Nolatrexed Hydrochloride In Vitro To Promote The Human Hepatoma Cell Apoptosis In HepG-2 Experimental Study

Objective: Study nolatrexed hydrochloride on cultured human hepatoma cells HepG-2 inhibition of proliferation of hepatoma cells and HepG-2 apoptosis suppressor gene Bcl-2 expression.Methods: MTT colorimetric observation nolatrexed hydrochloride on human hepatoma cell line HepG-2 inhibition of proliferation; by immune- ocytochemical staining of human hepatoma cells HepG-2 apoptosis suppressor gene Bcl-2 expression affected .Results: 1. Before the experiment for the HepG-2 cell adherent of the spindle cells, cells with close cell growth speed, every 2 to 3 days to be mass generation; in hydrochloric acid by adding chemotherapy after 12h nolatrexed cell morphology were observed to round, smaller size, cell density and cell shrinkage, cell membrane and nuclear membrane integrity, mitochondria occurred concentrate and ultra-sexual chromatin condensation, and nuclear Week "collapse" to form one or more massive structure, " dense sphere, "or outside" germination a strand of grape the formation of small spherical samples. Observed after 24 endoscopic vision blurred, the membrane swelling﹑cracking﹑damage.2. Tumor cells in vitro drug susceptibility testing that is, MTT, to experiment with three groups of drugs that (ADM group, DDP group, hydrochloric acid nolatrexed group) to set up four in each group of drug concentration gradient, while the establishment of the control group, results showed that hydrochloric acid nolatrexed of human liver cancer cell growth of HepG-2 significantly inhibited to promote apoptosis of tumor cells, high concentration of growth inhibition rate of 80.15%, with a good dose-effect relationship, in which a high concentration to inhibition rate of the drug group, followed by concentration group for each experimental tumor lines are repeated 3 times, the repeatability of results is good; when the group reaches the maximum when the drug concentration nolatrexed hydrochloride group and positive control Group (ADM group and DDP group) compared to the inhibitory rate of tumor cells is divided into 80.15 percent, 48.12 percent, 51.69 percent, the difference was statistically significant (P = 0.013, test)3.Experiment of cell-mediated immunity, in accordance with hydrochloric acid concentration nolatrexed incremental administration, a significant increase in apoptotic index, that is, the rapid reduction of tumor cells, and Bcl-2 positive rate decreased significantly. When the drug reaches the maximum when the concentration (200μg/ml), apoptotic index up to 48.95±3.32, while the Bcl-2 positive expression rate of a minimum of 10.62±1.21, compared with the control group P <0.01, with statistical significance. Hydrochloric acid showed that in vitro nolatrexed of HepG2 human hepatoma cell proliferation inhibition are strong, can promote tumor cell apoptosis, reduced apoptosis inhibitory gene Bcl-2 expression.Conclusion: 1.Nolatrexed hydrochloride on human hepatocellular carcinoma HepG-2 with the significant growth inhibition, and its single-drug group was significantly higher than the rate of growth inhibition ADM group and DDP group, and the inhibitory rate of high-dose group was significantly higher than other dose groups, has a good dose-effect relationship.2.Nolatrexed hydrochloride on the in vitro proliferation of HepG-2 hepatoma cells have a stronger inhibitory effect can be reduced inhibitor of apoptosis Bcl-2 gene expression and promote apoptosis of tumor cells.