STAR Protocols
·
2026
·Vol. 7
·期 4
·104854
·
IF 1.4
Yan Fang,Jianfeng Shen
Steps for preparing LIN28B siRNA-loaded nanoparticles • Procedures for establishing a murine malignant ascites model • Instructions for administering nanoparticles and PARP inhibitors in vivo • Methods for assessing tumor burden and vascular permeability Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Summary Malignant serous effusion (MSE) is a life-threatening complication of advanced cancers with limited effective treatments. Here, we present a protocol for the preparation of LIN28B siRNA-loaded nanoparticles and their combination with PARP inhibition for therapeutic evaluation in malignant ascites (MA). We describe steps for nanoparticle formulation, establishment of murine ascites models, and in vivo administration of therapeutic agents. We then detail procedures for assessing tumor progression, vascular permeability, and endothelial junction integrity to evaluate treatment efficacy and microenvironmental remodeling. For complete details on the use and execution of this protocol, please refer to Fang et al. 1 Download: Download high-res image (221KB) Download: Download full-size image
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高影响因子
ACS Applied Nano Materials
·
2026
·
IF 5.8
Liu Kai,Shen Hongliang,Li Jietao,Feng Yu,Zhou Chang,Jiang Anlong
Osteosarcoma remains a highly aggressive primary bone
malignancy,
and its clinical treatment is frequently limited by chemoresistance,
systemic toxicity, and insufficient tumor-specific drug accumulation.
To address these challenges, we developed an OS-7.9 aptamer-functionalized
ZIF-8 nanoscale platform for the targeted codelivery of sorafenib
(SFN) and GPX4 siRNA, designated O-SR-ZIF-8, with the intended application
of ferroptosis-based osteosarcoma therapy. The resulting nanoparticles
exhibited a uniform nanoscale size distribution, favorable colloidal
stability, high payload-loading capacity, and pH-responsive release
behavior. The ZIF-8 nanostructure remained relatively stable under
physiological conditions but promoted accelerated release of SFN and
GPX4 siRNA in acidic tumor-mimicking microenvironments, supporting
its suitability for tumor-localized delivery. Compared with nontargeted
formulations, O-SR-ZIF-8 markedly enhanced cellular internalization
in MG-63 osteosarcoma cells, inhibited cell proliferation, promoted
cell death, increased intracellular oxidative stress, and suppressed
migration and invasion. In a murine osteosarcoma xenograft model,
systemic administration of O-SR-ZIF-8 significantly restrained tumor
growth through GPX4 silencing and modulation of ferroptosis-related
signaling. Moreover, the nanoscale formulation exhibited favorable
biocompatibility, as evidenced by negligible hemolytic activity and
no obvious histopathological injury in major organs. Together, these
findings indicate that O-SR-ZIF-8 is an aptamer-guided nanoscale therapeutic
platform with potential in the treatment of osteosarcoma through targeted
delivery of sorafenib and GPX4 siRNA to silence genes and increase
oxidative stress associated with ferroptosis.
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Small
·
2025
·
IF 12.1
Shuangjun Li,Shengnan Li,Wen Li,Yue Hua,Ruyue Shi,Dongmei Zhao,Jiaxin Wang,Xiuhua Zhao
Oral delivery of nucleic acid therapeutics for cancer therapy encounters major challenges, such as gastrointestinal acidity, enzymatic degradation, mucus barriers, and P-glycoprotein (P-gp) efflux. To address these challenges, this study engineers virus-like nanoparticles encapsulating siCENPN nucleic acid complexes and irradiated tumor cells (ITC), further coated with zwitterionic bacterial cellulose derivatives (PB-BC-BY). This innovative formulation demonstrates enhanced mucus penetration, improves stability in gastrointestinal environments, and effective inhibition of P-gp-mediated efflux. The nanoparticles effectively deliver siRNA into tumor cells, resulting in marked downregulation of CENPN expression and strong autophagy induction. The autophagic tumor cells and ITCs subsequently stimulated peripheral blood mononuclear cells (PBMCs) to secrete anti-tumor cytokines, thereby triggering an immune response. In vivo experiments confirm that orally administered nanoparticles successfully traverse the gastrointestinal barrier, enter systemic circulation, target tumors, and significantly inhibit tumor growth. Enhanced activation of dendritic cells, along with increased infiltration of cytotoxic T lymphocytes (CTLs) and Th1 cells in the tumor microenvironment, further confirms effective immune activation. Moreover, this treatment induces strong immune memory, prolonges survival, and provides substantial protection against tumor recurrence. This study presents a novel oral delivery strategy for nucleic acid therapeutics, offering an effective approach by simultaneously inducing tumor cell autophagy and promoting a robust immune-mediated anti-cancer response.
研究领域:
纳米医学
·
RNA治疗学
·
癌症生物学
·
药物递送
·
免疫治疗
·
生物化学
·
生物物理学
·
分子生物学
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PubMed
高影响因子
Biomaterials Science
·
2021
·Vol. 9
·期 15
·5275-5292
·
IF 6.843
Xinyue Zhang,Kai Hong,Qingmei Sun,Yunqing Zhu,Jianzhong Du
RNA interference (RNAi) technology has great potential in cancer therapy, e.g., small interfering RNA (siRNA) can be exploited to silence specific oncogenes related to tumor growth and progression. However, it is critical to achieve high transfection efficiency while reducing cytotoxicity. In this paper, we report an siRNA delivery strategy targeting the oncogene KRAS based on arginine-modified poly(disulfide amine)/siRNA nanocomplexes. The poly(disulfide amine) is synthesized via aza-Michael polyaddition followed by the introduction of arginine groups onto its backbone to afford poly((N,N′-bis(acryloyl)cystamine-co-ethylenediamine)-g-Nω-p-tosyl-L-arginine) (PBR) polycations. Thus multiple interactions including electrostatic interaction, hydrogen bonding and a hydrophobic effect are introduced simultaneously between PBR and siRNA or cell membranes to improve transfection efficiency. By optimizing the grafting density of arginine groups, PBR/siRNA nanocomplexes achieve high cellular uptake efficiency, successful endosomal/lysosomal escape, and rapid biodegradation in the presence of high GSH concentration in the cytoplasm, and finally release siRNA to activate the RNAi mechanism. Additionally, compared to commercially available PEI 25K, PBR/siRNA nanocomplexes possess a significantly increased gene silencing effect on human pancreatic cancer cells (PANC-1) with decreased cytotoxicity and enhanced tumor penetration ability in PANC-1 multicellular spheroids in vitro. Overall, with both GSH-responsiveness and excellent tumor penetration, this safe and efficient poly(disulfide amine)-based siRNA delivery system is expected to provide a new strategy for gene therapy of pancreatic cancer and other stromal-rich tumors.
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高影响因子
Journal of Materials Chemistry B
·
2015
·Vol. 4
·期 3
·547-556
·
IF 4.726
Mengyi Li,Xiaoyan Zhou,Xiaolong Zeng,Changyong Wang,Jiake Xu,Dong Ma,Wei Xue
For effective gene delivery to breast cancer MCF-7 cells, a folate-targeting redox gene carrier was synthesized by Michael addition polymerization between 1-(2-aminoethyl)piperazine and N,N′-cystaminebisacrylamide. Folate was then conjugated through an amidation reaction. The obtained folate-modified hyperbranched poly(amido amine)s (FA-PAAs) degraded in the presence of glutathione and displayed excellent transfection efficiency in vitro. In particular, FA-PAAs showed much higher gene delivery efficiency than PEI-25k in the presence of serum, leading to an obvious decrease in MMP-9 protein expression and the apoptosis of MCF-7 cells. Moreover, FA-PAAs displayed lower cytotoxicity and better blood compatibility than PEI-25k, suggesting a potential application in gene therapy for tumors.
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高影响因子
INTERNATIONAL JOURNAL OF PHARMACEUTICS
·
2023
·
IF 5.8
Chang Liu,Xiaojun Zhang,Huiying Yang,Meijun Zhao,Yanhong Liu,Risheng Zhao,Ziqing Li,Meng Sun
Ischemic heart disease (IHD) is a cardiac disorder in which myocardial damage occurs as a result of myocardial ischemia and hypoxia. Evidence suggests that oxidative stress and inflammatory responses are critical in the development of myocardial ischemia. Therefore, the combination of antioxidant and anti-inflammatory applications is an effective strategy to combat ischemic heart disease. In this paper, polyethylene glycol (PEG)-modified cationic liposomes were used as carriers to deliver apigenin (Apn) with small interfering RNA (siRNA) targeting the receptor for glycosylation end products (RAGE) (siRAGE) into cardiomyocytes to prevent myocardial ischemic injury through antioxidant and anti-inflammatory effects. Our results showed that we successfully prepared cationic PEG liposomes loaded with Apn and siRAGE (P-CLP-A/R) with normal appearance and morphology, particle size and Zeta potential, and good encapsulation rate, drug loading and in vitro release degree. In vitro , P-CLP-A/R was able to prevent oxidative stress injury in H9C2 cells, downregulate the expression of RAGE, reduce the secretion of cellular inflammatory factors and inhibit apoptosis through the RAGE/NF-κB pathway; In vivo , P-CLP-A/R was able to prevent arrhythmia and myocardial pathological injury, and reduce apoptosis and the area of necrotic myocardium in rats. In conclusion, P-CLP-A/R has a protective effect on myocardial ischemic injury and is expected to be a potential drug for the prevention of ischemic heart disease in the future.
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高影响因子
DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY
·
2020
·
IF 2.365
Zhongjuan Wang,Yueqin Liang,Yanqiu Liu,Hongying Xia,Jianqi Liu,Xingfang Jin,Zhongkun Li
Multi-drug resistance (MDR) is one of the major challenges in the successful chemotherapy of non-small cell lung cancer (NSCLC). Although RNA interference (RNAi) has been widely used to silence resistance-related genes, the effect remains unsatisfactory. In this study, we attempted to overcome MDR of NSCLC by simultaneously interfering with two RNAs that have different functions. A new pH-triggered polyglutamate brush polymer dimethylmaleic anhydride-poly(ethyleneglycol) monomethyl ether-b-polyglutamate-g-spermine (DMA-mPEG-b-PG-g-spermine, DPPGS) was designed and synthesized. The DPPGS/small interfering RNA (siRNA) complex nanoparticles (DPPGSN) were prepared. The results demonstrated that DPPGSN could be transformed from a negatively charged form into a positively charged form in the slightly acidic tumor extracellular environment. The siRNA targeting MDR1 mRNA (siMDR1) and siRNA targeting survivin mRNA (siSurvivin) could be efficiently co-delivered by DPPGS to simultaneously interfere with two genes (p < 0.01). Furthermore, DPPGS co-delivery of siMDR1 and siSurvivin lowered the IC50 value of cisplatin (DDP) in A549/DDP (p < 0.01) cells and increased the apoptosis rate of the cells (p < 0.01). Therefore, co-delivery of siMDR1 and siSurvivin using DPPGS would be a promising approach for overcoming MDR of NSCLC.
研究领域:
肿瘤学
·
药剂学
·
分子生物学
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PubMed
高影响因子
PLoS One
·
2017
·Vol. 12
·期 8
·e0183941
·
IF 2.806
Wen Che,Yun Bao,Feng Tang
Background
Resistance to human epidermal growth factor receptor-2 (HER2)-targeted therapies is common, and results in treatment failure and new tumor progression. C35 is over-expressed in many invasive breast cancer endogenously, and functions as an oncogene in breast cancer cell lines.
Methods
The current study aims to investigate effects of C35 on cell viability and migration of HER2-positive breast tumor cells and explore possible mechanisms. The protein expression of C35 in BT-474 breast ductal carcinoma cells was interfered by siRNAs. MTT assay was utilized to detect the viability of BT474 breast ductal carcinoma cells after transfection. The migration of BT474 breast ductal carcinoma cells was examined by cell wound scratch assay.
Results
In addition, the protein expression of C35, HER2, AKT and p-AKT in breast ductal carcinoma cells were detected by Western Blot. The protein expression of C35 was weaker in siRNA1 and siRNA2 groups compared with the control group. The relative expression ratio of C35 to β-actin decreased significantly in siRNA2 and siRNA1 groups. When C35 expression was interfered by siRNA1 and siRNA2 for 48 h, the viability of BT474 breast ductal carcinoma cells decreased compared with the control group. In addition, the migration of breast ductal carcinoma cells decreased when C35 expression was interfered by siRNA1 for 24 h, and interfered by siRNA1 and siRNA2 for 48 h.
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
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高影响因子
Small Methods
·
2020
·Vol. 5
·期 4
·2001047
·
IF 12.13
Jianqiao Chang,Yu Zhang,Yike Li,Ziwei Han,Fei Tian,Chao Liu,Qiang Feng,Yuguang Wang,Jiashu Sun,Lu Zhang
Detection of mRNA expression in live cells during treatment is a challenging task, despite its importance in tumor biology and potential therapeutic leads. Here a multilayer ratiometric fluorescent nanomachine for live-cell perturbation and imaging of mRNA at single cell resolution is reported. The nanomachines fabricated by microfluidic approaches consist of fluorescent polymeric cores and multiple lipid layers, which can efficiently deliver siRNA and molecular beacons (MBs) to cytosol and then release the cargo in a sequential way. The siRNA molecules released from the outer lipid layers lead to silencing of multidrug resistance 1 (MDR1) gene, and the MBs from the middle lipid layers detect the presence of MDR1 mRNA. The fluorescent ratio of MBs to fluorescent polymeric cores positively correlates with the expression level of MDR1 mRNA in MCF-7/ADR cells during siRNA treatment. The nanomachines provide comparable results with traditional qPCR for quantifying mRNA, showing great potential for modulation and imaging of intratumoral mRNA in vitro and in vivo.
研究领域:
生物技术
·
分子生物学
·
癌症研究
·
纳米技术
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高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Enyang Yao,Yang Li,Kai Deng,Wei He,Milad Ashrafizadeh,João Conde,Liyu Yang
Osteosarcoma is the most common primary bone tumor with a high metastatic rate. Despite the improvement in patient prognosis with conventional therapeutics, enhancing the survival of patients with metastatic disease remains a challenge. Zoledronic acid (ZOL) stimulates apoptosis in cancer cells and is considered a promising drug for osteosarcoma therapy. Moreover, depletion of Inhibin Subunit Beta A (INHBA), a member of the transforming growth factor superfamily, can suppress osteosarcoma cell proliferation and invasion. Regarding the restricted potential of monotherapy, combination therapies, such as small interfering RNA (siRNA) and chemotherapy, are ideal strategies. However, off-target effects and adverse impacts lead to unsatisfactory clinical efficacy of these drugs. To address this challenge, we designed a folic acid-modified macrophage membrane coated with a carbon quantum dot (CQD) nanoplatform for the co-delivery of ZOL and siINHBA (ZOL-siINHBA@CQD@RM-FA). The anticancer activity of this nanotherapeutic agent was examined in vitro and in vivo . ZOL-siINHBA@CQD@RM-FA was approximately 200.7 nm in size, had a negative charge, excellent pH responsiveness, desirable biocompatibility, and high cellular uptake. This nanosystem achieved favorable co-delivery of ZOL/siINHBA and impaired osteosarcoma proliferation and invasion in vitro . Notably, intravenous administration facilitated tumor growth regression and inhibited lung metastasis without systemic toxicity. Accordingly, this nanotherapy could be considered an effective treatment for osteosarcoma, and the nanodelivery system shows potential for combined therapy by co-delivering genes and chemotherapy drugs.
研究领域:
肿瘤学
·
纳米医学
·
药物递送系统
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高影响因子
EXPERIMENTAL ANIMALS
·
2023
·
IF 2.4
Jianqiang WEI,Jing YIN,Ying CUI,Kaijie WANG,Mingyan HONG,Jianzhong CUI
Intracerebral hemorrhage (ICH) is an incurable neurological disease. Microglia activation and its related inflammation contribute to ICH-associated brain damage. FERM domain containing kindlin 1 (FERMT1) is an integrin-binding protein that participates in microglia-associated inflammation, but its role in ICH is unclear. An ICH model was constructed by injecting 50 μl of autologous blood into the bregma of rats. FERMT1 siRNA was injected into the right ventricle of the rat for knockdown of FERMT1. A significant striatal hematoma was observed in ICH rats. FERMT1 knockdown reduced the water content of brain tissue, alleviated brain hematoma and improved behavioral function in ICH rats. FERMT1 knockdown reduced microglia activity, inhibited NLR family pyrin domain containing 3 (NLRP3) inflammasome activity and decreased the expression of inflammatory factors including interleukin-1β (IL-1β) and interleukin 18 (IL-18) in the peri-hematoma tissues. BV2 microglial cells were transfected with FERMT1 siRNA and incubated with 60 μM Hemin for 24 h. Activation of NLRP3 inflammasome induced by hemin were reduced in microglia when FERMT1 was knocked down, leading to decreased production of inflammatory factors IL-1β and IL-18. In addition, knockdown of FERMT1 prevented the activation of nuclear factor kappa B (NF-κB) signaling pathway in vivo and in vitro. Our findings suggested that down-regulation of FERMT1 attenuated microglial inflammation and brain damage induced by ICH via NLRP3/NF-κB pathway. FERMT1 is a key regulator of inflammatory damage in rats after ICH.
研究领域:
神经科学
·
分子生物学
·
炎症
·
神经病学
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高影响因子
Journal of the American Chemical Society
·
2026
·
IF 16.6
Xiaofei Xin,Yifu Lyu,Huanyu Qin,Jinyu Dai,Di Wu,Yu Ding,Qiang Tian,Chao Qin,Na Li,Lifang Yin
The rational design of nonlamellar, multicompartment lipid nanoparticles (LNPs) offers a promising strategy for advancing nucleic acid delivery and immunotherapy. In this study, imidazole-based LNPs (A3-DM/DL-LNPs) with a mesoscopically disordered internal structure resembling the sponge phase were developed, enabling efficient codelivery of mRNA and STAT3-targeting siRNA (siSTAT3). Small-angle scattering analysis revealed a unique quasi-periodic arrangement in A3-DM/DL-LNPs, characterized by broader interdomain spacing and higher membrane heterogeneity compared to benchmark ALC-0315-LNPs, suggesting enhanced endocytosis efficiency and more favorable RNA encapsulation. The “M-shaped” polar headgroup, formed by a piperazine ring flanked by two imidazole moieties, promoted membrane interaction, cellular uptake, and transfection efficiency. In contrast, alternative LNPs with linear polar head structures (A3-SS/SM/SL-LNPs and A4-SS/SM/SL-LNPs) showed inferior delivery performance. Raman spectroscopy revealed that lipid spatial localization correlated with RNA expression in the spleen and lymph nodes, highlighting the importance of LNP structure in immune activation and targeting specific immune organs. Functionally, A3-DM/DL-LNPs restored dendritic cell (DC) antigen presentation, alleviated endoplasmic reticulum (ER) stress, and reversed T cell exhaustion in acute myeloid leukemia (AML), triggering strong immune responses and enhancing natural killer (NK) cell and T cell-mediated antileukemic activity, thereby improving therapeutic outcomes.
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高影响因子
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