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5′-tiRNA-Gln inhibits hepatocellular carcinoma progression by repressing translation through the interaction with eukaryotic initiation factor 4A-I

《医学前沿（英文）》 2023年第17卷第3期页码 476-492 doi: 10.1007/s11684-022-0966-6

摘要： tRNA-derived small RNAs (tsRNAs) are novel non-coding RNAs that are involved in the occurrence and progression of diverse diseases. However, their exact presence and function in hepatocellular carcinoma (HCC) remain unclear. Here, differentially expressed tsRNAs in HCC were profiled. A novel tsRNA, tRNA^Gln-TTG derived 5′-tiRNA-Gln, is significantly downregulated, and its expression level is correlated with progression in patients. In HCC cells, 5′-tiRNA-Gln overexpression impaired the proliferation, migration, and invasion in vitro and in vivo, while 5′-tiRNA-Gln knockdown yielded opposite results. 5′-tiRNA-Gln exerted its function by binding eukaryotic initiation factor 4A-I (EIF4A1), which unwinds complex RNA secondary structures during translation initiation, causing the partial inhibition of translation. The suppressed downregulated proteins include ARAF, MEK1/2 and STAT3, causing the impaired signaling pathway related to HCC progression. Furthermore, based on the construction of a mutant 5′-tiRNA-Gln, the sequence of forming intramolecular G-quadruplex structure is crucial for 5′-tiRNA-Gln to strongly bind EIF4A1 and repress translation. Clinically, 5′-tiRNA-Gln expression level is negatively correlated with ARAF, MEK1/2, and STAT3 in HCC tissues. Collectively, these findings reveal that 5′-tiRNA-Gln interacts with EIF4A1 to reduce related mRNA binding through the intramolecular G-quadruplex structure, and this process partially inhibits translation and HCC progression.

关键词： EIF4A1 G-quadruplex hepatocellular carcinoma tRNA-derived small RNA translation initiation

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Paternal environmental exposure-induced spermatozoal small noncoding RNA alteration meditates the intergenerational

《医学前沿（英文）》 2022年第16卷第2期页码 176-184 doi: 10.1007/s11684-021-0885-y

摘要： Studies of human and mammalian have revealed that environmental exposure can affect paternal health conditions as well as those of the offspring. However, studies that explore the mechanisms that meditate this transmission are rare. Recently, small noncoding RNAs (sncRNAs) in sperm have seemed crucial to this transmission due to their alteration in sperm in response to environmental exposure, and the methodology of microinjection of isolated total RNA or sncRNAs or synthetically identified sncRNAs gradually lifted the veil of sncRNA regulation during intergenerational inheritance along the male line. Hence, by reviewing relevant literature, this study intends to answer the following research concepts: (1) paternal environmental factors that can be passed on to offspring and are attributed to spermatozoal sncRNAs, (2) potential role of paternal spermatozoal sncRNAs during the intergenerational inheritance process, and (3) the potential mechanism by which spermatozoal sncRNAs meditate intergenerational inheritance. In summary, increased attention highlights the hidden wonder of spermatozoal sncRNAs during intergenerational inheritance. Therefore, in the future, more studies should focus on the origin of RNA alteration, the target of RNA regulation, and how sncRNA regulation during embryonic development can be sustained even in adult offspring.

关键词： small noncoding RNAs epigenetic inheritance paternal intergenerational inherence extracellular vesicles

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Blockage of receptor-interacting protein 2 expression by small interfering RNA in murine macrophages

LIU Hongchun, CAO Zhongwei, JIN Jianjun, WANG Jiyao

《医学前沿（英文）》 2008年第2卷第2期页码 166-170 doi: 10.1007/s11684-008-0030-1

摘要： This study aims to demonstrate that blocking the receptor-interacting protein2 (Rip2) expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality. Murine Rip2 small interfering RNA (siRNA) plasmids were constructed and transfected into macrophage and Rip2 expression was detected with reverse transcription-polymerase chain reaction (RT-PCR) and western blot. Cell proliferation was assayed with MTT. TNF-? concentration was assayed with ELISA and high-mobility group box 1 protein (HMGB1) level with semi-quantitative western blot after lipopolysaccharide (LPS) stimulation. LPS challenge was given after the plasmids were injected into mice and the survival rate was calculated. Rip2 siRNA plasmid could block the mRNA and protein expression of Rip2 and promote cell proliferation. Blocking Rip2 could attenuate LPS-induced TNF-? and HMGB1 production. The HMGB1 expression in the liver decreased to (40.21 ± 11.03) pg/g, and serum TNF-? level decreased to (300.43 ± 59.26) ng/L ( < 0.05). The survival rate of mice from endotoxemia was also improved ( < 0.05). The results demonstrate that Rip2 siRNA plasmid can block the expression of Rip2, decrease the production of TNF-? and HMGB1 and protect mice from fatal endotoxemia.

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Gene silencing efficiency of shRNA expression vectors targeting Cx43

Cuihong ZHENG, Yunxia WU, Guangying HUANG, Wei WANG

《医学前沿（英文）》 2009年第3卷第2期页码 130-135 doi: 10.1007/s11684-009-0030-9

摘要： Our previous studies showed that there were close relationships between connexin 43 (Cx43) and acupoints and meridians. In order to further investigate the effect of Cx43 in acupuncture treatment, RNA interference technique was used to construct small hairpin RNA (shRNA) expression vectors targeting Cx43 and identify the efficiency of RNA interference in NIH/3T3 cell lines for further use . Aiming directly at the two targets of Cx43 mRNA sequence of the rat and mouse homology region, we synthesized two pairs of complementary oligonucleotide strands . Double strands were formed after annealing, and then inserted into Pgenesil-1 plasmid expression vector. After identification by enzyme cutting and sequencing, the recombinant plasmids named P-Cx43-shRNA (1), P-Cx43-shRNA (2) and P-con-shRNA were transfected into the NIH/3T3 cells. Immunofluorescence and Western blot assays were used to detect the protein level of Cx43 after being screened by G418.The results of enzyme cutting and sequencing showed that we successfully constructed two shRNA expression vectors targeting Cx43, and a control expression vector for rat and mouse. Also, the Cx43 protein level was decreased by 73.5% ( < 0.01) and 10.8%, accordingly. The Cx43 protein level was not influenced by the transfection of P-con-shRNA. The outcomes demonstrate that the plasmid P-Cx43-shRNA (1) can specifically silence better the expression of Cx43 in NIH/3T3 cells, which offers an experimental evidence for further investigation.

关键词： RNA interference connexin 43 small hairpin RNA (shRNA) acupuncture

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肿瘤特异性环状RNA来源抗原肽被证实存在于肝胆肿瘤中 Article

王文闻, 马莉丽 , 邢政, 苑廷淦, 包金霞, 朱妍静, 赵晓芳, 赵燕, 宗雅丽, 张雅妮, 莘似韵, 邱辛瑶, 杨帅, 王红阳, 高栋, 王鹏, 陈磊

《工程（英文）》 2023年第22卷第3期页码 159-170 doi: 10.1016/j.eng.2022.06.008

摘要：在本研究中，利用肝胆肿瘤类器官分析了环状RNA（circRNA）作为肿瘤抗原肽新来源的潜力。使用RNA测序（RNA-seq）和基于算法的评分工具，预测出27 个类器官中3950 个肿瘤特异性circRNA 可翻译18 971 条抗原肽。

关键词：肿瘤抗原患者来源肝胆肿瘤类器官环状RNA 基于质谱的免疫肽组学

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Ribozyme and the mechanisms that underlie RNA catalysis

Timothy J. Wilson,Yijin Liu,David M. J. Lilley

《化学科学与工程前沿（英文）》 2016年第10卷第2期页码 178-185 doi: 10.1007/s11705-016-1558-2

摘要： Ribozymes are widespread, and catalyze some extremely important reactions in the cell. Mechanistically most fall into one of two classes, using either metal ions or general acid-base catalysis. The nucleolytic ribozymes fall into the latter class, mostly using nucleobases. A sub-set of these use a combination of guanine base plus adenine acid to catalyze the cleavage reaction. New ribozymes are still being discovered at regular intervals and we can speculate on the potential existence of ribozymes that catalyze chemistry beyond phosphoryl transfer reactions, perhaps using small-molecule coenzymes.

关键词： RNA catalysis RNA structure catalytic mechanism

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抗RNA病毒相关生物材料

姚康德,尹玉姬,张宝连,赵立国

《中国工程科学》 2003年第5卷第7期页码 17-23

摘要：

在介绍RNA病毒结构、生殖、复制和转录的基础上，综述了抗病毒策略，其中包括抗SARS药物设计，RNA干扰，DNA疫苗释放系统，调控蛋白与糖胺聚糖衍生物或类似物相互作用，天然药物及肺泡组织工程等。这些实例涵盖了RNA病毒与蛋白质、DNA及多糖等生物材料的相互作用。生物材料作为基质或载体正在向细胞或/和基因活化的第三代生物材料发展，可望在抗病毒中发挥作用。

关键词：抗RNA病毒蛋白质 DNA 多糖生物材料

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基于RNA的生物防治——一种作物保护新模式 Review

Matthew Bramlett, Geert Plaetinck, Peter Maienfisch

《工程（英文）》 2020年第6卷第5期页码 522-527 doi: 10.1016/j.eng.2019.09.008

摘要：在过去的几年中，RNA干扰（RNAi）过程被认为是一种非常有前景的新方法，可作为化学和生物害虫防治剂、植物保护剂等叶面喷施、土壤或种子处理的补充。基于RNA的活性成分（AI）具有独特的作用方式，可以通过基因修饰（GM）和生物防治两种途径来实现。由于基于RNA的AI可利用自然过程来发挥控制作用，同时它们具有高度选择性，降低了非目标生物（NTO）的风险，因此基于RNA的AI有望提供未来作物保护剂所需要的选择性和可持续性。本文讨论了基于RNA的生物防治的替代方案在作物保护中的优势和局限性，以及RNA生物防治科罗拉多马铃薯甲虫（CPB）、玉米根虫（CRW）和大豆臭虫（SSB）的最新研究进展。在实现各种基于RNA的产品及其广泛使用和应用的道路上，仍然存在许多挑战。尽管如此，我们仍可预期到，基于RNA的AI将成为有价值的新工具，以补充当前的农作物保护解决方案。

关键词：基于RNA的生物防治 RNA干扰（RNAi）科罗拉多马铃薯甲虫（CPB）玉米根虫（CRW）大豆臭虫（SSB）

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岩浆镍铜铂族矿床成矿过程中流体的作用——对小岩体超大型矿床的启示

张铭杰,汤庆艳,李文渊、余明、胡沛青、李建平

《中国工程科学》 2015年第17卷第2期页码 40-49

摘要：

超大型岩浆镍、铂族元素（PGE）硫化物矿床控制了Ni-PGE的资源量，本文根据地幔岩浆事件中Ni、Cu和PGE等在部分熔融、岩浆结晶和硫化物熔离过程的行为，讨论了岩浆Ni-Cu-PGE硫化物矿床的成矿岩浆类型与规模、成矿金属元素聚集的方式、岩浆演化过程中硫饱和度与硫化物液相不混溶的控制因素，探讨了流体对成矿金属的运移聚集作用等。认为地幔大规模的高度部分熔融（即要求高Mg岩浆）促使地幔硫化物及橄榄石中巨量的Ni和PGE进入岩浆，硫化物熔离富集成矿金属形成大型层状（或通道）岩体赋存的超大型Ni-Cu-PGE硫化物矿床；地幔超临界流体可能大范围内搬运聚集大量Ni-PGE金属元素，对岩浆硫化物矿床成矿可能有重要贡献，是小规模岩浆（小岩体）成矿的可能机制。

关键词：流体；小岩体；成矿作用

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敲低特异性环状非编码RNA显著抑制骨肉瘤的进展 Article

王世东, 张红亮, 李博, 陈成龙, 任婷婷, 黄怡, 刘凯, 李敬敬, 郭卫

《工程（英文）》 2023年第21卷第2期页码 187-193 doi: 10.1016/j.eng.2021.12.007

摘要：异常表达的非编码环状RNA（circRNA）对骨肉瘤的发生和发展至关重要。本研究的目的是探索一种新的circRNA circ_000203 在骨肉瘤中的表达和作用，阐明其潜在机制。

关键词：非编码RNA 骨肉瘤 circRNA 分子机制敲低

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Overexpressed long noncoding RNA CRNDE with distinct alternatively spliced isoforms in multiple cancers

Xuefei Ma, Wei Zhang, Rong Zhang, Jingming Li, Shufen Li, Yunlin Ma, Wen Jin, Kankan Wang

《医学前沿（英文）》 2019年第13卷第3期页码 330-343 doi: 10.1007/s11684-017-0557-0

摘要： Alternative splicing is a tightly regulated process that contributes to cancer development. CRNDE is a long noncoding RNA with alternative splicing and is implicated in the pathogenesis of several cancers. However, whether deregulated expression of CRNDE is common and which isoforms are mainly involved in cancers remain unclear. In this study, we report that CRNDE is aberrantly expressed in the majority of solid and hematopoietic malignancies. The investigation of CRNDE expression in normal samples revealed that CRNDE was expressed in a tissue- and cell-specific manner. Further comparison of CRNDE expression in 2938 patient samples from 15 solid and hematopoietic tumors showed that CRNDE was significantly overexpressed in 11 malignancies, including 3 reported and 8 unreported, and also implicated that the overexpressed isoforms differed in various cancer types. Furthermore, anti-cancer drugs could efficiently repress CRNDE overexpression in cancer cell lines and primary samples, and even had different impacts on the expression of CRNDE isoforms. Finally, experimental profiles of 12 alternatively spliced isoforms demonstrated that the spliced variant CRNDE-g was the most highly expressed isoform in multiple cancer types. Collectively, our results emphasize the cancer-associated feature of CRNDE and its spliced isoforms, and may provide promising targets for cancer diagnosis and therapy.

关键词： long noncoding RNA CRNDE alternative splicing

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Association of gene variants with juvenile amyotrophic lateral sclerosis

《医学前沿（英文）》 doi: 10.1007/s11684-023-1005-y

摘要： Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive degeneration of motor neurons, and it demonstrates high clinical heterogeneity and complex genetic architecture. A variation within TRMT2B (c.1356G>T; p.K452N) was identified to be associated with ALS in a family comprising two patients with juvenile ALS (JALS). Two missense variations and one splicing variation were identified in 10 patients with ALS in a cohort with 910 patients with ALS, and three more variants were identified in a public ALS database including 3317 patients with ALS. A decreased number of mitochondria, swollen mitochondria, lower expression of ND1, decreased mitochondrial complex I activities, lower mitochondrial aerobic respiration, and a high level of ROS were observed functionally in patient-originated lymphoblastoid cell lines and TRMT2B interfering HEK293 cells. Further, TRMT2B variations overexpression cells also displayed decreased ND1. In conclusion, a novel JALS-associated gene called TRMT2B was identified, thus broadening the clinical and genetic spectrum of ALS.

关键词： TRMT2B amyotrophic lateral sclerosis mitochondrial complex I tRNA methylation reactive oxygen species

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Potential use of serum HBV RNA in antiviral therapy for chronic hepatitis B in the era of nucleos(t)ide

null

《医学前沿（英文）》 2017年第11卷第4期页码 502-508 doi: 10.1007/s11684-017-0590-z

摘要：

Although the efficacy of nucleos(t)ide analogue (NA) has been confirmed for treatment of chronic hepatitis B, long-term therapy has been recommended due to the high frequency of off-therapy viral DNA rebound and disease relapse. In this review, the RNA virion-like particles of hepatitis B virus (HBV) are integrated into the life cycle of HBV replication, and the potential significance of serum HBV RNA is systematically described. The production of HBV RNA virion-like particles should not be blocked by NA; in this regard, serum HBV RNA is found to be a suitable surrogate marker for the activity of intrahepatic covalently closed circular DNA (cccDNA), particularly among patients receiving NA therapy. Therefore, the concept of virological response is redefined as persistent loss of serum HBV DNA and HBV RNA. In contrast to hepatitis B surface antigen (HBsAg) that can originate from either the cccDNA or the integrated HBV DNA fragment, serum HBV RNA, with pregenomic RNA origination, can only be transcribed from cccDNA. Therefore, the loss of serum HBV RNA would likely be a promising predicator for safe drug discontinuation. The clinical status of consistent loss of serum HBV RNA accompanied with low serum HBsAg levels might be implicated as a “para-functional cure,” a status nearly close to the functional cure of chronic hepatitis B, to distinguish the “functional cure” characterized as serum HBsAg loss with or without anti-HBs seroconversion.

关键词： chronic hepatitis B serum HBV RNA nucleos(t)ide analogs virological response para-functional cure

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immune escape and microenvironment between RG-like and pri-OPC-like glioma revealed by single-cell RNA-seq

《医学前沿（英文）》 doi: 10.1007/s11684-023-1017-7

摘要： The association of neurogenesis and gliogenesis with glioma remains unclear. By conducting single-cell RNA-seq analyses on 26 gliomas, we reported their classification into primitive oligodendrocyte precursor cell (pri-OPC)-like and radial glia (RG)-like tumors and validated it in a public cohort and TCGA glioma. The RG-like tumors exhibited wild-type isocitrate dehydrogenase and tended to carry EGFR mutations, and the pri-OPC-like ones were prone to carrying TP53 mutations. Tumor subclones only in pri-OPC-like tumors showed substantially down-regulated MHC-I genes, suggesting their distinct immune evasion programs. Furthermore, the two subgroups appeared to extensively modulate glioma-infiltrating lymphocytes in distinct manners. Some specific genes not expressed in normal immune cells were found in glioma-infiltrating lymphocytes. For example, glial/glioma stem cell markers OLIG1/PTPRZ1 and B cell-specific receptors IGLC2/IGKC were expressed in pri-OPC-like and RG-like glioma-infiltrating lymphocytes, respectively. Their expression was positively correlated with those of immune checkpoint genes (e.g., LGALS3) and poor survivals as validated by the increased expression of LGALS3 upon IGKC overexpression in Jurkat cells. This finding indicated a potential inhibitory role in tumor-infiltrating lymphocytes and could provide a new way of cancer immune evasion.

关键词： single-cell RNA-seq glioma radial glia primitive oligodendrocyte precursor cell immune escape

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Construction and identification of lentiviral RNA interference vector of rat leptin receptor gene

Zhengjuan LIU, Jie BIAN, Yuchuan WANG, Yongli ZHAO, Dong YAN, Xiaoxia WANG

《医学前沿（英文）》 2009年第3卷第1期页码 57-60 doi: 10.1007/s11684-009-0003-z

摘要： Leptin resistance is a main mechanism of acquired childhood obesity, and the suppression of long form of leptin receptor (OBRb) gene expression in diet-induced obese rats indicates that the down-regulation of OBRb gene expression plays a pivotal role in the mechanism of leptin resistance. The aim of the present study was to construct the lentiviral RNA interference (RNAi) vector of rat OBRb gene and evaluate the effects of siRNA on silencing OBRb gene expression. The target sequence of siRNA-OBRb was designed, and the complementary DNA containing both sense and antisense oligonucleotides was synthesized. After phosphorylation and annealing, these double-stranded DNA was cloned to pRNA-lentivector-VGFP to construct pRNA-Lenti-OBRb-VGFP recombinants with U6-containing promoter, target sequence and Poly III terminator. Then, the products were confirmed by electrophoresis and sequencing analysis, and the effects of RNAi on reducing gene expression were further confirmed by real-time polymerase chain reaction in transfected rat glioma cells expressing OBRb. The target sequence of siRNA-OBRb was successfully cloned to pRNA-lentivector-VGFP, and the RNAi protocol specifically reduced the expression of OBRb mRNA by approximately 80% compared with controls in transfected rat glioma cells. The successful construction of rat lentivirus vectors expressing OBRb-specific shRNA may be useful for further investigation .

关键词： receptors leptin RNA interference lentivirus vector