IGF-1R硫代反义寡核苷酸抗人肝癌裸鼠原位移植瘤的研究

目的探讨IGF-1R硫代反义寡核苷酸（IGF-1R APSODN）对人肝癌裸鼠原位移植瘤的治疗效果及毒性作用。方法构建40只人肝癌裸鼠原位移植瘤模型，设溶剂对照组、IGF-1R APSODN（75，50，25mg／kg）三剂量组及氟尿嘧啶阳性对照组，各组裸鼠静脉给药25次，用药期间监测裸鼠体重变化，实验结束后行全血分析，肝肿瘤体积、重量测量，计算抑瘤率，病理组织学检查。重复实验一次。结果两次实验IGF-1R APSODN各剂量组肝肿瘤体积和重量均小于溶剂对照组，其中75mg／kg组抑瘤率分别为76．36％、71．81％，50mg／kg组抑瘤率分别为55．65％、61．74％，25mg／kg组抑瘤率分别为47．72％、50．34％。结论IGF-1R APSODN是一种高效、低毒的抗肝癌生物学药物。     

腺病毒介导反义细胞外信号调节蛋白激酶2基因转染可延缓移植肾慢性纤维化进程

严重制约肾移植受者长期存活的重要因素是慢性移植。肾纤维化。慢性移植肾纤维化由免疫学和非免疫学因素引起,临床上一般通过调整免疫抑制方案和其他辅助方法予以治疗,目前还没有直接对移植肾纤维化起作用的药物或治疗方案。我们的研究试图寻找可以保护移植物,阻止或延缓移植肾纤维化的方法和相应的治疗靶点。     

脂质体介导的端粒酶反义寡核苷酸体外转染率及对人膀胱癌EJ细胞的影响

目的通过以脂质体为载体介导端粒酶反义寡核苷酸(antisense oligodeoxynucleotide,asODN)转染人膀胱癌EJ细胞,促进反义寡核苷酸对膀胱癌EJ细胞的生长抑制。方法利用脂质体为载体将针对端粒酶RNA模板区的asODN转染膀胱癌EJ细胞;荧光显微镜观察细胞转染率;PCR-ELISA法测定端粒酶活性;MTT法检测反义寡核苷酸对膀胱癌细胞的抑制。结果脂质体介导的asODN在膀胱癌EJ细胞中的转染率1/2h、4h、8h分别为15%、56%、80%,并且细胞的端粒酶活性和细胞生长明显被抑制,与对照组比较,具有显著性差异(p(0.05)。结论脂质体介导的端粒酶asODN能够有效抑制膀胱癌EJ细胞端粒酶活性和生长,可应用于实验性肿瘤基因治疗研究和端粒酶与膀胱癌关系的进一步研究。     

腺病毒载体介导反义Src基因治疗胶质瘤

目的 探讨腺病毒载体介导的反义Src基因对胶质瘤生长的作用及其机制.方法 应用携带反义Src基因的重组腺病毒体外感染C6胶质瘤细胞,观察其对细胞形态、生长曲线和克隆形成率的影响.建立大鼠胶质瘤动物模型,原位注射重组腺病毒.观察其治疗作用,Western blot检测肿瘤组织Src、Ras、MAPK蛋白的表达,实时逆转录-聚合酶链反应(Real-time RT-PCR)检测肿瘤组织Caspase-3、Caspase-8 mRNA的表达.结果 体外研究表明感染反义Src基因的C6胶质瘤细胞生长受到显著抑制.体内研究表明应用携带反义Src基因的腺病毒原位注射治疗大鼠胶质瘤能够抑制肿瘤生长,减少Src、Ras、MAPK蛋白的表达,增加Caspase-3、Caspase-8 mRNA的表达.结论 腺病毒载体介导的反义Src基因能够抑制大鼠胶质瘤细胞的生长.Src-Ras-MAPK信号通路参与胶质瘤的生长,抑制该信号通路可以增加凋亡通路关键蛋白Caspase-3、Caspase-8的表达.     

Genetic variants within ANRIL (antisense non coding RNA in the INK4 locus) are associated with risk of psoriasis

BackgroundPsoriasis is a systemic inflammatory disease which mostly affects skin. Evidences support the role of autoimmune responses in this disorder. The long non-coding RNA (lncRNA) antisense non coding RNA in the INK4 locus (ANRIL) has been shown to participate in modulation of immune response and in the pathogenesis of immune-related disorders.MethodsWe genotyped four single nucleotide polymorphisms (SNPs) with this lncRNA (rs1333045, rs1333048, rs4977574 and rs10757278) in 286 patients with psoriasis and 300 age-/sex-matched controls to identify the role of ANRIL as a risk locus for psoriasis.ResultsThe C allele of rs1333048 SNP was significantly more prevalent among cases compared with controls (OR (95% CI) = 1.56 (1.23–1.97), adjusted P value = 8.31E−4). The A allele of the rs4977574 had a protective effect against psoriasis (OR (95% CI) = 0.63 (0.49–0.81), adjusted P value = 0.001). The G allele of the rs10757278 conferred risk of psoriasis in the assessed population (OR (95% CI) = 1.9 (1.51–2.4), adjusted P value = 2.18 E−7). The C A G A haplotype (rs1333045, rs1333048, rs4977574 and rs10757278, respectively) was reported to be a protective haplotype against psoriasis (OR (95% CI) = 0.5 (0.35–0.71), adjusted P value = 0.001). The C A G G and T C G G haplotypes conferred risk of psoriasis in the assessed population (OR (95% CI) = 2.37 (1.59–3.54), adjusted P value = 2.4E−4; OR (95% CI) = 5.42 (2.88–10.22), adjusted P value = 1.1E−7, respectively).ConclusionConsequently, ANRIL can be regarded as a risk locus of psoriasis in the assessed population. Future studies are needed to verify whether this contribution is exerted through modulation of immune responses.     

A segment of cold shock protein directs the folding of a combinatorial protein

It has been suggested that protein domains evolved by the non-homologous recombination of building blocks of subdomain size. In earlier work we attempted to recapitulate domain evolution in vitro. We took a polypeptide segment comprising three beta-strands in the monomeric, five-stranded beta-barrel cold shock protein (CspA) of Escherichia coli as a building block. This segment corresponds to a complete exon in homologous eukaryotic proteins and includes residues that nucleate folding in CspA. We recombined this segment at random with fragments of natural proteins and succeeded in generating a range of folded chimaeric proteins. We now present the crystal structure of one such combinatorial protein, 1b11, a 103-residue polypeptide that includes segments from CspA and the S1 domain of the 30S ribosomal subunit of E. coli. The structure reveals a segment-swapped, six-stranded beta-barrel of unique architecture that assembles to a tetramer. Surprisingly, the CspA segment retains its structural identity in 1b11, recapitulating its original fold and deforming the structure of the S1 segment as necessary to complete a barrel. Our work provides structural evidence that (i) random shuffling of nonhomologous polypeptide segments can lead to folded proteins and unique architectures, (ii) many structural features of the segments are retained, and (iii) some segments can act as templates around which the rest of the protein folds.     

Field effect in oral precancer as assessed by DNA flow cytometry and array-CGH

J Oral Pathol Med (2012) 41 : 119–123 Objective: ‘Field cancerization’ is an accepted model for oral carcinogenesis. So far, genetically altered fields have been just reported in the presence of carcinomas. This study assessed the distant mirror fields (MFs) of oral precancer by DNA high‐resolution flow cytometry (hr DNA‐FCM) and array‐Comparative Genomic Hybridization (a‐CGH). Methods: Five leukoplakias without dysplasia (OLs), seven dysplastic leukoplakias (DOLs), and 12 corresponding visually normal and non‐dysplastic MFs were analyzed. DNA aneuploidy (DNA Index, DI ≠ 1) was detected by hr DNA‐FCM on DAPI stained nuclei suspensions. The epithelial DNA aneuploid subclones were FCM‐sorted to obtain genomic DNA for a‐CGH. Results: Mirror fields, OLs, and DOLs showed increasing prevalence of DNA aneuploidy of, respectively, 8%, 20%, and 57%. The average number of chromosome aberrations (Ch‐Abs) was 2.8 in MFs, 3 in OLs, and 10.6 in DOLs. MFs relative to OLs and DOLs had average numbers of Ch‐Abs, respectively, of 1.8 and 3.6. Ch‐Abs were also observed in DNA diploid sublines, and often the same aberrations were observed in both MFs and corresponding OLs/DOLs. Conclusion: DNA aneuploidy and Ch‐Abs in MFs, the last ones being mainly gains, indicate an early onset of field effect in oral carcinogenesis.     

New routes in frontotemporal dementia drug discovery

Introduction: Research into the pathogenic mechanisms behind frontotemporal dementia (FTD) has yielded several new targets for therapeutic intervention; such targets include specific new pathways uncovered by mutations as well as targets involving the modulation, formation and degradation of protein aggregates.

Areas covered: Herein, the authors outline the principal molecular causes underlying FTD to date and the research that has been performed in these areas with respect to an eventual corrective strategy.

Expert opinion: While it is worthwhile targeting pathways affected by specific mutations with a causative loss of function linked to FTD, research still has to contend with issues including the remaining presence of protein aggregates or that treatments are rarely universally applicable. Aiming to recover function in a downstream target caused by the protein aggregates will likely be insufficient due to the large cascade of events affected. It is our belief that the clearance of these aggregates and the inhibition of protein misfolding are more appropriate and direct routes to an eventual therapy.     

Pharmacological treatment options for severe hypertriglyceridemia and familial chylomicronemia syndrome

Introduction: A spectrum of disorders, ranging from rare severe cases of homozygous null lipoprotein lipase deficiency (LPLD)–familial chylomicronemia syndrome (FCS) to heterozygous missense LPLD or polygenic causes, result in hypertriglyceridemia and pancreatitis. The effects of mutations are exacerbated by environmental factors such as diet, pregnancy, and insulin resistance.

Areas covered: In this review, authors discuss chronic treatment of FCS by ultra-low fat diets allied with the use of fibrates, omega-3 fatty acids, niacin, statins, and insulin-sensitizing therapies depending on the extent of residual lipoprotein lipase (LPL) activity; novel therapies in development target triglyceride (TG)-rich lipoprotein particle clearance. Previously, a gene therapy approach to LPL-alipogene tiparvovec showed that direct targeting of LPL function reduced pancreatitis events. An antisense oligonucleotide to apolipoprotein-C3, volanesorsen has been shown to decrease TGs by 70–80% and possibly to reduce rates of pancreatitis admissions. Studies are underway to validate its long-term efficacy and safety. Other approaches investigating the role of LPL modulating proteins such as angiopoietin-like petide-3 (ANGPTL3) are under consideration.

Expert opinion: Current therapeutic options are not sufficient for management of many cases of FCS. The availability of antisense anti-apoC3 therapies and, in the future, ANGPTL3 therapies may remedy this.