Study Of DNA Microarray On Apoptosis Of Human Hepatocarcinoma Cells Induced By Arsenic Trioxide In Vitro

Objective: It has been proved that apoptosis (APO) obeys some laws and orders. Many factors, such as physi-chemical and biological factors, drugs and traditional herbs can induce the APO of many kinds of tumor cells. It has been becoming the hot spot in the fields of tumor treatment to induce the APO of tumor cells. In order to understand how arsenic trioxide induced the APO of human hepatocarcinoma cell, BEL-7402 , on the molecular level, the gene chip or DNA microarray had been used to detect the expression panel of the genes associated with apoptosis of BEL-7402 cells treated by arsenic trioxide.Methods: BEL-7402 cells had been used to testify the APO induced by arsenic trioxide with the methods of MTT, agarose gel electrophoresis, TUNEL, transmission electron microscopy and FCM techniques. Then, the apoptotic BEL-7402 cells were tested with the method of DNA microarray to obtained the expression panel of the genes associated with apoptosis. Results: The experiment showed that arsenic trioxide had apparent inhibition on BEL-7402 cells in vitro. The inhibition resulted from the APO induced by arsenic trioxide. Under certain conditions, the concentration of arsenic trioxide inducing the APO on 50% of BEL-7402 cells was 2umoI/L. Typical apoptotic changes, such as apoptotic DNA strand, apoptotic cellpeak and apoptotic body etc, had been observed with the methods of agarose gel electrophoresis, FCM techniques and transmission electron microscopy. In the DNA microarray test, there were five pieces of genes concerned with APO expressed divergently. Those were HIP2, UBE2D3, Apaf-1, BCL10 and TRAF-3. Except HIP2, all other genes had a higher expression. Conclusion: Combining recent research on APO with the results of the experiment, fallowings can be concluded: 1, Typical apoptotic changes of human hepatocarcinoma BEL-7402 cells can be induced by arsenic trioxide under certain conditions in vitro. 2, Five pieces of genes, HIP2, UBE2D3, Apaf-1, BCL10 and TRAF-3, associated with APO expressed divergently. 3, Arsenic trioxide as an apoptotic signal induces the APO of BEL-7402 cells by starting both the death receptor route and the mitochondria mediated route, getting caspase3 activating and initiating the cascade of the caspase finally. Above findings could lay foundation for further researching on tumor cell apoptosis and the new method of human hepatocarcinoma treatment.