论文部分内容阅读
为探讨人白血病细胞对耐药逆转维拉帕米的耐药机制，我们通过不断提高培养液中维拉帕米浓度建立维拉帕米耐药性Ｋ５６２亚系（Ｋ５６２／ＶＥＲ），采用ＭＴＴ法测定维拉帕米、潘生丁和环孢素Ａ对Ｋ５６２、Ｋ５６２／ＶＥＲ细胞的毒性或耐药性；采用高效液相色谱仪、荧光检测器检测细胞内维拉帕米聚集量；采用Ｓ—Ｐ免疫细胞化学技术检测Ｐ—糖蛋白（ＰｇＰ）、ＧＳＴ、癌基因蛋白和抑癌基因蛋白的表达。结果显示：Ｋ５６２／ＶＥＲ细胞对ＶＥＲ的耐药程度是Ｋ５６２细胞的１０４倍，并对潘生丁和环孢素Ａ存在交叉耐药性；Ｋ５６２／ＶＥＲ细胞内ＶＥＲ聚集量比Ｋ５６２细胞下降３５～４３倍，二者均中度或高度表达Ｐ５３、Ｐ１６、ｃ—ｆｏｓ蛋白，低度表达Ｐ２１蛋白；Ｋ５６２／ＶＥＲ细胞高度表达ＰｇＰ，Ｂｃｌ－２只在Ｋ５６２细胞中表达。说明Ｋ５６２细胞系能产生对ＶＥＲ的获得性耐药性，其耐药机制可能与ＰｇＰ表达过量，并由此导致细胞内ＶＥＲ聚集下降等因素有关。
In order to investigate the resistance mechanism of human leukemia cells to drug resistance reversed verapamil, we established the verapamil-resistant K562 subline (K562/VER) by continuously increasing the concentration of verapamil in the culture fluid, using the MTT method. To determine the toxicity or resistance of verapamil, dipyridamole and cyclosporine A to K562 and K562/VER cells; to detect the concentration of verapamil in the cells by high performance liquid chromatography and fluorescence detector; to use S-P The expression of P-glycoprotein (PgP), GST, oncogene protein and tumor suppressor gene protein was detected by immunocytochemistry. The results showed that K562/VER cells were 10 to 4 times more resistant to VER than K562 cells, and there was cross-resistance to dipyridamole and cyclosporine A. The concentration of VER in K562/VER cells was lower than that of K562 cells by 3%. The expression of P53, P16, and c-fos proteins was moderately or highly expressed, and P21 protein was lowly expressed. The K562/VER cells highly expressed PgP, and Bcl-2 was only expressed in K562 cells. It shows that K562 cell line can produce acquired resistance to VER, and its resistance mechanism may be related to the excessive expression of PgP, which leads to the decrease of intracellular VER aggregation and other factors.