2023 年第 10 期 第 18 卷

肿瘤耐药相关基因对骨肉瘤新辅助化疗耐药的评估价值

Evaluation of tumor resistance related genes in neoadjuvant chemotherapy resistance for osteosarcoma

作者：李斌1李战鹏1连振刚1刘亚云2

英文作者：Li Bin1 Li Zhanpeng1 Lian Zhengang1 Liu Yayun2

单位：1河北省张家口市第一医院骨科，张家口075000；2江西省人民医院骨科，南昌330006

英文单位：1Department of Orthopedics Zhangjiakou First Hospital Hebei Province Zhangjiakou 075000 China; 2Department of Orthopedics Jiangxi Provincial People′s Hospital Nanchang 330006 China

关键词：骨肉瘤；第10号染色体缺失性磷酸酶-张力蛋白同源物基因；DNA拓扑异构酶Ⅱ；多药耐药基因1；新辅助化疗耐药

英文关键词：Osteosarcoma;Phosphataseandtensinhomologydeletedonchromosometen;DNAtopoisomeraseⅡ;Multidrugresistancegene-1;Neoadjuvantchemotherapyresistance

• 摘要：

• 目的  探究第10号染色体缺失性磷酸酶-张力蛋白同源物基因（PTEN）、DNA拓扑异构酶Ⅱ（TOPOⅡ）、多药耐药基因1（MDR1）对骨肉瘤新辅助化疗耐药的评估价值。方法  选取2020年7月至2022年7月在河北省张家口市第一医院确诊并治疗的106例骨肉瘤患者，根据新辅助化疗结果分为化疗敏感组（74例）和化疗耐药组（32例）。采用免疫组织化学染色法、蛋白质印迹法、实时荧光定量聚合酶链反应（RT-PCR）法检测PTEN、TOPOⅡ、MDR1蛋白表达情况。采用Logistic回归模型分析骨肉瘤新辅助化疗耐药的影响因素。采用受试者工作特征曲线分析外周血PTEN、TOPOⅡ、MDR1 mRNA表达水平对骨肉瘤新辅助化疗耐药的评估价值。结果  免疫组织化学染色结果显示，化疗耐药组PTEN阳性表达率低于化疗敏感组(P＜0.05)，TOPOⅡ、MDR1阳性表达率均高于化疗敏感组(均P＜0.05)。蛋白质印迹法结果显示，化疗耐药组PTEN表达量低于化疗敏感组，TOPOⅡ和MDR1表达量均高于化疗敏感组(均P＜0.05)。RT-PCR法结果显示，化疗耐药组PTEN mRNA表达水平低于化疗敏感组［（0.34±0.07）比（2.87±0.69）］，TOPOⅡ、MDR1 mRNA表达水平均高于化疗敏感组［（3.22±0.77）比（0.86±0.21），（3.00±0.67）比（0.94±0.23）］(均P＜0.05)。结论  PTEN、TOPOⅡ、MDR1联合检测对骨肉瘤新辅助化疗耐药有较好的评估作用，这对于早期筛选出新辅助化疗耐药患者有重要的指导意义。

• Objective  To investigate the evaluation of phosphatase and tensin homology deleted on chromosome ten (PTEN), DNA topoisomerase Ⅱ (TOPOⅡ) and multidrug resistance gene-1 (MDR1) in neoadjuvant chemotherapy resistance for osteosarcoma. Methods  From July 2020 to July 2022, 106 patients with osteosarcoma admitted to Zhangjiakou First Hospital, Hebei Province were collected. They were divided into chemotherapy sensitive group (74 cases) and chemotherapy resistant group (32 cases) according to the results   of neoadjuvant chemotherapy. Immunohistochemical staining, western blotting and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of PTEN, TOPOⅡ and MDR1 proteins. Logistic regression model was used to analyze the influencing factors of neoadjuvant chemotherapy resistance for osteosarcoma. Receiver operating characteristic curve was used to analyze the expression levels of PTEN, TOPOⅡ and MDR1 mRNA of peripheral blood in neoadjuvant chemotherapy resistance for osteosarcoma. Results  The results   of immunohistochemical staining showed that the positive expression rate of PTEN in the chemotherapy resistant group was lower than that in the chemotherapy sensitive group (P<0.05), and the positive expression rates of TOPOⅡ and MDR1 were both higher than those in the chemotherapy sensitive group (both P<0.05). The results   of western blotting showed that the expression of PTEN in the chemotherapy resistant group was lower than that in the chemotherapy sensitive group, and the expressions of TOPO Ⅱ and MDR1 in the chemotherapy resistant group were higher than those in the chemotherapy sensitive group (all P<0.05). The results   of RT-PCR showed that the expression of PTEN mRNA of peripheral blood in the chemotherapy resistant group was lower than that in the chemotherapy sensitive group［(0.34±0.07) vs (2.87±0.69)］, and the expressions of TOPOⅡ and MDR1 mRNA in the chemotherapy resistant group were both higher than those in the chemotherapy sensitive group ［(3.22±0.77) vs (0.86±0.21), (3.00±0.67) vs (0.94±0.23)］(all P<0.05). Conclusion  The combined detection of PTEN, TOPOⅡ and MDR1 has a good evaluation effect in neoadjuvant chemotherapy resistance for osteosarcoma, and this has an important guiding significance for the early screening of patients with neoadjuvant chemotherapy resistance.