二硫键异构酶TMX1促进肺癌化疗耐药

doi:10.6040/j.issn.1671-7554.0.2023.0169

摘要/Abstract

摘要： 目的探讨内质网二硫键异构酶(PDI)在肺癌发生及化疗药物治疗中的作用。方法利用Human Protein Atlas软件数据库筛选在肺组织有表达的PDI家族成员;利用qRT-PCR技术对临床10例患者的肺癌及癌旁组织中的部分PDI家族成员mRNA水平差异进行分析,进一步利用免疫印迹技术对临床4~6例患者的肺癌及癌旁组织部分PDI家族成员的蛋白表达差异进行分析,并利用免疫组化技术进行验证,分析变化显著的差异基因与患者预后的关系,重点考察TMX1在肺癌发生发展中的作用。利用qRT-PCR、免疫印迹技术分析TMX1在肺癌细胞、肺癌多药耐药细胞及非肿瘤上皮细胞的转录和蛋白水平。通过脂质体转染构建过表达和敲低TMX1的细胞,利用MTT实验检测TMX1对肺癌细胞存活的影响,并分析TMX1对化疗药物顺铂和多西紫杉醇药效的影响。结果临床样本检测结果显示,与癌旁组织相比,肺癌组织中PDIA2的mRNA和蛋白水平降低,PDIA3、PDIA6、TMX1、TMX3的mRNA水平均明显升高。对TCGA数据库基因表达和预后关系分析结果显示,PDIA2、TMX1低表达者预后好;而PDIA6高表达时生存期略有延长,PDIA3、TMX3的表达高,生存期显著延长。对TMX1的功能进行基础表达分析,结果显示,相对于非肿瘤的上皮细胞,TMX1在不同类型肺癌细胞的转录水平都较高,但其蛋白表达在肺腺癌A549细胞、小细胞肺癌H446细胞中明显升高;发现TMX1在肺癌的耐药细胞中表达升高,敏感细胞经顺铂和多西紫杉醇处理后其表达上调。下调TMX1的表达,肺癌A549细胞存活减少,且对顺铂和多西紫杉醇的敏感性增加;而在低表达的H1688细胞中过表达TMX1,细胞的存活增加,对顺铂和多西紫杉醇更加耐受。结论 TMX1能够促进肺癌细胞的存活并对顺铂和多西紫杉醇耐药。

关键词: 肺癌, TMX1, 顺铂耐药, 多西紫杉醇耐药, 蛋白质二硫键异构酶

Abstract: Objective To investigate the role of protein disulfide isomerase(PDI)in lung cancer development and chemotherapy treatment. Methods PDI family members expressed in lung tissue were screened using the Human Protein Atlas. The mRNA levels of some PDI family members in lung cancer and adjacent tissues of 10 patients were analyzed with qRT-PCR. The protein expressions of some PDI family members in cancer tissues and adjacent tissues of 4 to 6 patients were determined with Western blotting and verified with immunohistochemistry, in order to explore the relationship between the significantly changed differential genes and the prognosis of patients, especially the role of TMX1 in the occurrence and development of lung cancer. The transcription and protein levels of TMX1 in lung cancer cells, lung cancer multi-drug resistant cells and non-tumor epithelial cells were detected with qRT-PCR and Western blotting. After cells with overexpression and knockdown of TMX1 were constructed with liposome transfection, the effects of TMX1 on the survival of lung cancer cells were detected with MTT assay, and the effects of TMX1 on the efficacy of cisplatin and docetaxel were analyzed. Results Compared with the adjacent tissues, cancer tissues exhibited significantly reduced mRNA and protein levels of PDIA2, significantly increased mRNA levels of PDIA3, PDIA6, TMX1 and TMX3. Analysis of TCGA database showed that lower expressions of PDIA2 and TMX1 were associated with better prognosis, higher expression of PDIA6 was associated with slightly prolonged survival, and higher expressions of PDIA3 and TMX3 were associated with significantly prolonged survival. The transcription level of TMX1 was higher in different types of lung cancer cells than in non-tumor epithelial cells; its protein expression was significantly increased in lung adenocarcinoma A549 cells and small cell lung cancer H446 cells. TMX1 was highly expressed in drug-resistant cells of lung cancer, and its expression in sensitive cells was upregulated after cisplatin and docetaxel treatment. After knockdown of TMX1, the survival of lung cancer A549 cells was reduced and the sensitivity to cisplatin and docetaxel was increased. Overexpression of TMX1 in low-expression H1688 cells resulted in increased cell survival and greater tolerance to cisplatin and docetaxel. Conclusion TMX1 promotes survival of lung cancer cells and resistance to cisplatin and docetaxel.

Key words: Lung cancer, TMX1, Cisplatin resistance, Docetaxel resistance, Protein disulfide isomerase

中图分类号:

R574

王亚楠,梁传杰,贾梦琪,苑辉卿. 二硫键异构酶TMX1促进肺癌化疗耐药[J]. 山东大学学报 (医学版), 2023, 61(8): 31-39.

WANG Yanan, LIANG Chuanjie, JIA Mengqi, YUAN Huiqing. Role of protein disulfide isomerase TMX1 in promoting chemotherapy resistance in lung cancer[J]. Journal of Shandong University (Health Sciences), 2023, 61(8): 31-39.

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