热化疗诱导损伤相关分子模式表达增强口腔鳞状细胞癌细胞免疫原性的研究

doi:10.7518/hxkq.2020.05.005

摘要/Abstract

摘要： 目的探讨热疗、化疗及热化疗联合能否诱导口腔鳞状细胞癌细胞表达损伤相关分子模式（DAMPs）增强口腔鳞状细胞癌细胞免疫原性。方法采用CCK-8实验确定平阳霉素（PYM）工作浓度,选择温度39、42及45 ℃作用于口腔鳞状细胞癌CAL27、SCC-15和Tca8113细胞。膜联蛋白V（Annexin V）/碘化丙啶（PI）联合染色、流式细胞术以及酶联免疫吸附试验（ELISA）探究热疗、化疗及两者联合对3株口腔鳞状细胞癌细胞凋亡、钙网蛋白（CRT）膜表达以及高迁徙率族蛋白B1（HMGB1）分泌的影响。应用SPSS 20.0对数据进行统计学分析。结果热疗及化疗均可诱导口腔鳞状细胞癌细胞凋亡,并促进CRT的膜表达及HMGB1的胞外分泌;两者联合应用,细胞凋亡、CRT膜表达及HMGB1分泌均较单纯化疗组升高,差异有统计学意义（P<0.05）。结论热疗、化疗及热化疗联合均可在诱导口腔鳞状细胞癌细胞凋亡的同时,促使CRT在细胞膜表面表达,并促进HMGB1的分泌。热化疗联合应用在凋亡诱导以及诱导DAMPs表达方面效果优于单纯化疗。

关键词: 口腔鳞状细胞癌, 热化疗, 免疫原性, 凋亡, 钙网蛋白, 高迁徙族蛋白B1

Abstract:

Objective To investigate whether hyperthermia, chemotherapy and thermo-chemotherapy could trigger the expression of damage-associated molecular patterns (DAMPs). Methods The optimal working concentration of pingyangmycin (PYM) was detected by CCK-8 assay, and temperatures of 39, 42, and 45 ℃ were applied to the oral squamous cell carcinoma CAL27, SCC-15, and Tca8113 cell lines. The effects of different treatments on the apoptosis, calreticulin (CRT) membrane expression and high-mobility group box 1 (HMGB1) secretion of the cells were detected by using Annexin V/propidium iodide (PI), flow cytometry and enzyme-linked immunosorbent assay (ELISA) assay. SPSS 20.0 software was used for statistical analysis. Results Both hyperthermia and chemotherapy could increase the membrane expression of CRT and the secretion of HMGB1, and furthermore, thermo-chemotherapy group showed significantly increased in apoptosis, CRT membrane expression rate and HMGB1 secretion compared with chemotherapy group, and the difference was statistically significant (P<0.05). Conclusion Hyperthermia, chemotherapy and thermo-chemotherapy could induce oral squamous cell carcinoma cells succumb to death, and at the same time, they can effectively induce the membrane expression of CRT, and promote the secretion of HMGB1. Moreover, thermo-chemotherapy is significantly better than that of chemotherapy alone in the induction of cell apoptosis and DAMPs expression.

Key words: oral squamous cell carcinoma, thermo-chemotherapy, immunogenicity, apoptosis, calreticulin, high-mobility group box 1

中图分类号:

R739.8

孙巧珍, 石凡, 罗丹, 徐婷, 王升志. 热化疗诱导损伤相关分子模式表达增强口腔鳞状细胞癌细胞免疫原性的研究[J]. 华西口腔医学杂志, 2020, 38(5): 502-508.

Sun Qiaozhen, Shi Fan, Luo Dan, Xu Ting, Wang Shengzhi. Thermo-chemotherapy enhances the immunogenicity of oral squamous cell carcinoma cells by inducing the expression of damage-associated molecular patterns[J]. West China Journal of Stomatology, 2020, 38(5): 502-508.

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组别	2.5 μg·mL^-1	5 μg·mL^-1	10 μg·mL^-1	20 μg·mL^-1	40 μg·mL^-1	80 μg·mL^-1
CAL27	50.84±4.81	51.49±2.75	62.99±3.76	73.43±4.35	78.18±2.13	82.76±1.64
SCC-15	48.94±3.59	52.41±1.93	53.00±3.53	58.03±3.84	64.18±2.48	64.46±1.71
Tca8113	38.90±3.08	40.12±4.22	45.23±4.01	47.00±4.78	51.22±3.08	55.65±3.02