Yan Feng, Wang Xiao-Min, Pan Chao
Postdoctoral Station, Zhongshan Hospital, Xiamen University, Xiamen, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Mar;29(3):483-6.
To detect the expression of extracellular-signal regulated protein kinase 5 (ERK5) in multidrug-resistant hepatocellular carcinoma cell lines hepG2/ADM and BEL-7402/5-FU and in the parental cell lines (hepG2 and BEL-7402), and study the role of ERK5 in multidrug resistance of hepatocellular carcinoma.
MTT assay was used to determine the multidrug resistance (MDR) of hepG2/ADM and BEL-7402/5-FU cells. Real-time PCR and Western blotting were used to detect the expression of ERK5 at mRNA and protein levels in the 4 cell lines.
The resistance indices to ADM, 5-FU, and CDDP was 12.34, 5.74, and 3.81 in hepG2/ADM cells, and the resistance indices to 5-FU, VCR, OHP, MTX, and ADM was 15.32, 10.08, 5. 85, 6.74 and 3.26 in BEL-7402 cells, respectively. Compared with those in hepG2 and BEL-7402 cells, the ERK5 gene was up-regulated at both mRNA and protein levels in HepG2/ADM cells, but down-regulated at the mRNA level and up-regulated at the protein level in BEL-7402/5-FU cells.
ERK5 is related to multidrug resistance in HepG2/ADM and BEL-7402/5-FU cells, and may provide new clues for reversing multidrug resistance of hepatocellular carcinoma cells.
检测细胞外信号调节蛋白激酶5(ERK5)在多药耐药肝癌细胞系hepG2/ADM和BEL-7402/5-FU及其亲本细胞系(hepG2和BEL-7402)中的表达,并研究ERK5在肝癌多药耐药中的作用。
采用MTT法检测hepG2/ADM和BEL-7402/5-FU细胞的多药耐药性。采用实时荧光定量PCR和蛋白质印迹法检测4种细胞系中ERK5在mRNA和蛋白质水平的表达。
hepG2/ADM细胞对阿霉素(ADM)、5-氟尿嘧啶(5-FU)和顺铂(CDDP)的耐药指数分别为12.34、5.74和3.81;BEL-7402/5-FU细胞对5-FU、长春新碱(VCR)、奥沙利铂(OHP)、甲氨蝶呤(MTX)和ADM的耐药指数分别为15.32、10.08、5.85、6.74和3.26。与hepG2和BEL-7402细胞相比,HepG2/ADM细胞中ERK5基因在mRNA和蛋白质水平均上调,而BEL-7402/5-FU细胞中ERK5基因在mRNA水平下调,在蛋白质水平上调。
ERK5与HepG2/ADM和BEL-7402/5-FU细胞的多药耐药有关,可能为逆转肝癌细胞的多药耐药提供新的线索。