GRP94表达在肺癌细胞对VP-16化疗耐药中的作用

Expression of GRP94 and its function in chemotherapy resistance to VP-16 in human lung cancer cell line SPCA1

关键词:  GRP94  人类肺腺癌系  VP-16耐药

Key words:  GRP94  human lung adenocarcinoma  VP-16 resistance

摘要:

［目的］检测在Ca2+拮抗剂A23187诱导下，人肺腺癌细胞SPCA1中GRP94的表达水平，并分析其表达在肺癌细胞对VP-16耐药中的作用。［方法］采用RT-PCR、免疫荧光细胞化学方法检测不同浓度A23187处理组细胞的GRP94核酸及蛋白表达，用MTT法测定细胞在VP-16作用下的生存率。［结果］A23187可明显诱导SPCA1细胞GRP94核酸和蛋白表达，且表达水平随A23187呈浓度依赖性增加。MTT法测定结果显示：诱导组细胞（A23187浓度为0.5～6 μmol/L）对VP-16的IC50值明显高于对照组，而且IC50值的升高亦对A23187呈一定的浓度依赖性，当A23187浓度为6 μmol/L时 IC50值最大。［结论］GRP94的表达与肺腺癌细胞SPCA1对VP-16的耐药性有关，有针对性地抑制GRP94基因表达或功能可能成为肺癌治疗的新方法。

Abstract:

［Objective］To investigate the expression of GRP94 under the induction of A23187 and its function in drug resistance to VP-16 in human lung cancer cell line SPCA1.［Methods］RT-PCR and Immunofluorescence were employed in analyzing the expression of GRP94 at mRNA and protein in SPCA1 cells treated with A23187 at different concentration (0,0.5,1.0,1.5,2.0, 4.0,6.0 μmol/L).Cell survival rate following VP-16 treatment was determined with MTT assay.［Results］To a certain extent,the expression of GRP94 at both mRNA and protein level was significantly,exhibiting up-regulated by A23187 a dose-dependent manner.The mRNA expression of GPR94 was elevated up to 0.7~4.3 folds,compared to the control.The IC50s to VP-16 also showed a substantial elevation for the cells induced by A23187 at different concentrations.Survival curves analysis showed that the A23187 induction caused a significant prolonged survival for the cells subjected to VP-16 (P<0.05).［Conclusions］Up-regulated GRP94 is associated with the chemoresistance of SPCA1 cells to VP-16.