摘要
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。
To understand whether K562 cell was able to produce the acquired resistance to verapamil(VER),a model of K562 cell subline resistance to VER(K562/VER) was established through gradually increasing VER concentration in the media.MTT,HPLC and S-P immunocytochemistry (ICC) technique were used to detect the drug resistance to VER,dipyriamole(DPM) and cyclosporin A(CsA) in K562 and K562/VER cells,intracellular accumulation of VER,the expression of P-glycoprotein,(PgP),GST,oncoproteins and tumor suppressor proteins respectively.As compared with a K562 cells,there was a 10.4-times resistance to VER in K562/VER cells,a decreased 3.5 times～4.5 times in intracellular accumulation of VER in K562/VER cells.The expressions of middle level or high level of P 53 ,P 16 ,c-fos and low level of P 21 were observed in both K562 cells and K562/VER cells,but not c-myc and c-erbB-2.K562/VER cells expressed high level of PgP,Bcl-2 expression was found only in K562 cells.These findings suggest that K562 cell line has a capability of inducing acquired resistance to VER,which may be associated with overexpression of PgP,and then leading to reduction of intracellular accumulation of VER in K562/VER cell subline.
出处
《中国医师杂志》
CAS
1999年第6期14-16,共3页
Journal of Chinese Physician
关键词
白血病
维拉帕米
耐药性
基因表达
Human leukemic cell Verapamil Drug resistance Expression of genes
