1)Beijing Key Laboratory for Radiobiology, Institute of Radiation Medicine, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China;2)Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
This work was supported by grants from The National Natural Science Foundation of China (31570817, 32071290).
Objective To investigate the target RNA cleavage activity of V4-2-Dz a conjugate formed by coupling the aptamer V4-2 of the hepatocellular carcinoma cell-specific membrane protein VASN (vasorin) with the DNAzyme (Dz) targeting the telomerase reverse transcriptase (hTERT), in vitro and in hepatocellular carcinoma cells, and to establish a new strategy for DNAzyme delivery.Methods The secondary structure of V4-2-Dz was predicted by using the Mfold website and the cleavage activity of V4-2-Dz was examined by in vitro cleavage assay. And then, the ability of V4-2-Dz binding to hepatocellular carcinoma HepG2 cells was examined by flow cytometry and laser confocal assay. The effects of V4-2-Dz on the hTERT mRNA expression and cell proliferation of HepG2 cells were analyzed by using RT-qPCR and MTT assays.Results In vitro cleavage assays showed that V4-2-Dz has hTERT RNA cleavage activity. Flow cytometry and laser confocal results showed that V4-2-Dz specifically binds to HepG2 cells and could reduce the hTERT mRNA levels and inhibit cell proliferation significantly.Conclusion The conjugate of aptamer V4-2 with DNAzyme of telomerase reverse transcriptase, V4-2-Dz, has targeted cellular delivery and cleavage activity. The conjugate based on the aptamer and DNazyme provides a new idea for antitumor drug research.
MA Fei-Yue, LI Hui, LIU Xue-Mei, TAI Fu-Min, SHAO Ning-Sheng, GAO Bo, ZHENG Xiao-Fei.Research: Functional Study of Conjugate of VASN Aptamer-DNAzyme for Telomerase Reverse Transcriptase[J]. Progress in Biochemistry and Biophysics,2023,50(9):2175-2184
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