The Second Affiliated Hospital of Jiaxing University, Pulmonary and Critical Care Medicine, No. 1518, north Huancheng Road Nanhu District, Jiaxing, Zhejiang Province 314000, PR China.
The Second Affiliated Hospital of Jiaxing University, Pulmonary and Critical Care Medicine, No. 1518, north Huancheng Road Nanhu District, Jiaxing, Zhejiang Province 314000, PR China.
Transpl Immunol. 2021 Dec;69:101478. doi: 10.1016/j.trim.2021.101478. Epub 2021 Oct 2.
Cisplatin (DDP) is used for lung cancer therapy. MicroRNAs, small non-coding RNAs, may contribute to tumorigenesis as well as to drug resistance. We examined regulatory functions of miR-106a-5p in DDP-resistant lung cancer cells.
Differentially expressed miRNAs were provided by Gene Expression Omnibus (GEO) datasets and RT-qPCR examined RNA levels of miR-106a-5p and Jumonji domain-containing protein 6 (JMJD6), an enzyme causing lysine hydroxylation and arginine demethylation. Bindings were determined by luciferase reporter assay. Additionally, the half maximal inhibitory concentration (IC) of DDP was determined through Cell Counting Kit-8 (CCK-8) after treated by DDP (0, 6.25, 12.5, 25, 50 and 75 μM) and apoptosis rates were analyzed using flow cytometry. Besides that, migratory ability and invasiveness were examined by transwell. Western blot was for measuring protein levels of Bcl-2, Bax in apoptosis and E-cadherin, N-cadherin in epithelial-mesenchymal transition (EMT).
The IC value of DDP-resistant A549 (A549/DDP) cells was higher, so were migration, invasion and N-cadherin in EMT while the apoptosis and E-cadherin in EMT were lower versus the parental A549 cells (no DDP resistance). MiR-106a-5p was low expressed in A549/DDP cells while its overexpression caused decreased migration, invasiveness and EMT but promoted apoptosis. JMJD6 was directly targeted and negatively regulated by miR-106a-5p. Inhibited JMJD6 decreased migratory ability, invasion and EMT but improved apoptosis. Moreover, knockdown of miR-106a-5p induced high level of JMJD6, migration, invasiveness and EMT but low apoptosis rates, which were restrained by JMJD6 suppression.
MiR-106a-5p/JMJD6 axis accelerated cell apoptosis and suppressed invasiveness, migration and EMT in A549/DDP cells.
顺铂（DDP）用于肺癌治疗。微小 RNA（miRNA）是一类小的非编码 RNA，可能参与肿瘤发生和耐药。我们研究了 miR-106a-5p 在 DDP 耐药肺癌细胞中的调控功能。
通过基因表达综合数据库（GEO）数据集提供差异表达的 miRNA，并用 RT-qPCR 检测 miR-106a-5p 和 jumonji 结构域包含蛋白 6（JMJD6）的 RNA 水平，JMJD6 是一种导致赖氨酸羟化和精氨酸去甲基化的酶。通过荧光素酶报告基因检测确定结合。此外，用细胞计数试剂盒-8（CCK-8）测定 DDP（0、6.25、12.5、25、50 和 75 μM）处理后 DDP 的半最大抑制浓度（IC），并用流式细胞术分析凋亡率。此外，用 Transwell 检测迁移和侵袭能力。用 Western blot 测定凋亡中 Bcl-2、Bax 和上皮间质转化（EMT）中 E-钙黏蛋白、N-钙黏蛋白的蛋白水平。
DDP 耐药 A549（A549/DDP）细胞的 IC 值较高，迁移、侵袭和 EMT 中的 N-钙黏蛋白较高，而凋亡和 EMT 中的 E-钙黏蛋白较低。miR-106a-5p 在 A549/DDP 细胞中低表达，而过表达导致迁移、侵袭和 EMT 减少，凋亡增加。JMJD6 是 miR-106a-5p 的直接靶标和负调控因子。抑制 JMJD6 减少迁移、侵袭和 EMT，但促进凋亡。此外，miR-106a-5p 敲低诱导 JMJD6 高水平、迁移、侵袭和 EMT 但凋亡率低，抑制 JMJD6 可抑制这些作用。
miR-106a-5p/JMJD6 轴加速 A549/DDP 细胞的细胞凋亡，抑制侵袭、迁移和 EMT。
