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The Therapeutic Effects on Hepatocelluar Carcinoma Caused by Eukaryotic Expression of TRAIL
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Abstract:
Objective: To construct an eukaryotic expressing plasmid of mouse TRAIL (mTRAIL), and investigate its apoptosis-inducing ability to hepatocelluar carcinoma cells in vitro and in vivo, inhibitory effect on the growth of hepatocelluar carcinoma, and its synergism with pCH510, an eukaryotic expressing plasmid of recombinant human FN polypeptide. Methods: The eukaryotic expressing plasmid of mTRAIL was constructed by RT-PCR and DNA recombination techniques. Gene transfection was performed in vitro and in vivo. The apoptosis rate of hepatocelluar carcinoma cells was measured by Flow Cytometry. The apoptosis of hepatocelluar carcinoma cells was also detected by TdT-mediated dUTP nick end labeling (TUNEL) and histochemistry techniques. The inhibitory effect of gene transfection on solid tumor was observed in mice. Results: The cDNA of mTRAIL was amplified by RT-PCR from the RNA of the mouse spleen cells, and cloned into the eukaroytic expressing vector pcDNA3.1. The recombinant plasmid was designated as pX1. The BHK cells transfected with plasmid pX1 could attack H22 hepatocelluar carcinoma cells and induce them into apoptosis. The transfection of plasmid pX1 through injection into mouse muscles could inhibit the growth of hepatocelluar carcinoma by inducing tumor cells into apoptosis. Plasmid pX1 and pCH510 have a synergistic inhibitory effect on the hepatocelluar carcinoma growth. Conclusion: Plamid pX1 could be expressed in cells and in vivo in mouse. The expression of pX1 in vivo and in vitro could induce hepatocelluar carcinoma cells into apoptosis and inhibit the growth of hepatocelluar carcinoma by this mechanism. Plasmid pX1 and pCH510 have a synergistic inhibitory effect on the hepatocelluar carcinoma growth.
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