Expression Of Tumor Suppressor Gene ARHI In Hela Cells

Objective: To express Aplysia Ras Homolog I (ARHI) gene and study the biological activity of recombinant ARM protein in HeLa cells. Methods: The ARM gene was got from pcDNA3.0-ARHI by PCR and sequenced. Then recombinant eukaryotic expression plasmid was constructed, identified and expressed in HeLa cells. Further investigation was made to study the expression and the function and biological activity of expression product by means of RT-PCR, MTT, FCM and Hoechst. Results: Electrophoresis in agarose gels indicated that the amplified special product by RT-PCR was about 700 bp as expected. Eukaryotic expression vector was constructed correctly . The green fluorescence was seen after the HeLa cells was transfected for 24 hours. The product showed apoptosis-inducing activity on HeLa cells by means of MTT, FCM and Hoechst. Conclusions: ARM can induce apoptosis of HeLa cells, suggesting that ARM may serve as a new target for gene therapy for the cervix cancer. So this experiment lays the foundation for further research on molecular mechanism on anti-tumors and the gene therapy.