[运用SYBR实时荧光定量PCR技术检测肝癌细胞中多药耐药相关蛋白MRP2、MRP3及MRP5的mRNA表达]
[Detection of multidrug resistance-associated proteins MRP2, MRP3, and MRP5 mRNA expressions in hepatocarcinoma cells using SYBR real-time PCR].
作者信息
Zhang Lei, Fang Chi-Hua, Fan Ying-Fang
机构信息
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2008 Feb;28(2):219-21.
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.
目的
研究编码多药耐药相关蛋白2（MRP2）、MRP3和MRP5的基因在人正常肝细胞系L-02、肝癌细胞系BEL及其阿霉素耐药细胞系BEL/ADM中的差异表达。
方法
分别从L-02、BEL和BEL/ADM细胞中提取总RNA，并将mRNA逆转录为cDNA。采用实时荧光定量PCR根据标准曲线检测MRP2、MRP3、MRP5 mRNA的表达水平。
结果
MRP2 mRNA在BEL/ADM细胞系中的表达显著高于L-02和BEL细胞（P=0.000），而后两者细胞中MRP2 mRNA的表达水平相似（P=0.468）。MRP3和MRP5 mRNA的表达在这3种细胞系之间存在统计学显著差异（P=0.000）。
结论
MRP2可能在固有耐药中起作用，而MRP3和MRP5与肝癌的获得性耐药有关。
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