Effect Of Sijunzitang On Gastric Cancer Side Population Cell Proliferation, Cycle And Apoptosis

Objective:Starting from the angle of side population cells in gastric cancer study decoction of four noble drugs on gastric cancer cell proliferation,and apoptosis, As the step of Sijunzitang action research group to research mechanism of gastric cancer and gastric cancer side population cells lay the foundation.Method:1.Human gastric cancer cell line highly differentiated gastric cancer cell lines MGC-803, moderately differentiated gastric cancer cell line SGC-7901and poorly differentiated gastric cancer cell line BGC-823in vitro Cultured;2. Preparation of serum of Sijunzitang;3. Cytometry and sorting of three gastric cancer cell lines by flow side population cells;4.Application of CCK-8assay understand the drugs Sijunzitang serum interventions before and after the intervention of SP cells and NSP cells and unsorted cell proliferation ability;5. Flow cytometry is used to test drug serum SP cell cycle and apoptosis of change after the intervention;6. Using Western blot method to detect drug serum after the intervention of SP cells apoptosis proteins BAX and Bcl-2expression changes.Results:2. SP cells proliferation multiples of7days (SGC-7901:20.90±0.66; BGC823:29.50+±0.64) significantly higher than the NSP cells and no separation cells(P＜0.01).3. With the increase of drug serum concentration and action time prolonged, the proliferation of SP cells was inhibited more obvious. SGC-7901SP cells proliferation inhibition degree to low doses:45.87±0.10%, medium doses:64.36±1.18%, high doses:73.58±2.48%; BGC-823SP cells proliferation inhibition degree to low doses:41.45±0.11%, middle dose:57.80±3.24%, high doses:69.85±0.81%and NSP cells have no difference (P>0.05)4. With the increase of drug serum concentration and longer action time, SP cells in the G2phase of cell cycle significantly, cell block in G1phase. 5. With the increase of drug serum concentration and longer action time, SP cells apoptosis rate increased. SGC-7901SP cells apoptosis rate in high-dose group of48hours is28.20±2.19%; BGC-823SP cells apoptosis rate in high-dose group of48hours is19.51±2.35%were significantly higher than normal saline control group (P<0.05), and between the NSP cells and no significant difference (P>0.05).6. With elevated serum drug concentration and action time extended, SP cells promote the expression of BAX apoptosis protein increased, suppression of apoptosis protein expression of Bcl-2gradually decreased. SGC-7901SP cells BAX protein expression in high dose group of48hours to0.862±0.075; BAX protein expression BGC-823SP cells in high dose group of48hours to0.733±0.021were significantly higher than saline control group (P<0.05), and between the NSP cells and no significant difference (P>0.05). SGC-7901SP cells the Bcl-2protein expression in high dose group of48hours down to0.297±0.035; BGC-823SP cells to the Bcl-2protein expression in high dose group of48hours down to0.240±0.039were significantly lower than normal saline control group (P<0.05), and between the NSP cells and no significant difference (P>0.05).Conclusion:1. Gastric cancer cells exist SP cells; SP cells proliferation significantly faster than the speed of NSP cells and unassorted gastric cancer cells.2. Sijunzi decoction drug serum can inhibit the gastric cancer SP cells proliferation, promote gastric cancer SP cells apoptosis, promote apoptosis proteins BAX expression, reduce apoptosis proteins Bcl-2expression.3. High dose of the drug serum had no significant difference in SP and NSP cells apoptosis.