目的：探讨肝星状细胞条件培养基（hepatic stellate cell conditioned medium，HSC-CM）对人肝癌PLC/PRF/5细胞耐药性的影响及其可能的机制。 方法： 用无血清RPMI 1640培养肝星状细胞LX-2，使其在缺营养的环境下活化，收集其条件培养上清即为HSC-CM。PLC/PRF/5细胞在HSC-CM条件下培养24 h后，顺铂处理12 h或24 h，采用流式细胞术检测PLC/PRF/5细胞的凋亡情况，MTT法检测PLC/PRF/5细胞的增殖，real-time PCR检测PLC/PRF/5细胞上皮间质转化（epithelial mesenchymal transition，EMT）相关基因的表达水平。 结果： 顺铂组12和24 h两个时间点PLC/PRF/5细胞的凋亡率为（22.34±1.12）%和（26.78±1.56）%；HSC-CM+顺铂组细胞的凋亡率为（17.22±1.42）%和（21.52±1.76）%，顺铂组细胞凋亡率显著高于HSC-CM+顺铂组（P<0.05）。同在这两个时间点，顺铂组和HSC-CM+顺铂组PLC/PRF/5细胞的增殖活性分别为（68.65±2.56）%和（79.47±1.43）%，（46.54±3.65）%和（62.77±2.89）%，HSC-CM+顺铂组细胞增殖活性均高于顺铂组（P<0.05）。Real-time PCR 结果显示，与顺铂组相比较，HSC-CM+顺铂组PLC/PRF/5细胞中上皮标记物钙黏蛋白（E-cadherin）的表达下降（P<0.05），而间质细胞标记物神经黏附素（N-cadherin）、波形蛋白（vimentin）以及EMT相关转录因 子Snail和ZEB1的表达显著上调（P<0.01）。 结论: HSC-CM可能通过诱导PLC/PRF/5细胞发生EMT，从而增强PLC/PRF/5 细胞对顺铂的抵抗作用。

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.

国家自然科学基金资助项目（No. 31171321，No. 81201584）；上海市自然科学基金资助项目（No. 2ZR1439800，No. 12ZR1454200）