Effect About Cisplatin Combined With Low Molecular Weight Heparin On Human Lung Adenocarcinoma A549 Cells’ Proliferation Inhibition And Expression Of Bcl-2 And Bax

Objective:As a first-generation platinum-based chemotherapy drug, cisplatin is a cycle non-specific drug and can cause termination of DNA and cell death. Research shows that cisplatin anti-tumor is related to the induction of cell apoptosis. Low molecular weight heparin, as an important anticoagulant, has function of anticoagulant, anti-tumor, anti-inflammatory and anti-viral. Recently, the research of inducing apoptosis has drew attention. So far, most experiments are focus only on cisplatin or low molecular weight heparin. We have not found research on their combined effects. Therefore, the objective of this study was to observe the combined effect of cisplatin and low molecular weight heparin on human lung adenocarcinoma A549 cells, and discuss the mechanism of proliferation and apoptosis and to find more effective clinical chemotherapy drugs for improve the life quality of patients suffering from advanced lung cancer. Methods:The human lung cancer A549 cells choosed from logarithmic growth phase, were cultured in HyClone RPMI-1640 medium, incubator maintained 5% CO2 and 37℃. Cells were divided into experimental and control groups. Experimental groups include cisplatin group (2ug/ml DDP), low molecular weight heparin group (10ug/ml LMWH) and the joint group (2ug/ml DDP+10ug/ml LMWH). Control group with no drug. MTT assay was used to detect the absorbance of test groups at 48h or 72h, and then contrast the differences of inhibition rate among the experimental groups and control group. The difference of apoptosis rate of experimental groups and control group were detected by the flow cytometry instrument technology at a 48h treatment. Enzyme-linked immunosorbent assay (ELISA) technique was used to carrying out the different expressions of bcl-2 protein and bax protein among experimental groups and control group. Results:1. The proliferation inhibition rate of A549 cells was stronger in the joint group than DDP group and LMWH group(P<0.05, n=10)and the inhibition is stronger than the 48h treatment(P<0.05,n=10).2. The apoptosis rate of A549 cells was stronger in the joint group than DDP group and LMWH group (P<0.05, n=3).3. The down-regulation expression of bcl-2 protein was stronger in joint group than DDP group and LMWH group(P<0.05, n=10).4. The up-regulation expression of bax protein was stronger in joint group than DDP group and LMWH group(P<0.05, n=10). Conclusion:1. DDP combine LMWH has a stronger effect to restrain proliferation than DDP and LMWH used alone on human lung adenocarcinoma A549 cells in vitro.2. DDP combine LMWH has a stronger effect to induce apoptosis than DDP and LMWH used alone on human lung adenocarcinoma A549 cells in vitro. Therefore, the mechanism of DDP combine LMWH effect proliferation and apoptosis may be connected with down-regulation the expression of bcl-2 protein and up-regulation the expression of bax protein.