Department of Infectious Diseases, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Gene Hospital of Henan Province, Precision Medicine Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
PLoS One. 2023 Mar 15;18(3):e0273754. doi: 10.1371/journal.pone.0273754. eCollection 2023.
Liver tumor remains an important cause of cancer-related death. Nanosecond pulsed electric fields (nsPEFs) are advantageous in the treatment of melanoma and pancreatic cancer, but their therapeutic application on liver tumors need to be further studied.
Hep3B cells were treated with nsPEFs. The biological behaviors of cells were detected by Cell Counting Kit-8, 5-ethynyl-20-deoxyuridine, and transmission electron microscopy (TEM) assays. In vivo, rabbit VX2 liver tumor models were ablated by ultrasound-guided nsPEFs and radiofrequency ablation (RFA). Contrast-enhanced ultrasound (CEUS) was used to evaluate the ablation effect. HE staining and Masson staining were used to evaluate the tissue morphology after ablation. Immunohistochemistry was performed to determine the expression of Ki67, proliferating cell nuclear antigen, and α-smooth muscle actin at different time points after ablation.
The cell viability of Hep3B cells was continuously lower than that of the control group within 3 days after pulse treatment. The proliferation of Hep3B cells was significantly affected by nsPEFs. TEM showed that Hep3B cells underwent significant morphological changes after pulse treatment. In vivo, CEUS imaging showed that nsPEFs could completely ablate model rabbit VX2 liver tumors. After nsPEFs ablation, the area of tumor fibrosis and the expression of Ki67, proliferating cell nuclear antigen, and α-smooth muscle actin were decreased. However, after RFA, rabbit VX2 liver tumor tissue showed complete necrosis, but the expression of PCNA and α-smooth muscle actin did not decrease compared to the tumor group.
nsPEFs can induce Hep3B cells apoptosis and ablate rabbit VX2 liver tumors in a non-thermal manner versus RFA. The ultrasound contrast agent can monitor immediate effect of nsPEF ablation. This study provides a basis for the clinical study of nsPEFs ablation of liver cancer.
肝癌仍然是癌症相关死亡的重要原因。纳秒级脉冲电场（nsPEFs）在治疗黑色素瘤和胰腺癌方面具有优势，但它们在肝肿瘤治疗中的应用需要进一步研究。
用 nsPEFs 处理 Hep3B 细胞。通过细胞计数试剂盒-8、5-乙炔基-20-脱氧尿苷和透射电子显微镜（TEM）检测细胞的生物学行为。在体内，通过超声引导 nsPEFs 和射频消融（RFA）消融兔 VX2 肝癌模型。使用超声造影（CEUS）评估消融效果。HE 染色和 Masson 染色评估消融后组织形态。免疫组化检测消融后不同时间点 Ki67、增殖细胞核抗原和α-平滑肌肌动蛋白的表达。
脉冲处理后 3 天内，Hep3B 细胞的细胞活力持续低于对照组。nsPEFs 显著影响 Hep3B 细胞的增殖。TEM 显示 Hep3B 细胞在脉冲处理后发生明显的形态变化。在体内，CEUS 成像显示 nsPEFs 可完全消融模型兔 VX2 肝癌。nsPEFs 消融后，肿瘤纤维化面积及 Ki67、增殖细胞核抗原和α-平滑肌肌动蛋白表达降低。然而，RFA 后，兔 VX2 肝癌组织显示完全坏死，但与肿瘤组相比，PCNA 和α-平滑肌肌动蛋白的表达并未降低。
与 RFA 相比，nsPEFs 以非热方式诱导 Hep3B 细胞凋亡并消融兔 VX2 肝癌。超声造影剂可监测 nsPEF 消融的即时效果。本研究为 nsPEFs 消融肝癌的临床研究提供了依据。
