超声辐照载多西紫杉醇微泡对人肝癌细胞MHCC97-H的促凋亡作用

The effect of ultrasound irradiating docetaxel-loaded microbubbles on apoptosis in human liver cancer cells MHCC97-H

目的探讨超声辐照载多西紫杉醇微泡对人高转移性肝癌细胞MHCC97-H凋亡的影响及其机制。方法将MHCC97-H细胞随机分为4组：对照组、载多西紫杉醇微泡组（DM）、超声辐照组（US）、超声辐照载多西紫杉醇微泡组（DM＋US），分别检测各组细胞的存活率、形态学变化、凋亡情况及丝裂原活化蛋白激酶（MAPK）通路蛋白和增殖细胞核抗原蛋白（PCNA）的表达。结果24h后，MTT法显示DM组、US组、DM＋US组细胞存活率分别为（76．0±3．0）％、（73．0±5．0）％和（52．0±6．0）％，DM＋US组降低最明显；显微镜下观察对照组肿瘤细胞形态大小正常，生长旺盛，而DM组、US组、DM＋US组细胞数量均有减少，形态不规则，细胞间隙增加，部分细胞脱落悬浮在培养液中，DM＋US组改变更明显；流式细胞仪显示DM＋US组细胞凋亡率为（26．81±2．82）％，与对照组[（2．90±0．99）％]比较差异有统计学意义（P〈0．05）；Western-blot显示DM＋US组细胞p-ERK1／2蛋白表达水平增多，p-p38蛋白表达下降，PCNA蛋白表达降低。结论超声辐照载多西紫杉醇微泡通过影响ERK1／2和p38信号转导通路，调节相关蛋白的表达而诱导人肝癌细胞MHCC97-H的凋亡。

Objective To study the effects and its probable mechanism of ultrasound-irradiation doeetaxel-loaded microbubbles on apoptosis in human liver cancer cells MHCC97-H. Methods The MHCC97-H ceils were randomly divided into 4 groups ： control group, docetaxel-loaded microbubbles group （DM）, Ultrasound group （US）, ultrasound irradiating docetaxel-loaded microbubbles group （DM ＋ US）. The livability, the morphological change of cells, cell apoptosis rate and the expression of proliferating cell nuclear antigen and MAPK pathway members of each group were respectively detected. Results After 24 h, MTT test showed that the livability of MHCC97-H cells of DM group, US group and DM ＋ US group were （76.0 ±3.0）%,（73.0± 5.0）% and （52.0±6.0）% respectively, which was significantly declined comparing with control group （P 〈 0.05）. The morphological of tumor cells in control group remain normal size and grow eugenic, while the cells of DM group, US group and DM ＋ US group decreased, their shapes were irregular, the intercellular space increased, and cells apoptosis occured in the medium. The cells in DM ＋ US group were changed significantly. FCM showed the apoptosis rate of DM ＋ US group was （26.81 ± 2.82）% ,which was declined clearly compared with （2.90 ± 0.99）% of the control group （ P〈0.05）. Western-blot showed that the expression of proliferating cell nuclear antigen and p-ERK1/2 protein in DM ＋ US group were down-regulated, and meanwhile p-p38 protein in human liver cancer cells MHCC97- H was up-regulated. Conclusions The method of ultrasound irradiating docetaxel-loaded microbubbles can significantly induce human liver cancer cells MHCC97-H apoptosis by regulating the expression of related protein.