Arsenic trioxide (AsO) is an old drug that has recently been reintroduced as a therapeutic agent for acute promyelocytic leukemia (APL). Although AsO is also applied to treat other types of cancer in vitro and in vivo, it has been reported that single agent AsO has poor efficacy against non-hematologic malignant cancers in clinical trials. Recently, a few reports have indicated that organic arsenic compounds can be a possible alternative for the treatment of AsO-resistant cancers. In this study, we aimed to investigate whether the organic arsenic compound phenylarsine oxide (PAO) has potent cytotoxic effects against human hepatocellular carcinoma (HCC) HepG2 cells. Our results showed that PAO not only had a potent inhibitory effect on the proliferation of HepG2 cells but also activated apoptosis-related proteins (e.g., caspase-3 and -9 and poly-ADP ribose polymerase) in a dose- and time-dependent manner. Furthermore, intracellular ROS were specifically accumulated in the mitochondria and endoplasmic reticulum (ER) after exposure to PAO, implying that they are the target organelles for PAO-induced cytotoxicity. Additionally, when the cells were pretreated with antioxidant N-acetylcysteine (NAC), apoptosis and ER-stress were attenuated significantly, suggesting that induction of apoptosis and cell death probably occurs through the ROS-mediated mitochondria and ER-stress dependent signaling pathways.
In this study, the new technological parameters have been optimized for running Single factor experiment and orthogonal design test. Furthermore, the optimal extractible condition of SOD activity from Alfalfa was determined by the assay of NBT photoreduction method. The results reveal that when the ratio of liquor to material is 1 to 10; pH value of phosphate buffer solution is 7.5; ultrasonic power is 400 w, therefore, the SOD activity of Alfalfa is up to 66.54 U/mL. In addition, we characterized the activity analysis of Alfalfa proteins using heat denaturation, ammonium sulfate precipitation and ultraviolet spectroscop. The results showed that the protein removal rate is approximate 91.70 % and SOD specific activity is 6.5 times. Based on the above results, we reached a preliminary estimate that the SOD from Alfalfa is a kind of Cu/Zn-SOD.
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