实时荧光定量PCR检测肝癌细胞多药耐药相关蛋白MRP2、MRP3及MRP5 mRNA的表达

目的 研究多药耐药相关蛋白2(MRP2)基因及MRP3、MRP5基因在正常肝脏细胞系L-02、肝癌细胞系BEL及肝癌耐药细胞系BEL/ADM的表达差异,并探讨其在肝癌耐药中的意义.方法 分别提取L-02、BEL及BEL/ADM细胞的总RNA,利用实时荧光定量PCR法检测MRP2、MRP3、MRP5基因的表达水平.结果 MRP2基因在BEL/ADM细胞中的表达量明显高于在L-02、BEL细胞中的表达(P=0.000),而其在L-02、BEL两种细胞中的表达没有显著性差异(P=0.468);MRP3、MRP5基因表达量在L-02、BEL、BEL/ADM三种细胞中均有显著性差异(P=0.000).结论 MRP2可能参与了肝癌的内在性耐药,而MRP3、MRP5可能与肝癌的获得性耐药有关.

Detection of multidrug resistance-associated proteins MRP2,MRP3,and MRP5 mRNA expressions in hepatocarcinoma cells using SYBR real-time PCR

OBJECTIVE: To investigate the differential expression of genes encoding multidrug resistance-associated protein 2 (MRP2), MRP3 and MRP5 in human normal liver cell line L-02, hepatocarcinoma cell line BEL and its adriamycin-resistant counterpart BEL/ADM cell line. METHODS: The total RNA was extracted from L-02, BEL and BEL/ADM cells, respectively, and the mRNAs were reversely transcribed into cDNA. The expression levels of MRP2, MRP3, MRP5 mRNAs were detected according to the standard curve using real-time fluorescence quantitative PCR. RESULTS: The expression of MRP2 mRNA in BEL/ADM cell line was significantly higher than that in L-02 and BEL cells (P=0.000), and the latter two cells showed similar expression levels of MRP2 mRNA (P=0.468). The expressions of MRP3 and MRP5 mRNA showed statistically significant difference between the 3 cell lines (P=0.000). CONCLUSION: MRP2 might play a role in the intrinsic drug resistance, and MRP3 and MRP5 are related to the acquired drug resistance of hepatocellular carcinoma.