Study On The Effect And Mechanism Of Hirsutella Sinensis Polysaccharide On Lung Cancer

Polysaccharide is the most important bioactive component in Hirsutella sinensis. It has a very good application prospers because of its little toxic effect on tissue cells. This paper focused on the HSP-III, which is a kind of fungus polysaccharide with good biological property. HSP-III like the effect of HSP-III on lung adenocarcinoma cell (H1299)proliferation, apoptosis and the migration ability in vitro.The strain was activated through the PDA solid culture medium, and the culture of the Hirsutella sinensis was expanded by liquid fermentation. By using hot water extraction and ethanol precipitation method to obtain the crude hirsutella polysaccharide (HSP- 1); HSP-Ⅱ was obtained by removal of protein using Sevage method, HSP- Ⅲwas obtained using Sephadex G-100 column chromatography, yields were 12.60%, 5.43% and 0.93%. MTT method was used to detect the proliferation of H1299 cells in vitro. The results showed that HSP-Ⅲ has an inhibitory effect on the H1299 cells proliferation. At the molecular level by immunofluorescence, Western blot and RT-PCR test cell proliferation marker PCNA protein, the experimental results show that the proliferation of H1299 cells was inhibited with the HSP-Ⅲ effect. The apoptosis of H1299 cells was detected by TUNEL method and the mitochondrial membrane potential of JC-1 cells. The results showed that the H1299 cells apoptosis was significantly increased. It showed that the apoptosis degree was significantly increased in the case of H1299 cells with HSP-Ⅲ. In the experiment for of the cell migration ability of HSP-Ⅲ H1299 was detected by HSP-Ⅲ. The experimental results showed that there is no significantly different shortening of the cell line, and the numbers of cells are basically the same through the membrane vesicles. It can be explained that, the migration ability of H1299 cells is not affected.In the present study, Rh123 and blot Western were used to verify the mitochondria of H1299 cells. The results showed that when the effect of HSP-Ⅲ on H1299 cells, the mitochondrial membrane potential was significantly decreased, Bax protein was increased,protein content of cytochrome C in the cytosol was increased and caspase protein family was activated. The mechanism was shown in Figure 4.In order to verify the HSP-III on H1299 cells through the mitochondrial apoptosis pathway, the experiment also uses DCFH-DA molecular probe and Blue PAGE Native in ROS to detect the change of respiratory chain complexes in mitochondria. The experimental results show that when HSP-Ⅲ is acting on H1299 cells.In this experiment, intratumoral injection was taken to simulate the tumor growth, and then the effect of HSP-Ⅲ on the tumor was evaluated by HSP-Ⅲ subcutaneously of intradermal. The results showed that the growth rate of the tumor was slow, the tumor size was decreased and the weight of the tumor decreased under the HSP-Ⅲ treatment. It is proved that HSP-Ⅲ inhibits the growth of H1299 cells in vivo.