Vol.50,No.9,2023
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Cover Story：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.
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