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目的探讨PDLIM4基因在正常前列腺上皮细胞(RWPE1)、前列腺癌细胞(PC-3、DU145、LNCaP)、前列腺癌多药耐药细胞(PC-3/Doc)及其他耐药细胞(K562/A02、H460/Doc)中的差异表达。方法采用定量PCR(qPCR)、Western blot、免疫荧光等方法检测PDLIM4基因在各种细胞株中的表达;利用Western blo和DNA甲基转移酶(DNMTs)活性检测分析细胞中DNMTs的表达及活性。结果 PDLIM4基因在RWPE1高表达,在DU145、LNCaP细胞中基本无表达,在前列腺癌PC-3细胞中表达甚低,而在与其对应的多药耐药PC-3/Doc细胞中高表达(P<0.05),并且在耐药细胞株中的高表达具有组织特异性。DNMTs在RWPE1表达水平低于前列腺癌细胞,但在PC-3和PC-3/Doc中无差异。酶活性分析表明,PC-3/Doc中DNMTs的活性低于PC-3细胞(P<0.05)。结论 PDLIM4基因在前列腺癌细胞中低表达,这种特异性的在前列腺癌耐药细胞中表达显著上调可能是由DNMTs活性降低所致。
Objective To investigate the expression of PDLIM4 in RWPE1, PC-3, DU145 and LNCaP, PC-3 / Doc and other drug resistant cells (K562 / A02, H460 / Doc). Methods The expression of PDLIM4 gene in various cell lines was detected by qPCR, Western blot and immunofluorescence. DNMTs expression and activity were detected by Western blotting and DNA methyltransferase (DNMT) activity assay. Results The PDLIM4 gene was highly expressed in RWPE1, but not in DU145 and LNCaP cells. The expression of PDLIM4 gene was low in PC-3 cells of prostate cancer and high in multidrug-resistant PC-3 / Doc cells (P < 0.05), and high expression in drug-resistant cell lines was tissue-specific. DNMTs expressed lower levels of RWPE1 in prostate cancer cells, but not in PC-3 and PC-3 / Doc. Enzyme activity analysis showed that DNMTs activity in PC-3 / Doc was lower than that in PC-3 cells (P <0.05). Conclusion The PDLIM4 gene is low expressed in prostate cancer cells. This specific up-regulation of prostate cancer cell lines may be due to the decreased activity of DNMTs.