Institute of Clinical Pharmacology, Hunan Key Laboratory of Pharmacogenetics, Xiang-Ya Hospital, Central South University, Changsha, Hunan, PR China.
Clin Cancer Res. 2011 Jul 1;17(13):4600-9. doi: 10.1158/1078-0432.CCR-10-2591. Epub 2011 May 24.
The purpose of this study is to test the hypothesis that eIF3a may regulate the expression of DNA repair proteins which, in turn, affects response of lung cancer patients to treatments by DNA-damaging anticancer drugs.
Immunohistochemistry was used to determine the expression of eIF3a in 211 human lung cancer tissues followed by association analysis of eIF3a expression with patient's response to platinum-based chemotherapy. Ectopic overexpression and RNA interference knockdown of eIF3a were carried out in NIH3T3 and H1299 cell lines, respectively, to determine the effect of altered eIF3a expression on cellular response to cisplatin, doxorubicine, etoposide (VP-16), vincristine, and vinblastine by using MTT assay. The DNA repair capacity of these cells was evaluated by using host-cell reactivation assay. Real-time reverse transcriptase PCR and Western Blot analyses were carried out to determine the effect of eIF3a on the DNA repair genes by using cells with altered eIF3a expression.
eIF3a expression associates with response of lung cancer patients to platinum-based chemotherapy. eIF3a knockdown or overexpression, respectively, increased and decreased the cellular resistance to cisplatin and anthrocycline anticancer drugs, DNA repair activity, and expression of DNA repair proteins.
eIF3a plays an important role in regulating the expression of DNA repair proteins which, in turn, contributes to cellular response to DNA-damaging anticancer drugs and patients' response to platinum-based chemotherapy.
本研究旨在验证假设，即 eIF3a 可能调节 DNA 修复蛋白的表达，而 DNA 修复蛋白的表达反过来又会影响肺癌患者对 DNA 损伤抗癌药物治疗的反应。
通过免疫组织化学检测 211 例人肺癌组织中 eIF3a 的表达，然后将 eIF3a 表达与患者对铂类化疗的反应进行关联分析。分别在 NIH3T3 和 H1299 细胞系中进行 eIF3a 的异位过表达和 RNA 干扰敲低，通过 MTT 测定法确定改变 eIF3a 表达对顺铂、阿霉素、依托泊苷（VP-16）、长春新碱和长春碱的细胞反应的影响。通过宿主细胞复活测定评估这些细胞的 DNA 修复能力。通过改变 eIF3a 表达的细胞进行实时逆转录 PCR 和 Western Blot 分析，确定 eIF3a 对 DNA 修复基因的影响。
eIF3a 的表达与肺癌患者对铂类化疗的反应相关。eIF3a 敲低或过表达分别增加和降低了细胞对顺铂和蒽环类抗癌药物、DNA 修复活性以及 DNA 修复蛋白表达的耐药性。
eIF3a 在调节 DNA 修复蛋白的表达中起重要作用，而 DNA 修复蛋白的表达又有助于细胞对 DNA 损伤抗癌药物的反应和患者对铂类化疗的反应。
