Experimental study on ultrasound-guided radiofrequency of hyperthermia combined with recombinant human adenovirus H101 for the treatment of hepatocellular carcinoma / 中华肝胆外科杂志

ABSTRACT

Objective:Evaluation of ultrasound-guided radiofrequency of hyperthermia combined with recombinant human adenovirus H101 for the treatment of hepatocellular carcinoma (HCC).Methods:In vitro cell therapy experiments, luciferase/red fluorescent protein/lentivirus mediated McA-RH7777 cells were conducted and divided into 4 groups. Each group was repeatedly treated for 6 times (1) recombinant human adenovirus type 5 H101 [(multiplicity of infection, MOI)=0.2]+ RFH group, heated at 42℃ for 30 min; (2) recombinant human adenovirus type 5 H101 alone (MOI=0.2); (3) RFH alone, heated at 42℃ for 30 min; (4) control group physiological saline group. Twenty-four nude rats weighing 180-220 g were selected to establish a nude rat model of orthotopic HCC. They were divided into 4 groups with 6 rats in each group (1) H101+ RFH combined treatment group RFH electrode needles were punctured to the center of the tumor in the liver of nude rats under ultrasound guidance, and H101 was directly injected through the electrode injection end. RFH was delivered to the tumor at a temperature of 42℃ for 30 min; (2) H101 treatment group MOI=0.2; (3) RFH treatment group; (4) sham surgery group. Fluorescence microscopy imaging was used to evaluate the viability of cells in vitro experiments. For in vivo validation, ultrasound imaging was used to follow up the tumor size. Tumor gross specimens and pathological changes were also evaluated.Results:Twenty-four hours after treatment, the survival rate of cells in the H101+ RFH group was the lowest under fluorescence microscopy. The results of MTS quantitative analysis showed that the relative absorbance of mezzanine in the H101+ RFH group cells was lower than that in the H101 group alone [(25.00±2.27)% vs. (69.50±4.53)%], the RFH group alone [(25.00±2.27)% vs. (92.83±1.66)%], and the control group [(25.00±2.27)% vs. 100%], with statistical significance (all P<0.001). The number of apoptotic cells in the H101+ RFH group was higher than that in the H101 group alone [(54.5±3.1)% vs. (25.2±1.4)%], the RFH group alone [(54.5±3.1)% vs. (5.7±0.6)%], and the control group [(54.5±3.1)% vs. (3.9±0.5)%], all of which showed statistically significant differences (all P<0.001). The relative tumor volume of nude rats in the H101+ RFH combination treatment group was smaller than that in the H101 treatment group (0.776±0.127 vs. 1.312±0.188), RFH treatment group (0.776±0.127 vs. 1.893±0.571), and sham surgery group (0.776±0.127 vs. 1.977±0.590), all of which had statistical significance (all P<0.001). The number of apoptotic cells in nude rats in the H101+ RFH combination treatment group was higher than that in the H101 treatment group [(49.85±4.00%)% vs. (22.70±0.65)%], the RFH treatment group [(49.85±4.00% vs. (5.36±0.84)%], and the sham surgery group [(49.85±4.00)% vs. (5.96±0.78)%], all of which showed statistically significant differences (all P<0.001). Conclusion:Ultrasound guided RFH combined with recombinant human adenovirus H101 has a promoting effect on the treatment of HCC.