Yu G.,Soochow University of China |
Yu G.,Tumor Immunology and Gene Therapy Research Center |
Jing Y.,Tumor Immunology and Gene Therapy Research Center |
Kou X.,Soochow University of China |
And 3 more authors.
Chinese Journal of Cancer Biotherapy | Year: 2013
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 (22. 34 ± 1. 12)% and (26. 78 ±1. 56)%; those in the HSC-CM + cisplatin group were (17. 22 ± 1. 42)% and (21. 52 ± 1. 76)% 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 ± 2. 89) % 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.