超声引导下射频消融治疗肝癌的体外研究

目的探索超声引导下射频消融治疗肝癌的有效消融模式。方法观察人肝癌细胞HepG2和人永久化肝细胞LO2经高温处理后的形态变化,采用细胞计数试剂盒8(CCK-8)法检测其增殖能力。收集47℃处理后仍然存活的HepG2细胞(ΔHepG2细胞)和LO2细胞(ΔLO2细胞),行二次高温处理,采用CCK-8法检测其增殖能力。在离体猪肝的不同位置进行消融,比较超声声像图汽化区与肉眼观消融灶的一致性,观察声像图汽化区各区域的温度。结果水浴加热至50℃后,随着培养时间的延长,HepG2和LO2细胞的贴壁细胞数目愈来愈少,且逐渐失去正常细胞形态。水浴加热至50℃及以上后,HepG2、LO2、ΔHepG2和ΔLO2细胞的细胞增殖率降低,与37℃组比较,差异均有统计学意义(均P<0.05),并对细胞产生不可逆性损伤。声像图中汽化区横径与肉眼观消融灶横径较为一致,差异均无统计学意义(均P>0.05)。射频消融9 min时,超声图像上高回声团边缘及其内部猪肝组织温度均≥53℃,为有效消融时间。结论初始功率200 W、目标温度105℃,当有效消融时间持续9 min以上时,超声声像图汽化区内的肝癌细胞均可发生不可逆性损伤。

Ex-vivo experimental study of ultrasound-guided radiofrequency ablation for the treatment of hepatocellular carcinoma

Objective To explore the effective mode of ultrasound-guided radiofrequency ablation for the treatment of hepatocellular carcinoma(HCC).Methods The morphological changes of human liver cancer cells HepG2 and human permanent liver cells LO2 after high temperature treatment were observed.The cell counting kit 8(CCK-8)array was used to detect cell proliferation ability.The HepG2 cells(ΔHepG2 cells)and LO2 cells(ΔLO2 cells)those were still alive after treatment with 47℃were collected and subjected to secondary high temperature treatment,and their proliferation ability was tested by the CCK-8 array.The ablation was performed at different locations of the ex-vivo porcine liver.The consistency of the vaporization area of the ultrasound image with that of the actual ablation was compared,the temperature of each area in the ultrasonic image was observed.Results After incubated water was heated to 50℃,with the extension of the culture time,the number of adherent cells of HepG2 and LO2 cells became less and less,and gradually lost their normal cell morphology.After 50℃and above,all of the proliferation rates of HepG2 cells,LO2 cells,ΔHepG2 cells andΔLO2 cells were decreased.Compared with the 37℃group,the differences were statistically significant(P<0.05),and the cells were irreversible damaged.There was no significant difference between the transverse diameters of the sonographic vaporization zones and the actual ablation zones(P>0.05).Nine minutes was considered to be an effective ablation time point since the temperatures in the hyperechoic areas were all above 53℃.Conclusion The HCC cells within the vaporization zone observed by ultrasound can be damaged irreversibly when they were effectively ablated for over 9 minutes at 105℃and 200 W.