第1期 | Countstar与科研之间的故事

发布日期:2020-12-31 15:32:03浏览次数:653

本周Countstar在应用发表的文献中选取了6篇分享给大家。第1篇是关于中草药药理方面,阐述了补骨脂素对于肝毒性的影响。第2篇文章介绍了多发性骨髓瘤的发病机理,骨髓瘤中非编码RNA DARS-AS1被缺氧诱导因子(HIF)-1直接调节,为我们理解多发性骨髓瘤提供了更多的机制基础。第3、4篇文章分别介绍了肺癌、结直肠癌各自癌细胞的调控机理。第5、6两篇文章是在新型的化学材料上进行了研究,为解决疾病提供新的辅助工具。感兴趣的文章可以去寻找一下原文。


1.The Accumulation of Psoralen Contributes to ItsHepatotoxicity Revealed by Pharmacokinetic and Toxicokinetic Study afterRepeated Administration

通过药代动力学和毒代动力学研究表明补骨脂素的积累是引起肝毒性的原因。


ABSTRACT: Psoralen is afuranocoumarin compound found in many herb medicines and is claimed tocontribute to the hepatotoxicity caused by lots of traditional Chinesemedicine. So far, there has been no research on the difffferences inpharmacokinetics of single and repeated dosing of psoralen. Moreover, theresearch on the cumulative toxicity of low concentration and long-termadministration on cells has not been reported. Therefore, this studyinvestigated the pharmacokinetic difffferences and the accumulated cytotoxicityof psoralen from repeated administration. The study found that after single orrepeated administration of psoralen for 3 months at various dosages (14, 28, and56 mg/kg), the pharmacokinetic parameters of female rats between single doseand repeated dose administration are totally difffferent. Compared with asingle administration, multiple administrations increased psoralen’s in vivoexposure, prolonged the peak time, prolonged the half-life of the drug, reducedthe drug clearance rate, and prolonged the drug’s stay in the body. HepG2 cellswere exposed to low doses (5, 10, 20, or 40 μM) of psoralen for 1, 2, 3, or 4days. A 20 and 40 μM dose of psoralen did not induced cell death in the 1st daybut signifificantly decreased the cell viability at the 3rd and 4th day ofrepeated administration, respectively. In addition, multiple administrations ofpsoralen decreased cell viability due to G2 arrest.

摘要:补骨脂素是呋喃香豆素的一种化合物,存在于许多草药中,据称可有助于许多中药引起肝毒性。到目前为止,还没有差异的研究。关于补骨脂素单次和重复给药,低浓度累积毒性的研究尚未进行。因此,本研究调查了补骨脂素药代动力学的差异和重复给药引起的细胞毒性。研究发现,单次或重复给药后剂量(14、28和56 mg / kg)补骨脂素治疗3个月,雌性大鼠单次之间的剂量和重复给药完全不同。与单次给药相比,多次给药增加了补骨脂素的体内暴露量,延长了峰值时间,延长了药物的半衰期,降低了药物清除率,延长了药物在体内的停留时间。HepG2细胞暴露于低剂量(5、10、20或40μM)补骨脂素中1、2、3或4天。补骨脂素20、40μM剂量在第1天没有诱导细胞死亡,但在重复给药的第3天和第4天显著降低细胞活力。



2.Hypoxia-induced long non-coding RNA DARS-AS1 regulates RBM39 stabilitytopromote myeloma

缺氧诱导RNA DARS-AS1调节RBM39稳定性促进骨髓瘤


ABSTRACT: Multiple myeloma is amalignant plasma-cell disease, which is highly dependent on the hypoxic bonemarrow microenvironment.However, the underlying mechanisms of hypoxiacontributing to myeloma genesis are not fully understood. Here, we show that longnoncoding RNA DARS-AS1 in myeloma is directly upregulated by hypoxia induciblefactor (HIF)-1. Importantly, DARS-AS1 is required for the survival andtumorigenesis of myeloma cells both in vitro and in vivo. DARS-AS1 exerts itsfunction by binding RNA-binding motif protein 39 (RBM39),which impedes theinteraction between RBM39 and its E3 ubiquitin ligase RNF147, and preventsRBM39 from degradation. The overexpression of RBM39 observed in myeloma cellsis associated with poor prognosis.Furthermore, knockdown of DARS-AS1 inhibitsthe mammalian target of rapamycin signaling pathway, an effect that is reversedby RBM39 overexpression. We reveal that a novel HIF-1/DARS-AS1/RBM39 pathway isimplicated in the pathogenesis of myeloma. Targeting DARS-AS1/RBM39 may,therefore,represent a novel strategy to combat myeloma.

摘要:多发性骨髓瘤是一种恶性细胞病,极易受到缺氧骨髓微环境的影响。然而,缺氧促进骨髓瘤发生的机制尚不完全清楚。在这里,我们发现骨髓瘤中非编码RNA DARS-AS1被缺氧诱导因子(HIF)-1直接调节。重要的是,DARS-AS1对骨髓瘤细胞的生存和体内肿瘤的产生都是必需的。DARS-AS1通过RNA结合蛋白39(RBM39)发挥其功能,从而阻碍RBM39及E3泛素连接酶RNF147的相互作用,并防止RBM39蛋白降解。骨髓瘤细胞中RBM39蛋白的过度表达与不良预后有关。此外,DARS-AS1的敲除抑制了雷帕霉素信号通路的靶点,这种作用被RBM39过度表达所逆转。我们发现一种新的HIF-1/DARS-AS1/RBM39通路参与了骨髓瘤的发病机制。因此,靶向DARS-AS1/RBM39可能是一种治疗骨髓瘤的新策略。



3.Long Non-coding RNA HOTAIR Function as a CompetingEndogenous RNA for miR-149-5p to Promote the Cell Growth, Migration,andInvasion in Non-small Cell Lung Cancer

非编码RNA HOTAIR作为miR-149-5p的竞争性内源RNA,促进非小细胞肺癌细胞的生长、迁移和侵袭



ABSTRACT: Lung cancer is a leadingcause of cancer death all around the world. Long non-coding RNAs (lncRNAs) havebeen confirmed to be involved in carcinogenesis of malignancies.However, themolecular mechanism of most lncRNAs in various kinds of cancers remainsunclear. LncRNA HOTAIR and HNRNPA1 are reported to play an oncogenic role innon-small cell lung cancer, and the overexpression of HNRNPA1 is shown topromote the proliferation of lung adenocarcinoma cells. In our study, we findthat the overexpression of HOTAIR could promote the proliferation and overexpressionof miR-149-5p could inhibit the proliferation of lung cancer cells. Flowcytometric analysis determines that overexpression of miR-149-5p induces cellcycle arrest in the G0/G1

phases, whereas overexpression ofHOTAIR decreases the proportion of G0/G1phase cells. Also, overexpression ofHOTAIR promotes the migration and invasion ability of lung cancer cells,confirmed by the wound-healing and transwell assays, which are suppressed byoverexpression of miR-149-5p. Furthermore, the dual-luciferase reporter assayindicates that miR-149-5p could bind both HOTAIR and the 3′UTR of HNRNPA1.Insummary, we find that HOTAIR can regulate HNRNPA1 expression through a ceRNAmechanism by sequester miR-149-5p, which post-transcriptionally targetsHNRNPA1,thus promoting lung cancer progression.

摘要:肺癌是全世界癌症死亡率的最高的疾病之一。非编码RNA(lncRNA)已被证实与恶性肿瘤的发生有关。然而,大多数lncRNAs在各种癌症中的分子机制还不清楚。据报道,LncRNA HOTAIR和HNRNPA1在非小细胞肺癌中具有致癌作用,HNRNPA1的过度表达促进了肺腺癌细胞的增殖。在我们的研究中,我们发现HOTAIR的过度表达可以促进肺癌细胞的增殖,miR-149-5p的过度表达可以抑制肺癌细胞的增殖。流式细胞仪分析确定miR-149-5p的过度表达可诱导细胞周期停滞在G0/G1期,而HOTAIR的过度表达则会降低G0/G1期细胞的比例。另外,HOTAIR的过度表达促进了肺癌细胞的迁移和侵袭能力,miR-149-5p的过度表达抑制了这种迁移和侵袭能力。此外,双荧光素酶报告试验表明miR-149-5p可以结合HOTAIR和HNRNPA1的3′UTR,我们发现HOTAIR可以通过ceRNA机制调节HNRNPA1的表达,miR-149-5p在转录后靶向HNRNPA1,从而促进肺癌的进展。

4.Transcription Factor EBF1 Over-ExpressionSuppresses TumorGrowth in vivo and in vitro via Modulation of the PNO1/p53Pathwayin Colorectal Cancer

通过转录因子EBF1过度表达调控PNO1/p53通路抑制结直肠癌的体内外肿瘤生长


ABSTRACT: Early B cell factor 1(EBF1) has been identified as an upstream transcription factor of the potentialoncogene PNO1 and is involved in the growth of colorectal cancer (CRC) cells.However, its expression, biological function, and underlying mechanism ofaction in most solid tumors remain largely unknown. We postulated that EBF1 hasa role in the pathophysiology of CRC. Analysis of EBF1 mRNA expression in CRCtumor samples from several public databases and directly from banked tissuesrevealed that EBF1 mRNA expression is lower in CRC tissue compared tonon-cancerous colorectal tissue. Survival analysis of multiple datasetsrevealed that low EBF1 expression was correlated with shorter overall survival,relapse-free survival, and event-free survival in CRC patients. Transduction oflentivirus encoding full length EBF1 followed by in vitro and in vivo assaysdemonstrated that EBF1 over-expression in CRC cell lines suppresses cell growthby inhibiting cell viability, cell survival, and induces cell cycle arrest andapoptosis. Mechanistic investigation indicated that EBF1 over-expressiondown-regulates PNO1 mRNA and protein expression, as well as transcriptionalactivity while up-regulating the expression of p53 and p21 proteins. Thesefindings suggest that EBF1 is a novel potential tumor suppressor in CRC withprognostic value for the identification of patients at high-risk of relapse.

摘要:早期B细胞因子(EBF1)已被鉴定为潜在致癌基因PNO1的上游转录因子,并参与了结直肠癌(CRC)细胞的生长。然而,其在大多数实体瘤中的表达、生物学功能和潜在作用机制尚不清楚。我们假设EBF1在CRC的病理中起作用。对来自多个公共数据库和直接来自组织的CRC肿瘤样品中EBF1 mRNA的分析表明,与非癌性结直肠组织相比,CRC组织中EBF1 mRNA表达更低。多个数据的分析表明,CRC患者中低EBF1表达与较短的总生存期,无复发生存期和无事件生存期相关。通过转染EBF1的慢病毒,然后在体外和体内试验表明,CRC细胞系中EBF1的过量表达可抑制细胞通过抑制细胞活力,通过诱导细胞周期停滞和凋亡来抑制细胞的生长。机机制研究表明,EBF1过度表达下调pno1mrna和蛋白的表达,以及转录活性,同时上调p53和p21蛋白的表达。这些发现提示EBF1是一种新的潜在的大肠癌抑癌因子,对鉴别复发高危患者具有预后价值。


5.Immunostimulatory Potential of MoS2 Nanosheets:Enhancing Dendritic Cell Maturation,
Migration and T Cell Elicitation /MoS2纳米片的免疫刺激潜力:增强树突状细胞成熟,迁移和T细胞诱导。

ABSTRACT: Due to their extraordinaryphysical and chemical properties, MoS2 nanosheets (MSNs) are becoming morewidely used in nanomedicine. However, their influence on immune systems remainsunclear.Two few-layered MSNs at sizes of 100–250 nm (S-MSNs) and 400–500 nm(L-MSNs) were used in this study. Bone marrow-derived dendritic cells (DCs)were exposed to both MSNs at different doses (0, 8, 16, 32, 64, 128 µg/mL) for48 h and subjected to analyses of surface marker expression, cytokinesecretion, lymphoid homing and in vivo T cell priming.Few-layered MSNs rangingfrom 100 to 500 nm in size could play an immunostimulatory role in enhancing DCmaturation, migration and T cell elicitation,making them a good candidate forvaccine adjuvants. Investigation of this study will not only expand theapplications of MSNs and other new transition metal dichalcogenides (TMDCs) butalso shed light on the in vivo immune-risk evaluation of MSN-basednanomaterials.

摘要:由于其特殊的物理化学性质,MoS2纳米片在纳米医学中的应用越来越广泛。然而,它们对免疫系统的影响仍不清楚。本研究使用了两种尺寸分别为100-250nm(S-MSNs)和400-500nm(L-MSNs)的多层MSNs。将骨髓源性树突状细胞(DC)以不同剂量(0、8、16、32、64、128μg/mL)暴露于两种MSN中48h,并进行表面标记物表达、细胞因子分泌、淋巴归巢和体内T细胞的分析。粒径在100~500nm之间的多层MSN在促进DC成熟、迁移和T细胞诱导方面具有免疫刺激作用,使其成为疫苗佐剂的良好候选者。本研究的开展不仅拓展了MSNs等新型TMDC的应用范围,而且对MSN纳米材料的体内免疫风险评估具有重要意义。


6.Encapsulation and release of living tumor cellsusing hydrogels with the hybridization chain reaction


ABSTRACT: Abstract Circulating tumor cells(CTCs) enable noninvasive liquid biopsy and identification of cancer. Variousapproaches exist for the capture and release of CTCs, including microfluidicmethods and those involving magnetic beads or nanostructured solid interfaces.However, the concomitant cell damage and fragmentation that often occur duringcapture make it difficult to extensively characterize and analyze living CTCs.Here, we describe an aptamer-trigger-clamped hybridization chain reaction(atcHCR) method for the capture of CTCs by porous 3D DNA hydrogels. The 3Denvironment of the DNA networks minimizes cell damage, and the CTCs cansubsequently be released for live-cell analysis. In this protocol, initiatorDNAs with aptamer-toehold biblocks specifically bind to the epithelial celladhesion molecule (EpCAM) on the surface of CTCs, which triggers the atcHCR andthe formation of a DNA hydrogel. The DNA hydrogel cloaks the CTCs, facilitatingquantification with minimal cell damage. This method can be used toquantitively identify as few as 10 MCF-7 cells in a 2-µL blood sample.Decloaking of tumor cells via gentle chemical stimulus (ATP) is used to releaseliving tumor cells for subsequent cell culture and live-cell analysis. We alsodescribe how to use the protocol to encapsulate and release cells of cancercell lines, which can be used in preliminary experiments to model CTCs. Thewhole protocol takes ~2.5 d to complete, including downstream cell culture andanalysis.
摘要:循环肿瘤细胞(CTC)可以进行无创液体活检和癌症鉴定。捕获和释放CTC的方法多种多样,包括微流体方法以及涉及磁珠或纳米结构固体的方法。然而,捕获过程中经常发生的伴随细胞损伤和碎片化使得难以广泛统计和分析活的CTC。在这里,我们描述了一种使用多孔三维DNA水凝胶的atcHCR方法捕获CTCs。DNA网络的3D环境最大程度地减少了细胞损伤,随后可以释放CTC进行活细胞分析。在该方案中,带有toehold biblocks的起始DNA与CTCs表面的上皮细胞粘附分子(EpCAM)特异性结合,从而触发atcHCR并形成DNA水凝胶。DNA水凝胶保护CTC,使细胞损伤最小,便于量化。该方法可用于定量鉴定2 µL血液样本中的多达10个MCF-7细胞。通过温和的化学刺激(ATP)使肿瘤细胞受损,以释放活的肿瘤细胞,用于随后的细胞培养和活细胞分析。我们还描述了如何使用该方案来封装和释放癌细胞系的细胞,可将其用于初步实验中以模拟CTC。整个方案需要约2.5 d的时间才能完成,包括下游细胞培养和分析。


Countstar Fluorescence Cell Analyzer