中文摘要：

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RPMI1640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24h后，顺铂处理12h或24h，采用流式细胞术检测PLC／PRF／5细胞的凋亡情况，MTF法检测PLC／PRF／5细胞的增殖，real—timePCR检测PLC／PRF／5细胞上皮间质转化（epithelialmesenchymaltransition，EMT）相关基因的表达水平。结果：顺铂组12和24h两个时间点PLC／PRF／5细胞的凋亡率为（22．344±1．12）％和（26．784±1．56）％；HSC—CM＋顺铂组细胞的凋亡率为（17．224±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（79．474±1．43）％，（46．54±3．65）％和（62．774-2．89）％，HSC-CM＋顺铂组细胞增殖活性均高于顺铂组（P〈0．05）。Real—timePCR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ZEBl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。

英文摘要：

Objective： To explore the effect of the hepatic stellate cell condition medium （HSC-CM） on chemo- resistance of human hepatoeellular 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 collect- ed, 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 ceils 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 〈 O. 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） % respeetively, showing a stronger proliferation aetivity of PLC/PRF/5 eells from the HSC-CM ＋ eispla- tin group. Real-time PCR results indicated that compared with the eisplatin group, the expression of epithelial marker E- eadherin in PLC/PRF/5 cells from the HSC-CM ＋ eisplatin group was decreased （ P 〈 0.05 ） , and the mesenehymal mark- ers （N-eadherin, vimentin） and EMT-assoeiated transeription factor （Snail, ZEB1 ） expressions were siguifieantly up- regulated （P 〈0.01 ）. Conclusion： HSC-CM may promote the rehemo-resistanee of PLC/PRF/5 ceils through inducing EMT of PLC/PRF/5 cells.