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Effect of ginsenoside Rh_2 on proliferation and apoptosis of KG1α cells by autophagy pathway

LIU Xiao-xia;CHEN Yi;XIONG Wei;LI Jing;CHEN Di-long;LIU Ze-hong;Traditional Chinese Medicine Department,Jiangjin Central Hospital of Chongqing;Laboratory of Stem Cell and Tissue Engineering,Chongqing Medical University;Department of Physiology,Chongqing Medical and Pharmaceutical College;  
Objective To investigate the antitumour activity of ginsenoside Rh_2 against human leukemia KG1α cells through apoptosis and autophagy pathway. Methods CCK-8 assay was used to screen the most effective ingredient on the proliferations among ginsenoside Rh_2 in leukemia KG1α cell line; FCM detected cell apoptosis; Hoechst staining observed the cell morphological changes of apoptosis; Acridine staining detected Rh_2 effected on autophagy; Western blotting and RT-PCR detected the expression levels of the proteins closely associated with autophagy and apoptosis. After joining autophagy inhibitors, using CCK-8 to test the proliferation activity of cells, cell apoptosis was measured by FCM. Results CCK-8 indicated that Rh_2 could inhibit the proliferation of KG1α cells significantly with dose- and time-dependent manners; FCM indicated that Rh_2 induced apoptosis; Hoechest staining showed that KG1α cells had typical apoptotic morphological changes by treated Rh_2; Acridine staining revealed that Rh_2 cause increase in the number of acidic autophagy vesicles in cells, causing cell autophagy levels increased; Western blotting and RT-PCR results showed that Rh_2 increased the expression of Beclin-1, LC3 A, and LC3 B, activated Caspase-3 and Bax/Bcl-2 rates, and MAPK, ATK, and ERK signaling pathway; After using autophagy inhibitors(3-MA), autophagy crippled that Rh_2 inhibited the proliferation and induced apoptosis in KG1α cells. Conclusion Ginsenoside Rh_2 could significantly enhance autophagy through activated MAPK, ATK, and ERK signaling pathway, and then inhibit the proliferation and induce apoptosis in KG1α cells.
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