu Harada, Ryota Sada, Takeshi Harada, Yuuya Kasahara, Satoshi Obika, Yasushi Okamura, Akira Kikuchi","doi":"10.1007/s11626-026-01154-2","DOIUrl":"10.1007/s11626-026-01154-2","url":null,"abstract":"<p><p>ARL4C, a member of the small GTP-binding protein superfamily, is overexpressed in the tumor lesions of various cancers. Antisense oligonucleotide (ASO) therapy targeting ARL4C (ASO-1316-A), a 15-mer ASO, has been shown to inhibit xenograft tumor formation. To enhance therapeutic efficacy and reduce off-target effects, fourteen 18-mer ARL4C-targeting ASOs were developed. Among them, ASO-2025-A/L demonstrated superior suppression of ARL4C mRNA and protein expression compared to ASO-1316-A. In vitro, ASO-2025-A/L inhibited the proliferation, migration, and adhesion abilities of PANC-1 and S2-CP8 pancreatic cancer cells as well as PC-9 lung cancer cells more effectively than ASO-1316-A. ASO-2025-A/L was predicted to reduce the expression of 26 potential off-target genes, compared to 2824 potential off-target genes affected by ASO-1316-A in PANC-1 cells. While ASO-2025-A/L downregulated 317 genes, which were confirmed by RNA-sequence analysis, the majority was involved in cell adhesion pathways downstream of ARL4C signaling, and only two genes corresponded to off-target genes. In vivo, intravenous administration of ASO-2025-A/L inhibited xenograft tumor growth induced by PC-9 and PANC-1 cells, accompanied by a reduction in ARL4C mRNA levels in tumors without elevating serum liver toxicity markers. These findings highlight the development of an improved 18-mer ARL4C-targeting ASO with enhanced therapeutic efficacy and reduced off-target effects.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"413-427"},"PeriodicalIF":2.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147369162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}