Department of Otorhinolaryngology, Head and Neck Surgery, The First Clinical Hospital, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Braz J Med Biol Res. 2013 Jun;46(6):546-54. doi: 10.1590/1414-431X20131662. Epub 2013 Jun 12.
Multidrug resistance (MDR) poses a serious impediment to the success of chemotherapy for laryngeal cancer. To identify microRNAs and mRNAs associated with MDR of human laryngeal cancer Hep-2 cells, we developed a multidrug-resistant human laryngeal cancer subline, designated Hep-2/v, by exposing Hep-2 cells to stepwise increasing concentrations of vincristine (0.02-0.96'µM). Microarray assays were performed to compare the microRNA and mRNA expression profiles of Hep-2 and Hep-2/v cells. Compared to Hep-2 cells, Hep-2/v cells were more resistant to chemotherapy drugs (≈ 45-fold more resistant to vincristine, 5.1-fold more resistant to cisplatin, and 5.6-fold more resistant to 5-fluorouracil) and had a longer doubling time (42.33 ± 1.76 vs 28.75 ± 1.12'h, P<0.05), higher percentage of cells in G0/G1 phase (80.98 ± 0.52 vs 69.14 ± 0.89, P<0.05), increased efflux of rhodamine 123 (95.97 ± 0.56 vs 12.40 ± 0.44%, P<0.01), and up-regulated MDR1 expression. A total of 7 microRNAs and 605 mRNAs were differentially expressed between the two cell types. Of the differentially expressed mRNAs identified, regulator of G-protein signaling 10, high-temperature requirement protein A1, and nuclear protein 1 were found to be the putative targets of the differentially expressed microRNAs identified. These findings may open a new avenue for clarifying the mechanisms responsible for MDR in laryngeal cancer.
多药耐药（MDR）严重阻碍了喉癌化疗的成功。为了鉴定与人类喉癌细胞 Hep-2 的 MDR 相关的 microRNAs 和 mRNAs，我们通过逐步增加长春新碱（0.02-0.96'µM）的浓度来暴露 Hep-2 细胞，从而开发了多药耐药的人类喉癌细胞亚系，命名为 Hep-2/v。通过 microarray 分析比较了 Hep-2 和 Hep-2/v 细胞的 microRNA 和 mRNA 表达谱。与 Hep-2 细胞相比，Hep-2/v 细胞对化疗药物的耐药性更强（对长春新碱的耐药性约为 45 倍，对顺铂的耐药性为 5.1 倍，对 5-氟尿嘧啶的耐药性为 5.6 倍），倍增时间更长（42.33 ± 1.76 vs 28.75 ± 1.12'h，P<0.05），G0/G1 期细胞比例更高（80.98 ± 0.52 vs 69.14 ± 0.89，P<0.05），罗丹明 123 外排增加（95.97 ± 0.56 vs 12.40 ± 0.44%，P<0.01），并且 MDR1 表达上调。两种细胞类型之间共表达了 7 个 microRNAs 和 605 个 mRNAs。在鉴定出的差异表达的 mRNAs 中，G 蛋白信号调节因子 10、热休克蛋白 A1 和核蛋白 1 被鉴定为差异表达 microRNAs 的潜在靶标。这些发现可能为阐明喉癌 MDR 的机制开辟新途径。
