目的 探讨三磷酸腺苷结合转运蛋白G家族成员2（ATP-binding cassette superfamily G member 2,ABCG2）抑制剂维拉帕米对体内外胰腺癌SW1990细胞侵袭转移能力影响及其机制。方法 以终浓度分别为0、12.5、25、50、100和200μmol/L维拉帕...目的 探讨三磷酸腺苷结合转运蛋白G家族成员2（ATP-binding cassette superfamily G member 2,ABCG2）抑制剂维拉帕米对体内外胰腺癌SW1990细胞侵袭转移能力影响及其机制。方法 以终浓度分别为0、12.5、25、50、100和200μmol/L维拉帕米处理SW1990细胞24、48和72h后,以CCK-8法检测维拉帕米对SW1990细胞增殖抑制影响;RT-PCR和蛋白质印迹法检测维拉帕米对体内外SW1990细胞ABCG2mRNA及蛋白表达水平的影响;Transwell小室侵袭迁移实验及划痕实验分析维拉帕米处理后细胞侵袭迁移能力的改变。将SW1990细胞接种至裸鼠皮下,对比观察维拉帕米干扰前后肿瘤细胞在裸鼠体内的成瘤情况;免疫组化分析裸鼠肿瘤组织中ABCG2的表达。结果 CCK-8检测结果显示,浓度为25~100μmol/L维拉帕米对胰腺癌SW1990细胞的抑制呈现明显的剂量及时间依赖性。划痕实验结果显示,细胞平均迁移率比较,划痕24h维拉帕米组为（19.2±2.04）%,对照组为（36.8±2.25）%,t=-17.23,P〈0.001;48h维拉帕米组为（43.7±3.14）%,对照组为（78.4±2.67）%,t=-23.85,P〈0.001。Transwell侵袭实验结果显示,维拉帕米组平均穿膜细胞数为46.6±3.3,明显少于对照组的90.2±2.7,t=-47.2,P〈0.001;迁移实验结果显示,维拉帕米组平均穿膜细胞数为61.4±2.8,亦少于对照组的110.3±3.5,t=-39.5,P〈0.001;蛋白质印迹法及RT-PCR结果显示,维拉帕米能够明显降低体内外SW1990细胞ABCG2蛋白及mRNA的表达水平。裸鼠成瘤实验结果显示,细胞接种后10d左右可见肿瘤结节,50d时维拉帕米组裸鼠肿瘤体积为（521.6±48.5）mm3,小于对照组的（1 496.6±73.1）mm3,t=-38.6,P〈0.001;维拉帕米组瘤质量（0.53±0.18）g,明显低于对照组（1.61±0.45）g,t=-22.49,P〈0.001。免疫组化结果显示,维拉帕米能明显降低SW1990细胞ABCG2的表达。结论 维拉帕米能明显抑制胰腺癌SW1990细胞在体内外的侵袭和转移,其机制可能与维拉帕米下调ABCG2的�展开更多
Objective To identify effects of bile acids on pancreatic cancer, The ultrastructure and growth of PANC-1 and MIA PaCa-2 cell lines in crude bile modified medium were studied. Methods The growth of PANC-1 and MIA PaCa...Objective To identify effects of bile acids on pancreatic cancer, The ultrastructure and growth of PANC-1 and MIA PaCa-2 cell lines in crude bile modified medium were studied. Methods The growth of PANC-1 and MIA PaCa-2 cells in RPMI 164o with or without 1%, 2% and 4% of the purified crude bile (containing total bile acids 1o. 17mmol/L) was assessed for 2, 4, 6, 8d by using MTT assay to determine inhibitory rate- The cell surface and intracellular ultrastructure of PANC-1 cells was investigated by SEM and TEM at 24h and 48h, respectively. Results The proliferation of both cell lines in bile treated medium were greatly retarded (P <o. oo1). The inhibitory rate of 1 %, 2% and 4% bile on Panc-1 cells in 4d were 38%, 6o% and 66%, respectively (P <o. o5), on MIA PaCa-2 cells at 4d were 28%, 39% and 52%, respectively (P <o. o5). The ceIls grown in bile for 48h lost their microvilli, their mitochondria and other organelles became vacuolated. Conclusion The bile acids in bile has cytotoxicity on PANC-1 and MIAPACA-2 cells, which may inhiblt pancreatic cancer progress in patients clinically.展开更多
文摘Objective To identify effects of bile acids on pancreatic cancer, The ultrastructure and growth of PANC-1 and MIA PaCa-2 cell lines in crude bile modified medium were studied. Methods The growth of PANC-1 and MIA PaCa-2 cells in RPMI 164o with or without 1%, 2% and 4% of the purified crude bile (containing total bile acids 1o. 17mmol/L) was assessed for 2, 4, 6, 8d by using MTT assay to determine inhibitory rate- The cell surface and intracellular ultrastructure of PANC-1 cells was investigated by SEM and TEM at 24h and 48h, respectively. Results The proliferation of both cell lines in bile treated medium were greatly retarded (P <o. oo1). The inhibitory rate of 1 %, 2% and 4% bile on Panc-1 cells in 4d were 38%, 6o% and 66%, respectively (P <o. o5), on MIA PaCa-2 cells at 4d were 28%, 39% and 52%, respectively (P <o. o5). The ceIls grown in bile for 48h lost their microvilli, their mitochondria and other organelles became vacuolated. Conclusion The bile acids in bile has cytotoxicity on PANC-1 and MIAPACA-2 cells, which may inhiblt pancreatic cancer progress in patients clinically.
