Study On Apoptosis Of A549 Cells Induced By A Novel Thiazolidine Compound And Its Mechanism

Background and objectiveLung cancer, a kind of malignant tumor, is threatening humans with high morbidity and mortality. Many cancer patients are suffering from lung cancer. At present, the chemotherapy is the most important treatment of lung cancer, but the majority of chemotherapeutic drugs are easy to cause toxicity side effects, and many drugs are easy to cause drug resistance in the later period of the treament. So it is very important to look for chemotherapeutic drugs which are of high efficiency, less side effects and can overcome multi-drug resistance effectively.Recently, it is found that thiazolidine compounds have a broad spectrum of biological activities. And thiazolidine compounds are widely used in many other fields, such as agriculture and chemistry. And the thiazolidine compounds can induce differentiation and apoptosis in tumor cells, so they can be used in cancer therapy areas. Based on the anti-tumor activity of thiazolidine compounds, we choose YFZ-38 as the study drug to detect the mechanisms of cultured human lung adenocarcinoma A549 cells, expecting to provide certain theoretical basis for the treatment of lung cancer.MethodsThe transplantation tumor model of lung cancer in nude mice was established to study the inhibitory effect of YFZ-38 on the growth of tumor in vivo after dosing. To investigate the inhibitory effects of YFZ-38 to A549 cells, A549 cells were exposed to different concentrations of YFZ-38. We choose XTT assay to detect the cell viability, and we analysis the suppression effects of YFZ-38 in vitro A549 cells growth. Using inverted microscope to observe the morphology of A549 cells, using fluorescence microscopy to observe the nucleus fragmentation,using giemsa staining assay to observe the morphology changes of apoptotic cell, and using nucleosome ELISA assay kits to detect the rates of apoptosis, we analysis the role of YFZ-38 in inducing A549 cells apoptosis. We choose caspase enzyme activity assay kits to detect changes of enzyme activity in caspase, and choose ELISA assay to detect the role of caspase inhibitors for the apoptosis induced by YFZ-38. The detection of Fas, mFasL and sFasL was to analysis the pathways of apoptosis.ResultsThe results of YFZ-38 on the proliferation of A549 cells in vivo showed that YFZ-38 could inhibit the growth of tumors in nude mice in a time-dependent manner. The results of XTT assay showed that YFZ-38 could inhibit the growth of A549 cells in vitro in a dose-dependent manner.And YFZ-38 showed less toxicity to normal cells. The results of A549 cells apoptosis induced by YFZ-38 showed that A549 cells treated with 2.8μM YFZ-38 for 48 h began to show the morphological changes of apoptosis. The results of Giemsa Staining and Hoechst33258 staining also showed the morphological changes of apoptosis, such as dges wrinkled, volume reduced,and apoptotic bodies. The results of nucleosome ELISA kits for the detection of nucleosome levels showed that A549 cells treated with 2.8μM and 5.6μM YFZ-38 for 6h, 12 h, 24 h, 48 h began to show the increased nucleosome levels. The results indicate that YFZ-38 can induce apoptosis of A549 cells in a time-dependent and dose-dependent manner. The results of apoptotic mechanism of A549 cells induced by YFZ-38 showed that YFZ-38 could kill A549 cells in apoptotic way by endoplasmic reticulum, mitochondrial and the Fas involved in.Conclusions1. YFZ-38 can inhibit the multiplication of A549 cells in vitro and in vivo;2. YFZ-38 can induce apoptosis of A549 cells;3. The apoptosis in A549 cells induced by YFZ-38 is based on the endoplasmic reticulum,mitochondrial, and the Fas.