热疗对喉鳞癌患者细胞凋亡的影响及机制研究

Effect and mechanism of hyperthermia on cell apoptosis of laryngeal squamous cell carcinoma patients

目的探讨热疗对喉鳞癌细胞凋亡和胱天蛋白酶3(Caspase-3)及磷酸化的蛋白激酶B(p-AKT)表达的影响。方法对2021年10月至2022年10月于郑州大学第一附属医院就诊的30例喉鳞癌患者进行前瞻性研究, 行3次热疗, 每次间隔24 h。采集30例患者热疗前后的肿瘤组织标本, 分为热疗前组(A组)和热疗后组(B组), 采用自身对照研究进行对比分析, 以TUNEL法检测细胞凋亡情况, 免疫组化检测组织中Caspase-3及p-AKT的表达水平, 分别记录阳性细胞比率及组织化学评分, 两组间的比较采用配对t检验。凋亡程度与Caspase-3及p-AKT分子变化的关联性分析采用Pearson相关分析。结果 30例患者热疗后均无明显不良反应。A组喉鳞癌细胞的凋亡指数为2.37%±1.33%, B组为4.27%±3.93%(P=0.006)。A组肿瘤组织中Caspase-3的阳性细胞比率为62.31%±19.49%, B组为80.79%±17.15%(P=0.001);A组p-AKT阳性细胞比率为31.26%±19.30%, B组为26.26%±15.86%(P=0.023)。凋亡程度的变化与Caspase-3分子的变化之间存在正相关关系(r=0.544, P=0.002), 而与p-AKT分子的变化呈负相关关系(r=-0.434, P=0.017)。结论热疗可以促进喉鳞癌肿瘤组织的细胞凋亡, 可能与Caspase-3依赖性凋亡有关, 而AKT磷酸化受抑制也参与了这一过程。

Objective To evaluate the effect of hyperthermia on the apoptosis and the expression levels of cysteine-containing aspartate-specific protease-3(Caspase-3)and phosphorylated protein kinase B(p-AKT)in laryngeal squamous cell carcinoma cells.Methods A prospective study was conducted on 30 patients with laryngeal squamous cell carcinoma who were treated at the First Affiliated Hospital of Zhengzhou University from October,2021 to October,2022.Three times of hyperthermia were performed with a time interval of 24 h.The tumor tissue samples were collected from 30 patients before and after hyperthermia and divided into before hyperthermia group(group A)and after hyperthermia group(group B).Self-control study mode was adopted for comrparative analysis.The cell apoptosis was detected by TUNEL assay.The expression levels of Caspase-3 and p-AKT in the tissues were detected by immunohistochemistry.Positive cell ratio and immunohistochemistry(IHC)score were recorded.Comparison between two groups was performed by paired t-test.The correlation between the degree of apoptosis and the changes of Caspase-3 and p-AKT molecules was assessed by Pearson correlation analysis.Results No evident adverse reactions were observed in 30 patients after hyperthermia.The apoptosis index of laryngeal squamous cell carcinoma cells in group A was 2.37%±1.33%,and 4.27%±3.93%in group B(P=0.006).In group A,the ratio of Caspase-3 positive cells in tumor tissues was 62.31%±19.49%and 80.79%±17.15%in group B(P=0.001).The ratio of p-AKT positive cells in group A was 31.26%±19.30%,and 26.26%±15.86%in group B(P=0.023).There was a positive correlation between the degree of apoptosis and the changes of Caspase-3 molecule(r=0.544,P=0.002),but a negative correlation was noted between the degree of apoptosis and the changes of p-AKT molecule(r=-0.434,P=0.017).Conclusion Hyperthermia can promote the apoptosis of tumor cells in laryngeal squamous cell carcinoma,which may be related to Caspase-3 dependent apoptosis,and the inhibition of AKT phosphorylation is also involved in this process.