目的 通过干扰RNA的方法下调人磷酸葡萄糖异构酶/自分泌因子（PGI/AMF）基因的表达，研究人PGI/AMF基因协同人参皂苷Rh₂对白血病KG1α细胞的影响。方法 将对数生长期KG1α细胞及转染细胞分成对照组和用药组，对照组常规培养，药物组细胞培养体系中分别加入30、45、60、75、90 μmol/L的人参皂苷Rh₂，各组分别培养24、48、72 h后，CCK-8检测人参皂苷Rh₂对白血病KG1α及转染株细胞增殖的影响；台盼蓝检测人PGI基因对KG1α细胞增殖的影响；Antibody Array检测人参皂苷Rh₂对Akt通路蛋白的影响；Western blotting检测下调PGI/AMF与人参皂苷Rh₂在mTOR、Raptor、Rag蛋白表达变化方面的相关性。结果 人参皂苷Rh₂抑制KG1α的增殖；下调PGI/AMF使KG1α对人参皂苷Rh₂更加敏感；下调PGI基因抑制细胞增殖；Antibody Array结果显示人参皂苷Rh₂通过下调P38、mTOR、Akt、AMPKα、PARP、Bad的表达影响KG1α增殖；Western blotting结果显示下调PGI基因通过抑制mTOR、Rag、Raptor表达与人参皂苷Rh₂产生协同作用调节人白血病细胞增殖。结论 下调PGI基因协同人参皂苷Rh₂抑制KG1α细胞的增殖。

Objective To investigate the down-regulation of phosphoglucose isomerase / autocrine motility factor (PGI/AMF) gene expression by interfering RNA for study on the pharmacological effect of PGI/AMF with ginsenoside Rh₂ on leukemia KG1α cells. Methods The KG1α and the silencing PGI/AMFα KG1α (siPGI-KG1α) cells at logarithmic growth phase were divided into control and drug groups in different dosages. The cells in the control group were normally treated and the cells in the drug groups were incubated with ginsenoside Rh₂ in the concentration of 30, 45, 60, 75, and 90 μmol/L in 24, 48, and 72 h respectively. Cell Counting Kit-8 (CCK-8) was used to demonstrate the effect of ginsenside Rh₂ on proliferation of KG1α and siPGI-KG1α cells. Trypan blue staining was applied to detecting the effect of human PGI on the proliferation of leukemia KG1α cell. The changes of KG1α by ginsenoside Rh₂ on Akt pathway were tested by Antibody Array. The corelation of down-regulating PGI/AMF and Rh₂ about mTOR, Raptor, and Rag was detected by Western blotting. Results Compared with the control group, ginsenoside Rh₂ had significant inhibition on the proliferation of KG1α. Down-regulation of PGI/AMF could make KG1α more sensitive to ginsenoside Rh₂ and down-regulation of PGI could inhibit the proliferation of KG1α. Antibody Array showed that ginsenoside Rh₂ could inhibit the proliferation of KG1α cells through decreasing the expression of P38, mTOR, Akt, AMPKα, PARP, and Bad. Western blotting results indicated that down-regulation of PGI through the synergy of mTOR, Rag, and Raptor with ginsenoside Rh₂ could inhibit the proliferation of KG1α cells. Conclusion Down-regulation of PGI/AMF coordinated with ginsenoside Rh₂ could inhibit the proliferation of KG1α cells.

国家自然科学基金面上项目（31271368）；重庆市教委基金项目（KJ110328）