低频超声联合微泡剂诱导肝癌细胞凋亡的实验研究

摘要

Objective To investigate the affect of NF-κB through low-frequency ultrasound combined with microbubble agent inducing the apoptosis of human hepatocellular carcinoma SMMC-7721 cells.Methods Cells were divided into four groups.They were blank group ( A), microbubble agent group ( B), low-frequency ultrasound group(C) and low frequency ultrasound combined with microbubble agent group(D).Cell apoptosis was analyzed by flow cytometry.Transmission electron microscope was used to observe cell morphous; Western blotting was used to analyze the expression of NF-κB.Results The apoptosis rate of the 4 group were ( 1.51 ± 0.21 ) ％, (2.49 ± 0.30) ％, ( 14.14 ± 2.01 ) ％ and(27.01 ± 4.21 ) ％ respectively.Differences were not obvious between group A and B.The apoptosis rates of group C and D were higher than that of group A ( P ＜0.01 ), and the apoptosis rate of group D was higher than that of group C ( P ＜ 0.05 ).Morphology alterations of group D were obviously observed using transmission electron microscope.NF-κB expression in group D was lower than other groups, and the grey intensity ratios of A-D groups were 5.34, 4.29, 2.72 and 1.04 respectively.Conclusion Low-frequency ultrasound combined with microbubble agent can induce apoptosis of human hepatocellular carcinoma cells by decreasing the expression of NF-κB, and microbubble agent can strengthen the effect of low frequency ultrasound.%目的 研究核因子-κB(NF-κB)在低频超声联合微泡剂对肝癌SMMC-7721细胞诱导凋亡中的作用.方法 选取肝癌细胞SMMC-7721,分为空白对照组(A)、微泡剂组(B)、低频超声组(C)和低频超声联合微泡剂组(D).流式细胞术检测肝癌细胞凋亡情况,透射电镜观察细胞形态改变,Western blotting检测癌细胞NF-κB的表达.结果 A、B、C、D组肝癌细胞的凋亡率分别为(1.51±0.21)%、(2.49±0.30)%、(14.14±2.01)%和(27.01±4.21)%.C、D两组均明显高于A组(P＜0.01),D组亦高于C组(P＜0.05).透射电镜观察D组细胞呈凋亡形态学改变.经Western blotting检测,D组NF-κB蛋白表达明显低于其他各组,A～D组NF-κB灰度值比值分别为5.34、4.29、2.72和1.04.结论 低频超声联合微泡剂通过抑制NF-κB的表达促进肝癌细胞的凋亡,其中微泡剂可增强低频超声的凋亡效应.