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Archive > Volume 20 Issue 2 > 2013,20(2):187-191. DOI:10.3872/j.issn.1007-385X.2013.02.010 Prev Next
Basic Research
Effect of hepatic stellate cell condition medium on chemo-resistance of hepatocellular carcinoma PLC/PRF/5 cells and its mechanism
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Abstract:
Objective: To explore the effect of the hepatic stellate cell condition medium (HSC-CM) on chemo-resistance of human hepatocellular carcinoma PLC/PRF/5 cells and its possible mechanism. Methods: Hepatic stellate cell line LX-2 was incubated and activated in serum-free RPMI 1640 medium, and then this condition medium was collected, named HSC-CM. PLC/PRF/5 cells were incubated in HSC-CM for 24 h. After the treatment of cisplatin for 12 or 24 h, the apoptosis of PLC/PRF/5 cells was identified by flow cytometry, the cell proliferation of PLC/PRF/5 cells was detected by MTT assay, and the expression of epithelial mesenchymal transition (EMT) associated genes in PLC/PRF/5 cells was detected by real-time PCR. Results: The apoptosis rates of PLC/PRF/5 cells in the cisplatin group were (2234±112)% and (26.78±1.56)%; those in the HSC-CM+cisplatin group were (17.22±1.42)% and (21.52±176)% at 12 and 24 h time points, which showed that the apoptosis rates of the cisplatin group were higher than of the HSC-CM+cisplatin group (P<0.05). The proliferation of PLC/PRF/5 cells in the cisplatin group and HSC-CM+cisplatin group at these two time points were (68.65±2.56)% and （79.47±1.43)%, (46.54±3.65)% and (62.77±289) % respectively, showing a stronger proliferation activity of PLC/PRF/5 cells from the HSC-CM+cisplatin group. Real-time PCR results indicated that compared with the cisplatin group, the expression of epithelial marker E-cadherin in PLC/PRF/5 cells from the HSC-CM+cisplatin group was decreased (P<0.05), and the mesenchymal markers (N-cadherin, vimentin) and EMT-associated transcription factor (Snail, ZEB1) expressions were significantly up-regulated (P<0.01). Conclusion: HSC-CM may promote the rchemo-resistance of PLC/PRF/5 cells through inducing EMT of PLC/PRF/5 cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China （No. 31171321, No. 81201584）, and the Natural Science Foundation of Shanghai （No. 2ZR1439800, No.12ZR1454200）
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