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Single-stranded RNA as primers of terminal deoxynucleotidyl transferase for template-independent DNA polymerization

Chinese Chemical Letters 2023年第2期34卷 161-164页

作者： Houyu Han Jingyu Cui Dianming Zhou Deping Hua Weipan Peng Mengyao Lin Yating Zhang Fangya Li Xiaoqun Gong Jianyu Zhang School of Life Sciences Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology(Tianjin)Tianjin 300072China School of Pharmaceutical Science and Technology Tianjin UniversityTianjin 300072China Tianjin Key Laboratory of Pathogenic Microbiology of Infectious Disease Tianjin Centers for Disease Control and PreventionTianjin 300011China Department of Toxicology Tianjin Centers for Disease Control and PreventionTianjin 300011China

terminal deoxynucleotidyl transferase(Td T) has been characterized as template-independent polymerase using single-stranded DNA(ss DNA) as primers to generate random oligonucleotides. However, the extension performance of Td T to single-stranded RNA(ss RNA) is vague. By systematically comparing and contrasting the performance of Td T-catalyzed ss DNA and ss RNA extension, it is indicated that the catalytic efficiency of ss RNA as primers was about 3 times lower than ss DNA as primers. Collectively, it is believed that understanding the catalytic performance of Td T will help to design the strategy to synthesize chimeric DNA on 3-OH of ss RNA, which becomes invaluable.

关键词： terminal deoxynucleotidyl transferase Single-stranded RNA Single-stranded DNA Template-independent Polymerization

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Transformation of marginal zone lymphoma into high-grade B-cell lymphoma expressing terminal deoxynucleotidyl transferase:A case report

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World Journal of Clinical Cases 2024年第15期12卷 2655-2663页

作者： Zhi-Min Fan Dao-Lei Wu Neng-Wen Xu Li Ye Li-Ping Yan Lin-Jie Li Jun-Yu Zhang Department of Hematology Lishui Municipal Central HospitalLishui 323000Zhejiang ProvinceChina Department of Ophthalmology Lishui Municipal Central HospitalLishui 323000Zhejiang ProvinceChina Department of General Practice Lishui Municipal Central HospitalLishui 323000Zhejiang ProvinceChina Department of Pathology Lishui Municipal Central HospitalLishui 323000Zhejiang ProvinceChina

BACKGROUND High-grade B-cell lymphoma(HGBL)is an unusual malignancy that includes myelocytomatosis viral oncogene(MYC),B-cell lymphoma-2(BCL-2),and/or BCL-6 rearrangements,termed double-hit or triple-hit lymphomas,and HGBL-not otherwise specific(HGBL-NOS),which are morphologically characteristic of HGBL but lack MYC,BCL-2,or BCL-6 *** is partially transformed by follicular lymphoma and other indolent lymphoma,with few cases of marginal zone lymphoma(MZL)*** often has a poor prognosis and intensive therapy is currently mainly advocated,but there is no good treatment for these patients who cannot tolerate *** SUMMARY We reported a case of MZL transformed into HGBL-NOS with TP53 mutation and terminal deoxynucleotidyl transferase *** analysis revealed the gene expression profile was identical in the pre-and post-transformed tissues,suggesting that the two diseases are homologous,not secondary *** chemotherapy was ineffective and the side effect was severe,so we tried combination therapy including venetoclax and *** patient tolerated treatment well,and reached partial *** patient had recurrence of hepatocellular carcinoma and died of multifunctional organ *** survived for 12 months after *** Venetoclax combined with obinutuzumab might improve the survival in some HGBL patients,who are unsuitable for chemotherapy.

关键词： Marginal zone lymphoma High-grade B-cell lymphoma terminal deoxynucleotidyl transferase Venetoclax TP53 mutation Case report

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The Possible Involvement of Apoptotic Decay of terminal deoxynucleotidyl transferase-Positive Lymphocytes in the Reutilization of the Extracellular DNA Fragments by Surrounding Living Cells

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Open Journal of Biophysics 2021年第4期11卷 371-382页

作者： Aleksei N. Shoutko Department of Fundamental Research Russian Scientific Center of Radiology and Surgical Technologies Ministry of Health Care RF Saint-Petersburg Russia

The migrating TdT+ thymocytes can die in other tissues, promoting the surrounding cells’ renewing likes holocrine secretion does. To clarify the role of TdT-enzyme for this function of progenitor lymphocytes, their extracellular media with its components included by living cells analyzed in vitro before and after in vivo irradiation of donor rats. The nucleoid with DNase-sensitive (free) DNA and TdT activity discovered in extracellular media conditioned preliminary by spontaneous apoptotic death of a minor part of the thymocyte’s suspension in vitro. The penetration of labeled products of non-template synthesis with free DNA’ primers from media into cells by pinocytosis confirmed by exogenous polymeric DNA marked artificially. The DNA penetration into cells follows an increase of the cell’s viability and acceleration of spontaneous intracellular DNA-synthesis controlled with labeled thymidine uptake. Both phenomena are typical for either the lowest initial concentration of intact cells or their preliminary irradiation in vivo. The data point to possible involvement of apoptotic decay of TdT+ cells in the reutilization of the extracellular DNA fragments for reparation/regeneration of surrounding living cells.

关键词： Thymocytes Apoptosis terminal deoxynucleotidyl transferase Extracellular Activity DNA Synthesis Precursors Reutilization

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STUDY ON ANTITUMOR DRUG-INDUCED APOPTOSIS IN HUMAN CANCER CELLS BY terminal DEOXYNU-CLEOTIDYL transferase ASSAY

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Chinese Journal of Cancer Research 1997年第3期9卷 3-7页

作者：童彤孙含笑刘丽影梁金凤郭素萍韩乃君程书钧

Objective: Considerable evidence has showed that apoptosis is involved in both cancer development and inhibition. A new assay (terminal deoxynucleotidyl tansferase, TdT) was recently reported to have advan tages in the detection of apoptosis. In this study, this assay was used to investigate antitumor drug induced apoptosis in human cancer cells. Methods: TdT assay, DNA gel electrophoresis, electron and light microscopy were used to observe apoptosis. Results: Our results showed that cisplatin induced apoptosis in both HL 60 and SV40T transformed human bronchial epithelial cells was detected with a good dosage and time response. The occurrence of the apoptosis was preceded by the decrease of bcl 2 mRNA expression. With the TdT assay, apoptotic cells were observed in ovarian tumor of patients treated with carboplatin. Conclusion: TdT assay may be applicable to monitor apoptosis in human cancers induced by chemotherapy, and to evaluate tumor cell response during treatment.

关键词： Apoptosis terminal deoxynucleotidyl transferase Ovarian neoplasms Cisplatin Oncogenes.

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Ultrasensitive Electrochemical Aptasensor Based on Surface-Initiated Enzymatic Polymerization

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Chinese Journal of Chemistry 2016年第3期34卷 337-341页

作者： Pengjuan Wang Ying Wan Yan Su Shengyuan Deng Shulin Yang Sehool of Mechanical Engineering Nanjing University of Science and Technology Nanjing Jiangsu 210094 China School of Environmental and Biological Engineering Nanjing University of Science and Technology Nanjing Jiangsu 210094 China

Coupled with the ability of fast and quantitative response, electrochemical aptasensors （EA） have great potential in the application of early diagnosis of cancer biomarker. In order to determine the rare hiomarkers in the compli- cate clinical samples, scientists are making unremitting efforts towards improving the sensitivity and selectivity of EA. Herein, a ＂sandwich＂-strucmre electrochemical aptasensor （SEA） is developed for analysis of cancer biomarker carcino-embryonic antigen （CEA）. Two DNA aptamers are employed, one of which is thiolated at Y-terminal and immobilized onto the gold electrode as a capture probe, while the other one is served as signal probe. The two ap- tamers could ＂sandwich＂ the target and signal probe is then subjected to the terminal deoxynucleotidyl transferase （TdT）-catalyzed incorporation of biotin labeled dNTPs into its 3＇-terminal. Thus the as-generated long DNA oligo tails allow specific binding of numerous avidin modified horseradish peroxidase （Av-HRP）, resulting in enhanced peroxidase catalyzed electrochemical signals. This signal amplification strategy is termed as surface initiated enzy- matic polymerization （SIEP）. This SIEP amplified SEA has a detection limit of 10 pg·mL^-1, indicating the out- standing amplification efficiency.

关键词： electrochemical aptasensor terminal deoxynucleotidyl transferase surface initiated enzymaticpolymerization "sandwich"-structure sensitive

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