背景与目的:靶向药物治疗(targeted drug therapy)逐渐成为是目前肿瘤治疗的重要手段。近年来的研究发现,磷脂酰丝氨酸(Phosphatidylserine,PS)是抗肿瘤治疗的重要靶点之-。膜联蛋白B1(annexin B1)是孙树汉教授克隆的一个新的膜联蛋白(A...
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背景与目的:靶向药物治疗(targeted drug therapy)逐渐成为是目前肿瘤治疗的重要手段。近年来的研究发现,磷脂酰丝氨酸(Phosphatidylserine,PS)是抗肿瘤治疗的重要靶点之-。膜联蛋白B1(annexin B1)是孙树汉教授克隆的一个新的膜联蛋白(Annexins)亚家族成员,对PS的结合具有很高的亲和力和特异性。蜂毒肽(melittin,MLT)是蜂毒的主要成分,具有很强的抗肿瘤作用。以肿瘤内皮细胞暴露的磷脂酰丝氨酸为"靶",以AnxB1为"导向分子",以蜂毒素为"抗肿瘤分子",构建基因工程靶向抗肿瘤药物mAnxB1-MLT。方法:利用PCR重叠区延伸法连接AnxB1-MLT,融合基因克隆至pGEX5T融合表达载体和pET-28a表达载体,转染大肠杆菌,分别就IPTG浓度、利福平、表达温度等条件进行优化。结果:融合蛋白AnxB1-MLT在pET-28a表达载体中无明显表达条带。37℃条件下,pGEX-5T表达质粒中高效表达,表达产物占菌体总蛋白的40%以上;IPTG、利福平对表达影响较小,但表达产物均为包涵体,不利于纯化。在22-25℃条件下诱导表达,虽然大肠杆菌生长较慢,但表达产物均为可溶性表达,且占菌体蛋白30%以上。利用GST亲和层析柱纯化,蛋白纯度大于90%。结论:成功构建了融合表达蛋白AnxB1-MLT的表达质粒,优化了诱导表达条件,获得了重组蛋白,为进一步通过动物模型研究其靶向抗肿瘤活性奠定了基础。
Background & Aims:H19 is one of the earliest identified and studied long noncoding *** is presumed that H19 is essential for regulating development and disease conditions,and is associated with ca
Background & Aims:H19 is one of the earliest identified and studied long noncoding *** is presumed that H19 is essential for regulating development and disease conditions,and is associated with ca
Background & Aims:The liver is frequently subject to insult because of viral infection,alcohol abuse,or toxic chemical *** research has been conducted to identify blood markers that can better discern liver damage...
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Background & Aims:The liver is frequently subject to insult because of viral infection,alcohol abuse,or toxic chemical *** research has been conducted to identify blood markers that can better discern liver damage,but little progress has been achieved in clinical ***,circulating microRNAs(miRNAs) have been reported as potential biomarkers for the noninvasive diagnosis of *** & Results:In this study,we investigated whether plasma miRNAs have diagnostic utility in identifying liver *** study was divided into 2 phases:marker selection by real-time quantitative PCR analysis of a small set of plasma samples,and marker validation with a large set of plasma samples from patients with chronic hepatitis B viral infections and healthy *** mouse model systems, D-galactosamine- and alcoholinduced liver injury,were also developed to evaluate whether differences in miRNA concentration were associated with various liver *** the miRNA candidates identified,miR-122 presented a disease severity-dependent change in plasma concentration in the patients and animal *** with an increase in aminotransferase activity in the blood,the change in miR-122 concentration appeared ***,this change was more specific for liver injury than for other organ damage and was more reliable, because the change was correlated with liver histologic ***:Our findings suggest that circulating miR-122 has potential as a novel,predictive,and reliable blood marker for viral-, alcohol-,and chemicalinduced liver injury.
Background & Aims:Although numerous long non-coding RNAs(IncRNAs) have been identified in mammals,many of their biological roles remain to be ***, long non-coding RNAs(LncRNAs) encoded in human genome have been sh...
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Background & Aims:Although numerous long non-coding RNAs(IncRNAs) have been identified in mammals,many of their biological roles remain to be ***, long non-coding RNAs(LncRNAs) encoded in human genome have been shown to serve as important regulatory transcripts and may have other previously unappreciated *** can serve as scaffolds,aiding the association of proteins with genomic *** our recent study,we identified nonoverlapping signatures of a small number of IncRNAs that are aberrantly expressed in human Hepatitis B virus(HBV)-related HCC compared with matched peritumoral tissues. Among these IncRNAs,H19 was found to play a potential role in HCC *** reports suggest that H19 contributes to carcinogenesis,including hepatocellular carcinoma(HCC). Examination of the Oncomine resource showed that most HCC cases express H19 at a level that is comparable with the liver,with a tendency toward lower *** is consistent with our previous microarray data and indicates a more complicated role of H19 in HCC that needs to be *** & Results:In this study,the expression level of H19 was assessed in different regions of HCC patients'liver ***,our results showed that the H19 RNA was enriched in the sections around the tumor *** is in agreement with a previousreport that H19 was most prominent at the boundary of the tumor *** collected tissues from different sites of the patients'livers,and we defined the peritumoral tissues as the sections that were 2 cm away from the tumor tissues and the normal tissues as the sections that were at least 3 cm away from the ***- and gain-of-function studies on this IncRNA in the HCC cell lines,SMMC7721 and HCCLM3,were used to characterize its effects on gene expression and to assess its effect on HCC metastasis both in vitro and in *** this study,we show that HI9 was underexpressed in intratumoral HCC tissues(T),as compared with peritumora
Background & Aims:The involvement of the hepatitis B virus X(HBx) protein in epigenetic modifications during hepatocarcinogenesis has been previously *** noncoding RNAs(IncRNAs),a kind
Background & Aims:The involvement of the hepatitis B virus X(HBx) protein in epigenetic modifications during hepatocarcinogenesis has been previously *** noncoding RNAs(IncRNAs),a kind
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