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Tumor gene mutations and messenger rna expression： correlation with clinical response to icotinib hydrochloride in non-small cell lung cancer

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Chinese Medical Journal 2011年第1期124卷 19-25页

作者： REN Guan-jun ZHAO Yuan-yua ZHU Yu-jia XIAO Yi XU Jia-sen SHAN Bin ZHANG Li Department of Respiration Peking Union Medical College Hospital Peking Union Medical College & Chinese Academy of Medical Sciences Beijing 100730 China SurExam Bio-Tech- Co. Ltd. Science City Guangzhou Guangdong 510663 China Gleneagles Clinical Research Centre Beijing 100062 China

Background Molecular targeted drugs is now widely used in non-small cell lung cancer （NSCLC） clinical treatment. Icotinib hydrochloride is a new type of oral epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （EGFR-TKIs）. In this study, we examined the role of EGFR, K-RAS, B-RAF somatic mutations and EGFR mrna expression in tumor specimens from advanced NSCLC patients as predicators of the efficacy of icotinib hydrochloride. Methods We analyzed tumor paraffin-embedded specimens, which were obtained from 14 of 40 patients with advanced NSCLC who enrolled in the stage I clinical trial of icotinib hydrochloride. Somatic mutations were evaluated by mutant-enriched liquidchip （MEL） technology, and EGFR mrna expression was measured by branched DNA liquidchip （MBL） technology. Results In the 14 specimens, seven patients showed EGFR mutations, exon 19 deletion （3/7） and exon 21 point mutation （4/7）; and two patients showed K-RAS mutation. No mutations in EGFR exon 20. or B-RAF were detected. In patients with EGFR mutation, one patient developed progress disease （PD）, three patients had stable disease （SD）, two patients had partial responses （PR） and one patient had a complete response （CR）. In patients with wild-type EGFR, four patients had PD, three patients acquired SD, and none had PR/CR （P=-0.0407）. EGFR mutations were associated with better progress-free survival （PFS）（141 days vs. 61 days） but without a statistically significant difference （P=0.8597）, and median overall survival （OS）（-〉449 days vs. 140 days）. EGFR mrna expression levels were evaluated （three high, eight moderate, one low, and two that can not be measured due to insufficient tumor tissue） and no statistically significant relationships was observed with response, PFS or OS. Conclusions The EGFR mutation rate was consistent with that reported in the Asian population, so the MEL technology is reliable for measuring EGFR mutation with high throughput and rapidity. EGFR exon 19 deletions and exon 21 point mutation are predictive biomark

关键词： non-small cell lung cancer icotinib hydrochloride epidermal growth factor receptor somatic mutation messenger rna

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The messenger rna and Long Non-coding rna Expression Profiles in Ectopic and Eutopic Endometrium Provide Novel Insights into Endometriosis

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Reproductive and Developmental Medicine 2019年第1期3卷 11-17页

作者： Song-Ping Liu Xin Tian Hong-Yan Cui Qiong Zhang Ke-Qin Hua Department of Gynecology Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200090China Department of Obstetrics and Gynecology Zhenjiang Maternal and Child Health HospitalZhenjiang 212001China

Objective:To establish the messenger rna(mrna)and long non-coding rna(lncrna)expression profiles in ectopic and eutopic endometrium and provide novel insights into endometriosis.Methods:The mrna and lncrna expression profiles were tested using high-throughput sequencing technology in ectopic and eutopic endometrium with endometriosis and normal endometrium without endometriosis.The potential targeted lncrnas were annotated by analyzing the correlation between lncrna and mrna expression to better understand the pathogenesis of endometriosis.Results:In ectopic compared with normal endometrium,a total of 2,188 mrnas and 1,200 lncrnas were differentially expressed with a fold-change(FC)≥2.5.In eutopic compared with normal endometrium,a total of 2,324 mrnas and 695 lncrnas were differentially expressed with an FC≥1.5.In ectopic compared with eutopic endometrium,a total of 2,223 mrnas and 511 lncrnas were differentially expressed with an FC≥2.Bioinformatic analysis indicated that the differentially expressed mrnas were enriched in the biological processes and signaling pathways involved in endometriosis.In addition,we constructed a gene coexpression network based on the dysregulated lncrnas in both ectopic endometrium and eutopic endometrium,combined with their coexpressed mrnas to simulate the complex interactions.Conclusions:This study describes the first-to-integrate analysis of the differential expression profiles of mrnas and lncrnas,including analyses between ectopic and normal endometrium,eutopic and normal endometrium,and ectopic and eutopic endometrium,which provides new insights to investigate the pathogenesis of endometriosis and explore novel diagnostic biomarkers and therapeutic targets.

关键词： Endometriosis Gene Profile Long Non-coding rna messenger rna rna Sequencing

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mrna delivery in cancer immunotherapy

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Acta Pharmaceutica Sinica B 2023年第4期13卷 1348-1357页

作者： Yichen Zhong Shi Du Yizhou Dong Division of Pharmaceutics&Pharmacology College of PharmacyThe Ohio State UniversityColumbusOH 43210USA Department of Radiation Oncology Department of Biomedical EngineeringCenter for Clinical and Translational ScienceComprehensive Cancer CenterDorothy M.Davis Heart&Lung Research InstituteCenter for Cancer EngineeringCenter for Cancer MetabolismPelotonia Institute for Immune-OncologyThe Ohio State UniversityColumbusOH 43210USA Icahn Genomics Institute Precision Immunology InstituteDepartment of Oncological SciencesTisch Cancer InstituteFriedman Brain InstituteIcahn School of Medicine at Mount SinaiNew YorkNY 10029USA

messenger rna(mrna)has drawn much attention in the medical field.Through various treatment approaches including protein replacement therapies,gene editing,and cell engineering,mrna is becoming a potential therapeutic strategy for cancers.However,delivery of mrna into targeted organs and cells can be challenging due to the unstable nature of its naked form and the low cellular uptake.Therefore,in addition to mrna modification,efforts have been devoted to developing nanoparticles for mrna delivery.In this review,we introduce four categories of nanoparticle platform systems:lipid,polymer,lipid-polymer hybrid,and protein/peptide-mediated nanoparticles,together with their roles in facilitating mrna-based cancer immunotherapies.We also highlight promising treatment regimens and their clinical translation.

关键词： Cancer immunotherapy Lipid nanoparticles Lipid—polymer hybrid nanoparticles messenger rna mrna delivery Polymeric nanoparticles Protein/peptide-mediated nanoparticles

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Biomedical applications of mrna nanomedicine

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Nano Research 2018年第10期11卷 5281-5309页

作者： Qingqing Xiong Gha Young Lee Jianxun Ding Wenliang Li Jinjun Shi Center for Nanomedicine and Department of Anesthesiology Brigham and Women's Hospital Harvard Medical School Boston MA 02115 USA Department of Hepatobiliary Cancer Tianjin Medical University Cancer Institute & Hospital Tianjin 300060 China School of Pharmacy Jilin Medical University Jilin 132013 China

As an attractive alternative to plasmid DNA, messenger rna （mrna） has recently emerged as a promising class of nucleic acid therapeutics for biomedical applications. Advances in addressing the inherent shortcomings of mrna and in the development of nanoparticle-based delivery systems have prompted the development and clinical translation of mrna-based medicines. In this review, we discuss the chemical modification strategies of mrna to improve its stability, minimize immune responses, and enhance translational efficacy. We also highlight recent progress in nanoparticle-based mrna delivery. Considerable attention is given to the increasingly widespread applications of mrna nanomedicine in the biomedical fields of vaccination, protein-replacement therapy, gene editing, and cellular reprogramming and engineering.

关键词： messenger rna chemical modification nanoparticle vaccination gene editing protein-replacement cellular reprogramming and engineering

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Differential Expression of Microrna in Endothelial Cells Incubated with Serum of Hypertension Patients with Blood-Stasis Syndrome

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Chinese Journal of Integrative Medicine 2015年第11期21卷 817-822页

作者：王铭铭方梅霞陈利国王华强刘红杰唐海兰 Faculty of Traditional Chinese Medicine Medical School of Jinan University

Objective： To screen out blood-stasis syndrome （BSS）-associated microrna and therefore determine the possible target for treating hypertension. Methods： A high-energy sequencing method and digital gene expression sequencing theory were adopted to sequence microrna （mirna） and messenger rna （mrna）, and to determine differential expression in human umbilical vein endothelial cells incubated with serum samples from hypertension patients with or without BSS, and healthy controls. The results were confirmed using gene prediction software. Results： A total of 13 mirnas and 11 mrnas showed statistical difference both in the BSS/normal groups and BSS/non-BSS groups, respectively. Four pairs of target mRNNmirna were identified： FRMD4Nhsa-miR-34a, MAP3K14/hsa-miR-34a, PER1/hsa-miR-34a, and FGF2/hsa-miR-132. Conclusion： Four mRNNmirna pairs mentioned above seem to be involved in pathogenesis and maintenance of hypertension with BSS.

关键词： hypertension blood-stasis syndrome microrna messenger rna Chinese medicine

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Effect of lipopolysaccharide on expression andcharacterization of cholecystokinin receptorsin rat pulmonary interstitial macrophages

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Acta Pharmacologica Sinica 2004年第10期 107-113页

作者： Shun-jiang XU, Wei-juan GAO, Bin CONG, Yu-xia YAO, Zhen-yong GUDepartment of Pathophysiology, Hebei Medical University, Shijiazhuang 050017, ChinaNow in Hebei Province Center for Disease Prevention and Control, Shijiazhuang 050021, China

AIM: To investigate the effect of lipopolysaccharide (LPS) on the expression and the binding characteristics of cholecystokinin receptors (CCK-R) in rat pulmonary interstitial macrophages (PIMs). METHODS: The PIMs isolated from rat lung tissues were purified by the collagenase digestion method combined with alveolar lavage and pulmonary vessel perfusion. The expression of CCK-R mrna was detected by RT-PCR and Southern blot analysis and the binding experiments were performed by radioligand binding assay (RBA). RESULTS: CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR) mrna were detected in rat PIMs and their RT-PCR amplified products had a size of approximately 1.37 kb and 480 bp, respectively. The relative expression of CCK-BR mrna was higher than that of CCK-AR mrna after incubation with LPS for 0.5, 2, and 6 h. The expression of CCK-R mrna could be upregulated obviously by LPS. Southern blot analysis of RT-PCR amplified CCK-AR and CCK-BR mrna products using [γ-32P]ATP 5’-end-labelled probe showed sp

关键词： lipopolysaccharides lung macrophages cholecystokinin receptors messenger rna gene expression radioligand assay

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Downregulation of Kυ4.2 and Kυ4.3 channel gene expression in right ventricular hypertrophy induced by monocrotaline in rat

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Acta Pharmacologica Sinica 2004年第2期 100-104页

作者： Tian-tai ZHANG, Bing CUI, De-zai DAI2 Research Division of Pharmacology, China Pharmaceutical University, Nanjing 210009, China

AIM: To investigate the differences in gene expression of transient outward potassium ion channel between the free wall of right ventricle, free wall of left ventricle, and the septum in monocrotaline (MCT)-induced right ventricular hypertrophy of rat. METHODS: Twenty rats were randomly divided into two groups: a single injection of monocrotaline (MCT) 60 mg/kg (model) or saline (control). Four weeks later, hemodynamic parameters were measured and the gene expression of Ito channels were detected by semi-quantitative RT-PCR. RESULTS: After 28 d, the right ventricular systolic pressure and central venous pressure were remarkably elevated by 128 % and 533 % in the MCT-treated group, accompanied by an overt right ventricle (RV) remodeling. The difference of mrna expression of Kv1.4 was not significant in free wall of RV, left ventricle (LV), and septum in MCT group compared with control group. In contrast, mrna of Kv4.2 and Kv4.3 in the free wall of RV in MCT-induced rat was dramatically decreased by 45.2 % and 51.1 % vs control, however, in free wall of LV and septum, no differ- ence was found. In addition, mrna expression level of Kv4.2 in control rat was significantly lower in septum than that in free wall of RV and LV. CONCLUSION: Expression of Kv1.4, Kv4.2, and Kv4.3 differs between regions in normal rat hearts. The down-regulation of Kv4 family gene expression of Ito contributed to the pathophysiologi- cal changes in ventricular hypertrophy and pulmonary hypertension induced by MCT.

关键词： right ventricular hypertrophy potassium channels messenger rna monocrotaline

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Minimal Secondary Structure Formation on mrnas with a Shine-Dalgarno Sequence for Chromosomal Genes in <i>Rhodobacter sphaeroides</i>

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Advances in Microbiology 2021年第10期11卷 529-540页

作者： Damilola Omotajo Hyuk Cho Madhusudan Choudhary Department of Biological Sciences Sam Houston State University Huntsville Texas USA Cumming School of Medicine University of Calgary Calgary Alberta Canada Department of Computer Science Sam Houston State University Huntsville Texas USA

The Shine-Dalgarno (SD) sequence, when present, is known to promote translation initiation in a bacterial cell. However, the thermodynamic stability of the messenger rna (mrna) through its secondary structures has an inhibitory effect on the efficiency of translation. This poses the question of whether bacterial mrnas with SD have low secondary structure formation or not. About 3500 protein-coding genes in Rhodobacter sphaeroides were analyzed and a sliding window analysis of the last 100 nucleotides of the 5’ UTR and the first 100 nucleotides of ORFs was performed using rnafold, a software for rna secondary structure analysis. It was shown that mrnas with SD are less stable than those without SD for genes located on the primary chromosome, but not for the plasmid encoded genes. Furthermore, mrna stability is similar for genes within each chromosome except those encoded by the accessory chromosome (second chromosome). Results highlight the possible contribution of other factors like replicon-specific nucleotide composition (GC content), codon bias, and protein stability in determining the efficiency of translation initiation in both SD-dependent and SD-independent translation systems.

关键词： Shine-Dalgarno Sequence Secondary Structure messenger rna Translation Initiation

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Establishment of a real-time PCR for the quantification of transforming growth factor beta1 mrna in hepatocellular carcinoma

Establishment of a real-time PCR for the quantification of t...

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中华医学会第七次全国检验医学学术会议

作者： Eastern Hepatobiliary Hospital 200438 Peng Qi

Background:The carcinogenesis of hepatocellular carcinoma (HCC)is a multi-factorial,multistep and complex process.Its prognosis is poor and early detection is of the utmost importance. Transforming growth factorβ1(TGF-β1)message rna(mrna)has been reported to be elevated in HCC patients using Northern blotting.However,little work has been done about the detection of TGF-β1 mrna levels in peripheral blood of patients with HCC using the real-time polymerase chain reactions(PCR)method.The aim of this prospective pilot study was to assess the prognostic value of quantitative levels of TGF-β1 mrna in peripheral blood of patients with HCC.We then investigated whether there was a correlationship between the expression of TGF-β1 mrna in peripheral blood and the many diagnostic and pathological factors. Methods:We developed an optimized Taqman real-time PCR to quantify TGF-β1 mrna in peripheral blood of 53 patients with HCC and 44 healthy volunteers.In addition,blood was collected from patients with HCC for measurement of levels of T.Bil, prealbumin,albumin,ALT,AST,GGT,AFU,AFP,CEA,CA19-9, viral load and platelet counts.Statistical analysis was performed using the SPSS software system(SPSS 10.0).

关键词： Transforming growth factor betal messenger rna Hepatocellular carcinoma Quantitative PCR Tumor size Early diagnosis Molecular marker Peripheral blood.

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