液态氟碳纳米粒液-气相变治疗肝癌的体外实验

Phase transition of liquid perfluorocarbon nanoparticles in treatment of hepatocellular carcinoma in vitro

目的探讨低强度聚焦超声(LIFU)辐照包裹液态氟碳(PFP)脂质纳米粒(L-NP)致液-气相变(ADV)对肝癌HepG2细胞凋亡和存活的影响。方法制备包裹PFP的L-NP。在加热条件下光镜观察包裹PFP的L-NP相变情况,超声观察LIFU辐照后包裹PFP的L-NP相变情况,荧光显微镜观察人肝癌细胞吞噬L-NP情况,LIFU辐照靶向组、非靶向组、对照组肝癌细胞后应用流式细胞术检测肝癌细胞凋亡情况,CCK-8检测肝癌细胞存活情况。并进行统计学分析。结果包裹PFP的L-NP在44.5℃时开始出现ADV,随温度升高相变纳米粒由少到多,由小到大,体积达到一定程度时瞬间爆破。包裹PFP的L-NP经LIFU辐照后声像图回声较辐照前增强。靶向L-NP能特异性聚集到肝癌细胞周围并被肝癌细胞吞噬。LIFU辐照后,靶向组肝癌细胞凋亡率[(95.86±1.96)%]高于非靶向组[(17.40±2.20)%]和对照组[(7.70±1.19)%],存活率[(49.21±3.04)%]低于非靶向组[(84.24±2.70)%],差异均有统计学意义(P均<0.05)。结论 LIFU辐照可致靶向到肝癌细胞的包裹PFP的L-NP发生ADV,从而促使其死亡,抑制肝癌细胞增殖。

Objective To explore the effect of apoptosis and survival of the hepatocellar carcinoma HepG2 cells encountered phase transition of liquid fluorocarbon（PFP）lipid nanoparticals（L-NP）which were caused by low intensity focused ultrasound（LIFU）irradition.Methods L-NP was prepared.The phase transition of L-NP was observed using light microscope under the condition of heating and using ultrasound diagnostic apparatus after LIFU irradiation.The phenomenon of the human hepatocellar carcinoma cells swallowing L-NP was observed using inverted flourescence microscope.The apoptosis of hepatoma cells in target group,non-target group and control group after LIFU irradiation were detected using flow cytometry detection.And the survival of hepatoma cells were detected using CCK-8 reagent.Moreover,the statistical analysis was preformed.Results Under the Condition of 44.5℃,L-NP with PFP inside began to occur ADV.The quantity and volume of shifted L-NP were increased with the temperature.It wound burst instantaneously when the volume achieved threshold.The intensity in sonogram of L-NP with PFP after LIFU irradiation was significantly higher than before.The targeted L-NP could gathered specially around of hepatoma cells and be swallowed by them,After LIFU irradiation,the apoptosis rate of hepatoma cells in target group（[95.86±1.96]%）was respectively higher than that in non-target group（[17.40±2.20]%）and control group（[7.70±1.19]%,both P〈0.05）.And the survival rate（[49.21±3.04]%）was lower than that in non-target group（[84.24±2.70]%,P〈0.05）.Conclusion The targeted L-NP occured ADV caused by LIFU irradiation which can kill and inhibit hepatocellular carcinoma cells.