STAR Protocols
·
2024
·
IF 1.3
Xiaoxiang Gao,Yinghui Qiu,Wei Liao,Chao Zhao
Summary Regulating cancer-related microRNAs (miRNAs) may become a new generation of therapeutic modalities for cancer treatment. Here, we describe a protocol based on hepatoma-22 (H22) tumor-bearing mice to screen potential targets for treating hepatocellular carcinoma (HCC). We detail the construction of H22 tumor-bearing mice and treatment with two natural compounds, Ulva lactuca L. polysaccharide (ULP) and 5-fluorouracil (5FU). We further describe the isolation of the tumor tissues for miRNA sequencing and the discovery and validation of potential miRNA gene targets against HCC. For complete details on the use and execution of this protocol, please refer to Qiu et al. 1
研究领域:
肿瘤学
·
分子生物学
·
遗传学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Orthopaedic Surgery and Research
·
2023
·Vol. 18
·期 1
·1-15
·
IF 2.6
Lin Yuan,Dai Hanhao,Yu Guoyu,Song Chao,Liu Jun,Xu Jie
ObjectiveOsteoporosis is a progressive systemic skeletal disorder. Multiple profiling studies have contributed to characterizing biomarkers and therapeutic targets for osteoporosis. However, due to the limitation of the platform of miRNA sequencing, only a part of miRNA can be sequenced based on one platform.Materials and methodsIn this study, we performed miRNA sequencing in osteoporosis bone samples based on a novel platform Illumina Hiseq 2500. Bioinformatics analysis was performed to construct osteoporosis-related competing endogenous RNA (ceRNA) networks. Gene interference and osteogenic induction were used to examine the effect of identified ceRNA networks on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (HBMSCs).Results miR-1303 was lowly expressed, while cochlin (COCH) and KCNMA1-AS1 were highly expressed in the osteoporosis subjects. COCH knockdown improved the osteogenic differentiation of HBMSCs. Meanwhile, COCH inhibition compensated for the suppression of osteogenic differentiation of HBMSCs by miR-1303 knockdown. Further, KCNMA1-AS1 knockdown promoted osteogenic differentiation of HBMSCs through downregulating COCH by sponging miR-1303.Conclusion sOur findings suggest that the KCNMA1-AS1/miR-1303/COCH axis is a promising biomarker and therapeutic target for osteoporosis.
研究领域:
分子生物学
·
基因组学
·
干细胞研究
·
骨骼生物学
·
治疗靶点
✓ 已验证
DOI
PubMed
高影响因子
LAB ON A CHIP
·
2021
·Vol. 21
·期 9
·1759-1770
·
IF 6.799
Wenwen Chen,Rongkai Cao,Wentao Su,Xu Zhang,Yuhai Xu,Peng Wang,Zhongqiao Gan,Yingying Xie,Hongjing Li,Jianhua Qin
Tumor-derived exosomes have been recognized as promising biomarkers for early-stage cancer diagnosis, tumor prognosis monitoring and individual medical treatment. However, it is a huge challenge to separate exosomes from trace biological samples in clinics for disease diagnosis. Herein, we propose a simple, quick, and label-free method for isolating circulating exosomes from serum of patients. The strategy synergistically integrates chitosan electrostatic-adsorption, micro-patterned substrates, and microfluidic shuttle flow control to enable the capture/release of circulating exosomes in a simple manner. Using this microchip, we can isolate exosomes from trace samples (10 μl) with relative purity over 90% and high RNA recovery ratio over 84% within 15 minutes, which is impossible for traditional ultracentrifugation methods. We then validate the application of the microchip using 24 serum samples from clinical breast cancer and breast fibroma patients. The isolated exosomes are subjected to miRNA sequencing and RT-PCR, followed by pathway prediction analysis. The results showed that exosomes were relevant to the invasion and metastasis of breast cancer cells and hsa-miR-18a-3p might have the potential to become a new biomarker for distinguishing breast cancer from breast fibroma (AUC = 0.83, P value = 0.019). This established method is simple, quick and easy to operate with integration. And it may pave a new way for clinical research on exosomes and tumor relevant diagnosis.
研究领域:
生物医学工程
·
癌症生物学
·
分子生物学
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Neural Regeneration Research
·
2023
·Vol. 18
·期 10
·2291
·
IF 6.1
Zhang Zhong-Xia,Zhou Yong-Jie,Gu Ping,Zhao Wei,Chen Hong-Xu,Wu Ruo-Yu,Zhou Lu-Yang,Cui Qing-Zhuo,Sun Shao-Kang,Zhang Lin-Qi,Zhang Ke,Xu Hong-Jun,Chai Xi-Qing,An Sheng-Jun
Microglia -mediated inflammatory responses have been shown to play a crucial role in Parkinson’s disease . In addition, exosomes derived from mesenchymal stem cells have shown anti-inflammatory effects in the treatment of a variety of diseases. However, whether they can protect neurons in Parkinson’s disease by inhibiting microglia -mediated inflammatory responses is not yet known. In this study, exosomes were isolated from human umbilical cord mesenchymal stem cells and injected into a 6-hydroxydopamine -induced rat model of Parkinson’s disease . We found that the exosomes injected through the tail vein and lateral ventricle were absorbed by dopaminergic neurons and microglia on the affected side of the brain, where they repaired nigral-striatal dopamine system damage and inhibited microglial activation. Furthermore, in an in vitro cell model, pretreating lipopolysaccharide-stimulated BV2 cells with exosomes reduced interleukin-1β and interleukin-18 secretion, prevented the adoption of pyroptosis -associated morphology by BV2 cells, and increased the survival rate of SH-SY5Y cells. Potential targets for treatment with human umbilical cord mesenchymal stem cells and exosomes were further identified by high-throughput microRNA sequencing and protein spectrum sequencing. Our findings suggest that human umbilical cord mesenchymal stem cells and exosomes are a potential treatment for Parkinson’s disease , and that their neuroprotective effects may be mediated by inhibition of excessive microglial proliferation.
研究领域:
神经科学
·
干细胞生物学
·
炎症
·
分子生物学
·
神经退行性疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Aquaculture Reports
·
2023
·
IF 3.7
Hui Li,Shuangfeng Li,Liming Zhao,Shuying Zhang,Jing Zhang,Xianfeng Wang,Junping Liang
Cottonseed meal has been one of the predominant protein feedstuffs used in aqua feed, which contains gossypol . It is well known that gossypol, a natural polyphenolic compound found in seeds, can be toxic to fish. The toxicity of gossypol persists even after a recovery period; however, the underlying mechanism remains largely unknown. To investigate the toxic mechanism of gossypol after the recovery period, Cyprinus carpio were administered a continuous oral dose of 20 mg/kg·d gossypol (gossypol acetate dose/fish weight·d) for one week, followed by a two-week recovery period. The spleen index, spleen morphology, lysozyme (LZM) activity, acid phosphatase (ACP) activity and nitric oxide (NO) concentration were obviously changed in the spleens of C. carpio after two weeks of recovery. Then, miRNA sequencing identified 22 differentially expressed miRNAs in the spleens of C. carpio two weeks after the gossypol treatment. The mRNA expression levels of immune-associated genes were significantly increased in the spleens after recovery. MiR-194 was chosen for future study through a search of existing literature and an analysis of miRNA and mRNA expression levels. To further characterize the potential target of miR-194, we identified putative miR-194 binding sites in the 3′UTR of myeloid differentiation factor 88a (MyD88a). Dual-luciferase reporter assays revealed that miR-194 downregulated the expression level of MyD88a through binding to seed-matching sequence in the 3′ UTR of MyD88a. Moreover, C. carpio were intraperitoneally administered with miR-194 antagomir. Furthermore, down-regulation of miR-194 increased the expression levels of MyD88a and some pro-inflammatory cytokines and anti-inflammatory cytokines. Altogether, the results demonstrate miR-194 can target MyD88a to regulate the immune response of C. carpio after a two-week recovery period following gossypol administration.
研究领域:
水产养殖
·
毒理学
·
免疫学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）