The Effect Of Ruanjiansanjie On Inhibition Of SGC-7901 Proliferation By VEGF-ERKsignal-transduction Pathway

Objective:The experiment using serum drug pharmacology method adopt CCK8, confocal microscopy, flow cytometry, Western Blot and RT-PCR technical means to research Ruanjiansanjie on the cell proliferation and apoptosis of gastric cancer SGC7901.Methods:1.Preparing the medicated serum of Ruanjiansanjie.2.Testing the effects of Ruanjiansanjie serum containing serum on SGC7901 cells.Detecting the proliferation of serum containing on gastric cancer SGC7901 cells by CCK8 method.3.Apoptosis is determined by Confocal microscopy, FCM with Annexin V FITC and propidium(PI) staining.4.Exploring the effects and mechanisms of Ruanjiansanjie serum containing:using the method of Western Blot to detect the influence on Ruanjiansanjie serum on ERK,phosphorylation of p-ERK and VEGF expression;use the method of RT-PCR to detect the mRNA expressions of the serum of Ruanjiansanjie on Caspase-3,Bax,and Bcl-2 in gastric cancer cells.Results:1 Ruanjiansanjie could obviously inhibit proliferation of gastric cancer cell and appear dose-dependent effects(P<0.05).2 Under the Confocal Laser Scanning microscopy showed that Medicated serum cell apoptosis occurred after serum containing drug treatment: the negative control group with normal activity of cell-based, Annexinv were low and PI staining, membrane integrity,cellular morphology normal; low-dose group were observed in high Annexin Vstaining,and PI low staining, suggesting that some cells were in the early stages of apoptosis, cell morphology was still normal; middle dose group were observed in serum containing high Annexinv dye and PI low staining, suggesting that part of the late apoptotic cells and cell morphology round; high-dose group containing serum PI high staining, appear fraction of cells membrane rupture, nuclei were irregular fragments ranging in size shape, typical apoptotic morphology, suggesting that high-dose group were seen a large number of advanced cell.3 Flow cytometry showed that when gastric cancer SGC-7901 cells were treated with Ruanjiansanjie, the SGC-7901 cell apoptosis rate increased(P<0.05).4 Western Blot analysis showed that when gastric cancer SGC-7901 cells were treated with Ruanjiansanjie, p-ERK expression was significantly decreased(P<0.05),VEGF expression was significantly increased(P<0.05).5 RT-PCR analysis showed that when gastric cancer SGC-7901 cells were treated with Ruanjiansanjie capsule, Caspase-3 and Bax m RNA expression was significantly increased(P<0.05),Bcl-2 expression was significantly decreased(P<0.05).Conclusion:1.Ruanjiansanjie can significantly inhibit the proliferation of human gastric cancer SGC-7901 cells and induce apoptosis in human gastric cancer.2.Ruanjiansanjie can significantly inhibit the growth of human gastric cancer SGC-7901 cells.The main mechanism is the inhibition of VEGF and p-ERK expression,thereby inhibiting their proliferation and promoting apoptosis;at the same time Ruanjiansanjie can induce apoptosis by increasing the expression of Bax and Caspase-3and decreaseing the expression of Bcl-2.