The Influence Of Laminaria Polysaccharidesulafate (LPS) On Lovo Cell's Generation And Apoptosis

Objective : Colorectal cancer is one of the common gastrointestinal neoplasms, and its incidence rate is rising year by year. Although the different available therapies, which including surgical operation ,radical therapy, and chemical therapy , have currently been used in the treatment of colorectal cancer, and all of these have markedly improved the prognosis of colorectal cancer. But results are not satisfied. As for the patients visiting, about 15%～35% already have tumor metastasis because there are not overt symptoms in the early period of colorectal cancer, and tumor metastasis is one of the majority reasons to result in death.At the end of 70s, it is found that the metastasis of tumor was relativity with a sore of substance like heparin. Although these phenomena well-documented, it has takes 20years to isolate a heparanase gene, mainly because of instability in designing specific, quantitative assays. Cloning and functional characterization of the long sought-after heparanase opens a new chapter in the understanding and potential manipulation of metastasis processes.Laminaria polysaccharidesulfate(LPS) is a watersolibility polysaccharide which include many types of monosaccharide and sulphuricacid. It's biological function mainly concentrate on antitumor ,adjusting immunity, regularing blood fat and blood sugar and so on. Our country produced most laminaria japonica of the world. Laminaria japonica can eat and also can be used as medicine. It is reported that LPS can inhibit the transfusion of tumor cells in animal exprements. There is also some studying reported that LPS can inhibit the generation of tumor cells in vivo or in vitro, but it's molecular organism biomechanism have not reported. Our item study that :LPS enhance the apoptosis of tumor cells ,inhibit the generation of the cells and enhance the sensitivity of chemical medicine. Bcl-2 which has an intimate relation with the progressing of tumor is an anti- apoptosis gene . Bcl-2 gene play it's function through it's transcript which called bcl-2 gene protein. Bcl-2 gene protein participate endo-cell anti-oxygen and inhibit the progress of Ca across memberance movement to adjust the apoptosis of tumor cells. We analysis cell cycle's change and detect bcl-2 gene and NF-κBp65 gene in order to observe the antitumor mechanism of LPS.Methods: 1 We use different density of LPS to deal with lovo cells of carcinoma of human large intestine. We use MTT method to detect cells'generation and use cell staining method to detect the apoptosis shape of cells.2 We use flow cytometry method (FCM) to detect the change of cell cycles and the expression of bcl-2 gene and NF-κBp65 gene.3 We use RT-PCR to detect the expression of heparanase. The purpose of all study methods are to study the influence on cell's generation, apoptosis and cell cycles of lovo cells which belong to a type of carcinoma of human large intestine. At last use statistic method to analyze all data.Results: 1 Morphology observation: we observe the cells of comparison groups and study groups through Scanning electron microscopy(SEM): the generation of cells in comparison groups is active,cells are in different shapes, cells have little cytoplasm ,cell's membrane is ambiguity and nuclear membrane is integrated. Cell's chromatin is thick and we also can observe caryocinesia phase.The study groups: it is great different in cells'shapes. Degeneration and necrosis have take placed in many cells. Decomposition have take placed in cytolymph. Cells'nuclear have broken up ,deliquesced or even disappeared.2 We use MTT method to detect the inhibit condition of LPS on lovo cells. With the time extending and the increasing of the LPS'density , the inhibit function become stronger (P<0.05).3 Cell cycle's change: We use PI to deal with the study groups'cells and we find that LPS can change lovo cell'cell cycle in large extent.4 The expression of bcl-2 gene and NF-κBp65 gene: LPS can lower the expression of bcl-2 gene and NF-κBp65 gene. The more increasing of LPS'density the lower the expression of those two genes.Conclusions: 1 LPS can inhibit the generation of lovo cells and can enhance the sensitivity of chemical medicine.2 LPS can down regular the expression of bcl-2 gene and NF-κBp65 gene. Through this method, LPS also can lower the inhibition to lovo cells.3 LPS can obviously inhibit the function of heparanase and tumor transfusion indirectly.