Department of Oncology, Affiliated Xuzhou Central Hospital, Southeast University, Xuzhou, Jiangsu 221009, P.R. China.
Department of Oncology, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Int J Mol Med. 2018 Dec;42(6):3562-3570. doi: 10.3892/ijmm.2018.3871. Epub 2018 Sep 11.
Ultraviolet radiation resistance‑associated gene (UVRAG) regulates autophagy by promoting the formation and maturation of autophagosomes. The aim of the present study was to investigate the effects of UVRAG and UVRAG‑targeting miRNA on the regulation of autophagy and apoptosis in gastric cancer (GC). TargetScan was used to predict that miR‑183 targets the 3'‑untranslated region (UTR) of UVRAG, while the interaction between miR‑183 and the 3'‑UTR of UVRAG was assessed using a dual luciferase reporter assay. Autophagy was induced in the GC cell line MKN28 by serum starvation. miR‑NC mimics, miR‑183 mimics, miR‑NC inhibitors and miR‑183 inhibitors were transfected into MKN28 cells, followed by assessment of the UVRAG expression, cell viability and cell apoptosis by western blotting, reverse transcription‑quantitative polymerase chain reaction, Cell Counting Kit‑8 and flow cytometry, respectively. It was demonstrated that autophagy resulted in miR‑183 downregulation and overexpression of UVRAG mRNA and protein in MKN28 cells. Transfection with miR‑183 mimic or inhibitor affected the expression of miR‑183 and, consequently, UVRAG. miR‑183 overexpression reversed the starvation‑induced inhibition of cell proliferation, while inhibition of miR‑183 reduced the proliferation of GC cells. miR‑183 overexpression reversed starvation‑induced apoptosis, while miR‑183 inhibition promoted starvation‑induced apoptosis in GC cells. Finally, miR‑183 overexpression attenuated starvation‑induced autophagy (LC‑3), apoptosis (Bax/Bcl‑2) and UVRAG expression, while miR‑183 inhibition exerted the opposite effects. In conclusion, the results of the present study indicated that miR‑183 inhibits starvation‑induced autophagy and apoptosis by targeting UVRAG in human GC cells.
紫外线辐射抗性相关基因 (UVRAG) 通过促进自噬体的形成和成熟来调节自噬。本研究旨在探讨 UVRAG 和 UVRAG 靶向 miRNA 对胃癌 (GC) 中自噬和细胞凋亡的调节作用。TargetScan 用于预测 miR-183 靶向 UVRAG 的 3'非翻译区 (UTR)，而 miR-183 与 UVRAG 的 3'UTR 之间的相互作用则通过双荧光素酶报告基因检测进行评估。通过血清饥饿诱导 GC 细胞系 MKN28 中的自噬。将 miR-NC 模拟物、miR-183 模拟物、miR-NC 抑制剂和 miR-183 抑制剂转染至 MKN28 细胞中，然后通过 Western blot、逆转录-定量聚合酶链反应、细胞计数试剂盒-8 和流式细胞术分别评估 UVRAG 表达、细胞活力和细胞凋亡。结果表明，自噬导致 MKN28 细胞中 miR-183 下调和 UVRAG mRNA 和蛋白的过表达。转染 miR-183 模拟物或抑制剂会影响 miR-183 的表达，进而影响 UVRAG 的表达。miR-183 过表达逆转了饥饿诱导的细胞增殖抑制，而 miR-183 抑制则降低了 GC 细胞的增殖。miR-183 过表达逆转了饥饿诱导的细胞凋亡，而 miR-183 抑制则促进了 GC 细胞的饥饿诱导凋亡。最后，miR-183 过表达减弱了饥饿诱导的自噬 (LC-3)、凋亡 (Bax/Bcl-2) 和 UVRAG 表达，而 miR-183 抑制则产生了相反的效果。综上所述，本研究结果表明，miR-183 通过靶向人类 GC 细胞中的 UVRAG 抑制饥饿诱导的自噬和凋亡。
