Selective removal of ovarian cancer cells from human ascites fluid using magnetic nanoparticles

A majority of ovarian cancer metastases result from the shedding of malignant cells from the primary tumor into the abdominal cavity. Free-floating cancer cells in serous effusions of late-stage ovarian cancer patients may spread to internal organs, making effective treatment extremely difficult. Selective removal of ovarian cancer cells from serous fluids may abate metastasis and improve long-term prognoses. We have already shown that superparamagnetic nanoparticles conjugated to an ephrin-A1 mimetic peptide with a high affinity for the EphA2 receptor can be used to capture and remove cultured human ovarian cancer cells from the peritonea of experimental mice. Here we demonstrate the potential clinical utility of the methodology by in vitro capture and isolation of cancer cells from the ascites fluid of ovarian cancer patients.From the Clinical EditorOvarian cancer metastases usually are the result of shedding of malignant cells from the primary tumor into the abdominal cavity. In this paper, a novel nanotechnology-based method is demonstrated for the in vitro capture and isolation of cancer cells from the ascites fluid of ovarian cancer patients.     

Molecular characterization of scant lung tumor cells using iron-oxide nanoparticles and micro-nuclear magnetic resonance

Advances in nanotechnology and microfluidics are enabling the analysis of small amounts of human cells. We tested whether recently developed micro-nuclear magnetic resonance (μNMR) technology could be leveraged for diagnosing pulmonary malignancy using fine needle aspirate (FNA) of primary lesions and/or peripheral blood samples. We enrolled a cohort of 35 patients referred for CT biopsy of primary pulmonary nodules, liver or adrenal masses and concurrently obtained FNA and peripheral blood samples. FNA sampling yielded sufficient material for μNMR analysis in 91% of cases and had a sensitivity and specificity of 91.6% and 100% respectively. Interestingly, among blood samples with positive circulating tumor cells (CTC), μNMR analysis of each patient's peripheral blood led to similar diagnosis (malignant vs benign) and differential diagnosis (lung malignancy subtype) in 100% and 90% (18/20) of samples, respectively. μNMR appears to be a valuable, non-invasive adjunct in the diagnosis of lung cancer.From the Clinical EditorThe authors of this study established that recently developed micro-nuclear magnetic resonance (μNMR) technology can be leveraged for diagnosing pulmonary malignancy using fine needle aspirate (FNA) of primary lesions and/or peripheral blood samples derived from 35 patients, suggesting practical clinical applicability of this technique.     

Monitoring circulating tumor cells in cancer vaccine trials

The presence of circulating tumor cells (CTC) from various cancers has provided a wealth of information and possibilities. As the role of CTC detection in the treatment assessment of metastatic breast cancer becomes standard, there is interest in applying this tool in cancer vaccine development and clinical trial monitoring. Since we lack a proven immunologic assay that correlates with clinical response, CTC detection, quantification and phenotypic characterization may be a useful surrogate for clinical outcome. The Cancer Vaccine Development Program is involved in the development of HER2/neu peptide based vaccine development for the prevention of recurrence in HER2/neu expressing cancers like breast cancer. The CellSearch System (Veridex, LLC Warren, NJ) has been used by our lab in conjunction with in vivo and/or in vitro immunologic measurements to define a monitoring tool that could predict clinical response. Once validated, this assay could significantly shorten clinical trials and lead to more efficient assessment of potentially promising cancer vaccines.     

磁共振动态增强结合血循环肿瘤细胞、循环游离 DNA对乳腺癌的临床诊断研究

目的探讨磁共振动态增强(DCE-MRI)结合血循环肿瘤细胞(CTCs)、循环游离 DNA(cfDNA)对乳腺癌的诊断价值。方法选取沧州市人民医院 2020年6月至 2021年11月符合纳入标准的乳腺癌病人 82例(乳腺癌组)。另选同期 65例乳腺良性病变且未合并其他疾病者作为良性组。比较两组 DCE-MRI定量参数、 CTCs、cfDNA水平,对比 MRI特征,分析 DCE-MRI结合 CTCs、cfDNA诊断乳腺癌的价值。结果乳腺癌组 DCE-MRI定量参数速度常数( Ktrans)值、速率常数( Kep)值、 Alu247/115水平及 CTCs计数分别为(0.18±0.04)min、(1.32±0.27)min、(12.97±2.14)个/毫升、 0.97±0.32,比良性组高(0.05±0.01)min、(0.51±0.11)min、(10.15±1.45)个/毫升、 0.55±0.12(P<0.05)。良性组边缘多光滑,形状规则,内部强化以均匀为主,时间 -信号强度曲线(TIC)以Ⅰ型多见,早期增强率 < 60%;乳腺癌组边缘毛刺征、形状不规则,内部不均匀强化为主,以TIC分型 Ⅲ型多见,早期增强率 <60%占比高。两组 MRI形态表现比较差异有统计学意义( P<0.05)。淋巴结转移、 Ⅲ~Ⅳ期者 CTCs数量、 Alu247/115水平明显高于无淋巴结转移、 Ⅰ~Ⅱ期者( P<     

TiO2 nanoparticles and bulk material stimulate human peripheral blood mononuclear cells

Nanomaterials are increasingly produced and used throughout recent years. Consequently the probability of exposure to nanoparticles has risen. Because of their small 1–100 nm size, the physicochemical properties of nanomaterials may differ from standard bulk materials and may pose a threat to human health. Only little is known about the effects of nanoparticles on the human immune system. In this study, we investigated the effects of TiO₂ nanoparticles and bulk material in the in vitro model of human peripheral blood mononuclear cells (PBMC) and cytokine-induced neopterin formation and tryptophan breakdown was monitored. Both biochemical processes are closely related to the course of diseases like infections, atherogenesis and neurodegeneration. OCTi60 (25 nm diameter) TiO₂ nanoparticles and bulk material increased neopterin production in unstimulated PBMC and stimulated cells significantly, the effects were stronger for OCTi60 compared to bulk material, while P25 TiO₂ (25 nm diameter) nanoparticles had only little influence. No effect of TiO₂ nanoparticles on tryptophan breakdown was detected in unstimulated cells, whereas in stimulated cells, IDO activity and IFN-γ production were suppressed but only at the highest concentrations tested. Because neopterin was stimulated and tryptophan breakdown was suppressed in parallel, data suggests that the total effect of particles would be strongly pro-inflammatory.     

循环肿瘤细胞在胃癌中应用的研究进展

复发和转移是导致胃癌术后低生存率的主要原因。循环肿瘤细胞在胃癌转移中的起到关键作用，所谓的“肿瘤细胞播散的上皮 -间质 /间质 -上皮假说”很好地解释了这一点。近年来，随着循环肿瘤细胞检测技术的进步，循环肿瘤细胞作为推动精准医学的重要技术得到了广泛的关注。该研究就循环肿瘤细胞在胃癌的早期诊断、治疗过程中的动态监测、预后的判断、个体化治疗中的研究进展及展望作一综述。     

Identification of serum biomarkers for lung cancer using magnetic bead-based SELDI-TOF-MS

Aim:

To identify novel serum biomarkers for lung cancer diagnosis using magnetic bead-based surface-enhanced laser desorption/ionization time-of-flight mass spectrum (SELDI-TOF-MS).

Methods:

The protein fractions of 121 serum specimens from 30 lung cancer patients, 30 pulmonary tuberculosis patients and 33 healthy controls were enriched using WCX magnetic beads and subjected to SELDI-TOF-MS. The spectra were analyzed using Bio-marker Wizard version 3.1.0 and Biomarker Patterns Software version 5.0. A diagnostic model was constructed with the marker proteins using a linear discrimination analysis method. The validity of this model was tested in a blind test set consisted of 8 randomly selected lung cancer patients, 10 pulmonary tuberculosis patients and 10 healthy volunteers.

Results:

Seventeen m/z peaks were identified, which were significantly different between the lung cancer group and the control (tuberculosis and healthy control) groups. Among these peaks, the 6445, 9725, 11705, and 15126 m/z peaks were selected by the Biomarker Pattern Software to construct a diagnostic model for lung cancer. This four-peak model established in the training set could discriminate lung cancer patients from non-cancer patients with a sensitivity of 93.3% (28/30) and a specificity of 90.5% (57/63). The diagnostic model showed a high sensitivity (75.0%) and a high specificity (95%) in the blind test validation. Database searching and literature mining indicated that the featured 4 peaks represented chaperonin (M9725), hemoglobin subunit beta (M15335), serum amyloid A ({"type":"entrez-nucleotide","attrs":{"text":"M11548","term_id":"340238","term_text":"M11548"}}M11548), and an unknown protein.

Conclusion:

A lung cancer diagnostic model based on bead-based SELDI-TOF-MS has been established for the early diagnosis or differential diagnosis of lung cancers.     

Specific targeting of cancer cells by multifunctional mitoxantrone-conjugated magnetic nanoparticles

Abstract

We report on the synthesis of bifunctional mitoxantrone (MTX)-grafted magnetic nanoparticles (MNPs) modified by dopamine-polyethylene glycol-folic acid (DPA-PEG-FA) for targeted imaging and therapy of cancer. MNPs (～7–10?nm) were synthesized using the thermal decomposition reaction of Fe(acac)₃. Bromoacetyl (BrAc) terminal polyethylene glycol dopamine (DPA-PEG-BrAc) was synthesized and treated with ethylene diamine to form bifunctional PEG moiety containing dopamine at one end and amino group at the other end (i.e. DPA-PEG-NH₂). It was then reacted with Fe₃O₄ nanoparticles (NPs) to form Fe₃O₄-DPA-PEG-NH₂ NPs. The activated folic acid (FA) was chemically coupled to Fe₃O₄-DPA-PEG-NH₂, forming Fe₃O₄-DPA-PEG-FA. MTX was then conjugated to Fe₃O₄-DPA-PEG-FA, forming Fe₃O₄-DPA-PEG-FA-MTX. Physicochemical characteristics of the engineered MNPs were determined. The particle size analysis and electron microscopy showed an average size of ～35?nm for Fe₃O₄-DPA-PEG-FA-MTX NPs with superparamagnetic behavior. FT-IR spectrophotometry analysis confirmed the conjugation of FA and MTX onto the MNPs. Fluorescence microscopy, cytotoxicity assay and flow cytometry analysis revealed that the engineered Fe₃O₄-DPA-PEG-FA-MTX NPs were able to specifically bind to and significantly inhibit the folate receptor (FR)-positive MCF-7 cells, but not the FR-negative A549 cells. Based upon these findings, we suggest the Fe₃O₄-DPA-PEG-FA-MTX NPs as an effective multifunctional-targeted nanomedicine toward simultaneous imaging and therapy of FR-positive cancers.     

Detection of circulating tumor cells in breast cancer with a refined immunomagnetic nanoparticle enriched assay and nested-RT-PCR

Early detection of circulation tumor cells (CTCs) in breast cancer patients has great clinical relevance. Currently, immunomagnetic microparticles enriched assays require Fe₃O₄ inner cores, making it difficult to improve sensitivity. In this study, we prepared magnetic nanoparticles with carbon-coated pure iron (Fe@C) acted as the core, Conjugating with EpCAM monoclonal antibody for immunomagnetic nanoparticles(IMPs). IMPs were used in conjunction with immunocytochemistry (ICC) to develop a refined immunomagnetic nanoparticles enriched assay (IMPEA) for detection of circulating tumor cells (CTCs) in breast cancer patients. Compared with nested RT-PCR, this method achieved the same sensitivity, but with a significantly reduced false-positive rate. This method will help find hidden micrometastases, establish clinical stage, and guide individual treatment post-surgery, suggesting potentially significant value in the clinic.From the Clinical EditorThis team of investigators prepared magnetic nanoparticles with carbon-coated pure iron as core and conjugated them with EpCAM monoclonal antibody to form immunomagnetic nanoparticles for circulating tumor cell (CTC) detection. Compared with nested RT-PCR, this method achieved the same sensitivity, but with a significantly reduced false-positive rate, paving the way to the development of a tool that enables enhanced detection of micrometastases and post-surgical treatment monitoring.     

循环肿瘤细胞对结直肠癌预后的相关性研究

目的 研究分析肥大细胞三种亚型对结直肠癌预后可能产生的影响,从而发现肥大细胞对结直肠癌预后的影响因素,为临床指导治疗提供理论依据.方法 选择2011年资料完整的结直肠癌根治术后石蜡标本40例作为试验组,术前未放疗或化疗;另取20例正常大肠组织,作为对照组,采用双重免疫组化技术对试验组做连续切片进行染色,观察2组中各亚型构成比是否有差异.计算出40例结直肠癌患者切片中各亚型的总平均计数值,与试验组每张切片相对应的平均计数值进行比较,观察肥大细胞各亚型的数量与性别、癌的分化程度、癌的浸润深度、淋巴结转移及生存时间的关系,并总结肥大细胞三种亚型与结直肠癌之间的关系.结果 TIM在结直肠癌根治术后石蜡标本有不同程度的浸润,2组的在结直肠癌的N分期和TNM分期及肿瘤分化程度方面差异有统计学意义(P<0.05).高计数组和低计数组5年总生存率和无瘤生存率分别为63.1%、59.3%和82.9%、79.0%,2组比较差异有统计学意义(P<0.05).COX比例风险模型单因素分析结果显示,在肿瘤组织中,肥大细的浸润是对结直肠癌患者总生存时间和无瘤生存时间造成影响的不良因素,后续多因素分析得出,对结直肠癌患者总生存时间、无瘤生存时间而言,浸润肥大细胞为其独立影响因素(P<0.05).结论 TIM与结直肠癌的N分期和TNM分期相关,循环肿瘤细胞可影响结直肠癌患者预后,所以可借助对循环肿瘤细胞的检测,去对结直肠癌患者的生存预后进行预测,提高患者的预后及生存率.     

Toxicity of cerium oxide nanoparticles in human lung cancer cells

With the fast development of nanotechnology, the nanomaterials start to cause people's attention for potential toxic effect. In this paper, the cytotoxicity and oxidative stress caused by 20-nm cerium oxide (CeO2) nanoparticles in cultured human lung cancer cells was investigated. The sulforhodamine B method was employed to assess cell viability after exposure to 3.5, 10.5, and 23.3 microg/ml of CeO2 nanoparticles for 24, 48, and 72 h. Cell viability decreased significantly as a function of nanoparticle dose and exposure time. Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species, glutathione, malondialdehyde, alpha-tocopherol, and lactate dehydrogenase, were quantitatively assessed. It is concluded from the results that free radicals generated by exposure to 3.5 to 23.3 microg/ml CeO2 nanoparticles produce significant oxidative stress in the cells, as reflected by reduced glutathione and alpha-tocopherol levels; the toxic effects of CeO2 nanoparticles are dose dependent and time dependent; elevated oxidative stress increases the production of malondialdehyde and lactate dehydrogenase, which are indicators of lipid peroxidation and cell membrane damage, respectively.     

循环肿瘤细胞在非小细胞肺癌患者中的水平及其与肿瘤标志物的关系研究

目的:探讨循环肿瘤细胞（CTCs）在非小细胞肺癌（NSCLC）患者中的水平及其与肿瘤标志物的关系,以为临床防治NSCLC提供一定的依据。方法:回顾性选取2020年3月至2021年2月江门市中心医院收治NSCLC患者50例作为研究对象,男29例、女21例,<60岁23例、≥60岁27例。并整理与分析其临床资料,所有患者均...     

Nanoresonator chip-based RNA sensor strategy for detection of circulating tumor cells: response using PCA3 as a prostate cancer marker

There is widespread interest in circulating tumor cells (CTCs) in blood. Direct detection of CTCs (often < 1/mL) is complicated by a number of factors, but the presence of ～10(3) to 10(4) copies of target RNA per CTC, coupled with simple enrichments, can greatly increase detection capability. In this study we used resonance frequency shifts induced by mass-amplifying gold nanoparticles to detect a hybridization sandwich bound to functionalized nanowires. We selected PCA3 RNA as a marker for prostate cancer, optimized antisense binding sites, and defined conditions allowing single nucleotide mismatch discrimination, and used a hybrid resonator integration scheme, which combines elements of top-down fabrication with strengths of bottom-up fabrication, with a view to enable multiplexed sensing. Bound mass calculated from frequency shifts matched mass estimated by counting gold nanoparticles. This represents the first demonstration of use of such nanoresonators, which show promise of both excellent specificity and quantitative sensitivity.     

Pharmacological profiling of disulfiram using human tumor cell lines and human tumor cells from patients

The thiocarbamate drug disulfiram has been used for decades in the treatment of alcohol abuse. Disulfiram induces apoptosis in a number of tumor cell lines and was recently by us proposed to act as a 26S proteasome inhibitor. In this work we characterized disulfiram in vitro with regard to tumor-type specificity, possible mechanisms of action and drug resistance and cell death in human tumor cell lines and in 78 samples of tumor cells from patients using the fluorometric microculture cytotoxicity assay and the automated fluorescence-imaging microscope ArrayScan((R)). Disulfiram induced cytotoxicity in a biphasic pattern in both cell lines and patient tumor cells. Disulfiram induced apoptosis as measured by cell membrane permeability, nuclear fragmentation/condensation and caspase-3/7 activation using high content screening assays. For many of the cell lines tested disulfiram was active in sub-micromolar concentrations. When comparing the logIC(50) patterns with other cytotoxic agents, disulfiram showed low correlation (R<0.5) with all drugs except lactacystin (R=0.69), a known proteasome inhibitor, indicating that the two substances may share mechanistic pathways. Disulfiram was more active in hematological than in solid tumor samples, but substantial activity was observed in carcinomas of the ovary and the breast and in non-small cell lung cancer. Disulfiram also displayed higher cytotoxic effect in cells from chronic lymphocytic leukemia than in normal lymphocytes (p<0.05), which may indicate some tumor selectivity. These results together with large clinical experience and relatively mild side effects encourage clinical studies of disulfiram as an anti-cancer agent.     

In vitro toxicity of silica nanoparticles in human lung cancer cells

The cytotoxicity of 15-nm and 46-nm silica nanoparticles was investigated by using crystalline silica (Min-U-Sil 5) as a positive control in cultured human bronchoalveolar carcinoma-derived cells. Exposure to 15-nm or 46-nm SiO(2) nanoparticles for 48 h at dosage levels between 10 and 100 microg/ml decreased cell viability in a dose-dependent manner. Both SiO(2) nanoparticles were more cytotoxic than Min-U-Sil 5; however, the cytotoxicities of 15-nm and 46-nm silica nanoparticles were not significantly different. The 15-nm SiO(2) nanoparticles were used to determine time-dependent cytotoxicity and oxidative stress responses. Cell viability decreased significantly as a function of both nanoparticle dosage (10-100 microg/ml) and exposure time (24 h, 48 h, and 72 h). Indicators of oxidative stress and cytotoxicity, including total reactive oxygen species (ROS), glutathione, malondialdehyde, and lactate dehydrogenase, were quantitatively assessed. Exposure to SiO(2) nanoparticles increased ROS levels and reduced glutathione levels. The increased production of malondialdehyde and lactate dehydrogenase release from the cells indicated lipid peroxidation and membrane damage. In summary, exposure to SiO(2) nanoparticles results in a dose-dependent cytotoxicity in cultural human bronchoalveolar carcinoma-derived cells that is closely correlated to increased oxidative stress.     

肾癌外周血循环肿瘤细胞的筛查研究

目的探讨细胞角蛋白(CK)19作为循环肿瘤细胞的间接标志物的价值,以及CK19在肾癌患者外周血中的表达水平以及临床意义。方法选取健康人50名作为对照组,50例肾细胞癌患者作为肾癌组,首先进行对照组和肾癌组间CK19的指标测定,采用流式细胞术检测上述人群外周血中CK19的表达情况;再分析肾癌组内CK19的表达水平与肾癌病理分型、病理分期及组织学分级之间的关系。结果 50名对照组中无一例CK19阳性,50例肾癌组中24例CK19阳性;不同肾癌病理分型之间CK19表达差异无统计学意义(P>0.05),不同肾癌病理分期及组织学分级之间CK19表达差异具有统计学意义(P<0.05)。结论 CK19能够初步判断患者外周血循环肿瘤细胞的水平,对早期无创性诊断肾癌、预测其生物学行为有一定的参考价值,值得进一步探索和研究。     

Comparison of in vitro toxicity of silver ions and silver nanoparticles on human hepatoma cells

Scientific information on the potential harmful effects of silver nanoparticles (AgNPs) on human health severely lags behind their exponentially growing applications in consumer products. In assessing the toxic risk of AgNP usage, liver, as a detoxifying organ, is particularly important. The aim of this study was to explore the toxicity mechanisms of nano and ionic forms of silver on human hepatoblastoma (HepG2) cells. The results showed that silver ions and citrate‐coated AgNPs reduced cell viability in a dose‐dependent manner. The IC50 values of silver ions and citrate‐coated AgNPs were 0.5 and 50 mg L^?1, respectively. The LDH leakage and inhibition of albumin synthesis, along with decreased ALT activity, indicated that treatment with either AgNP or Ag ions resulted in membrane damage and reduced the cell function of human liver cells. Evaluation of oxidative stress markers demonstrating depletion of GSH, increased ROS production, and increased SOD activity, indicated that oxidative stress might contribute to the toxicity effects of nano and ionic forms of silver. The observed toxic effect of AgNP on HepG2 cells was substantially weaker than that caused by ionic silver, while the uptake of nano and ionic forms of silver by HepG2 cells was nearly the same. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 679–692, 2016.