Lang Chunhui, Xu Miao, Zhao Ziyi, Chen Jinyao, Zhang Lishi
Department of Nutrition, Food Safety and Toxicology, West China School of Public Health, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Department of Clinical Nutrition, Chongqing Three Gorges Central Hospital, Wanzhou, Chongqing 404000, P.R. China.
Oncol Lett. 2018 Sep;16(3):4020-4026. doi: 10.3892/ol.2018.9122. Epub 2018 Jul 11.
MicroRNA-96 (miR-96) is transcriptionally associated with the induction of chemoresistance following chemotherapy by targeting to FOXO1 mRNA at one of two predicted binding sites in its 3'-untranslated region sequence. The upregulation of miR-96 is associated with a high risk of chemoresistance. Nevertheless, the mechanism by which miR-96 is upregulated remains largely undefined. In the present study, the gastric cancer SGC7901 cell line was treated with different doses of the chemotherapeutic agents cisplatin and doxorubicin. miR-96 expression was analyzed by reverse transcription-quantitative polymerase chain reaction at different time points. Western blot and chromatin immunoprecipitation were performed to analyze the expression levels of the target gene. The effects of miR-96 on chemosensitivity were assessed by a carboxyfluorescein succinimidyl ester/propidium iodide labeling assay, and its effects on proliferation were assessed by Cell Counting Kit-8 or EdU staining assays. The results demonstrated that treatment with a low dose of either chemotherapeutic agent induced miR-96 expression. Upregulation of miR-96 caused the post-transcriptional repression of FOXO1 expression. Decreases in FOXO1 protein levels led to a decrease in the transcriptional activity of the cyclin-dependent kinase inhibitor 1A (CDKN1A, also known as p21) promoter region, and thus the expression of p21 was downregulated in a tumor protein p53-independent manner. As a result, induction of miR-96 expression caused chemoresistance and promoted proliferation in SGC7901 cells. Taken together, the results of the present study revealed that treatment with cisplatin or doxorubicin could induce expression of miR-96 at certain doses. Upregulation of miR-96 is partially associated with chemoresistance and miR-96 can also promote cell proliferation by repressing p21.
微小RNA-96(miR-96)在化疗后通过靶向其3'-非翻译区序列中两个预测结合位点之一的FOXO1 mRNA,在转录水平上与化疗耐药性的诱导相关。miR-96的上调与化疗耐药的高风险相关。然而,miR-96上调的机制在很大程度上仍不明确。在本研究中,胃癌SGC7901细胞系用不同剂量的化疗药物顺铂和阿霉素处理。在不同时间点通过逆转录-定量聚合酶链反应分析miR-96表达。进行蛋白质免疫印迹和染色质免疫沉淀以分析靶基因的表达水平。通过羧基荧光素琥珀酰亚胺酯/碘化丙啶标记试验评估miR-96对化疗敏感性的影响,通过细胞计数试剂盒-8或EdU染色试验评估其对增殖的影响。结果表明,用低剂量的任何一种化疗药物处理均可诱导miR-96表达。miR-96的上调导致FOXO1表达的转录后抑制。FOXO1蛋白水平的降低导致细胞周期蛋白依赖性激酶抑制剂1A(CDKN1A,也称为p21)启动子区域的转录活性降低,因此p21的表达以不依赖肿瘤蛋白p53的方式下调。结果,miR-96表达的诱导导致SGC7901细胞的化疗耐药并促进增殖。综上所述,本研究结果表明,顺铂或阿霉素处理在一定剂量下可诱导miR-96表达。miR-96的上调部分与化疗耐药相关,并且miR-96还可通过抑制p21促进细胞增殖。