Department of Oncology, Yancheng City No. 1 People's Hospital, Yancheng, China.
Eur Rev Med Pharmacol Sci. 2019 Jan;23(2):649-659. doi: 10.26355/eurrev_201901_16879.
Accumulating evidence has suggested that aberrant expression of microRNAs (miRNAs) is associated with non-small cell lung cancer (NSCLC) proliferation, migration, invasion and chemotherapy resistance. Cullin4A (CUL4A) has been previously reported to desensitize NSCLC cells to chemotherapy treatment. However, whether miRNAs regulate CUL4A to promote chemotherapy resistance remains unknown.
Tissues were obtained from 40 NSCLC patients who received surgery at the Yancheng City No. 1 People's Hospital. Cell Counting Kit-8 (CCK-8) assays were applied for the detection of cell proliferation; mRNA and protein levels were determined by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) and Western blot, respectively. The interaction between mRNA 3'UTR and miRNA was predicted by TargetScan and verified by Dual-Luciferase reporter assay.
In the present study, miR-363-3p levels were revealed to be significantly decreased in tumor tissues obtained from NSCLC patients compared with adjacent normal tissues. The results of the CCK-8 assays showed that the overexpression of miR-363-3p may slightly inhibit the proliferation of A549 and H23 cells. Notably, the transfection with miR-363-3p antagonists reduced the sensitivity of A549 and H23 cells to gemcitabine treatment, whereas the overexpression of miR-363-3p markedly increased the sensitivity of A549 and H23 cells to gemcitabine treatment. Furthermore, CUL4A mRNA and protein levels were revealed to be decreased in A549 cells transfected with miR-363-3p mimics. The Dual-Luciferase reporter assay results further suggested that CUL4A represents a target gene of miR-363-3p.
The results indicated that decreased miR-363-3p expression enhanced gemcitabine resistance in NSCLC cells via regulation of CUL4A.
越来越多的证据表明，微小 RNA（miRNA）的异常表达与非小细胞肺癌（NSCLC）的增殖、迁移、侵袭和化疗耐药有关。Cullin4A（CUL4A）先前已被报道使 NSCLC 细胞对化疗治疗产生耐药性。然而，miRNA 是否通过调节 CUL4A 来促进化疗耐药性尚不清楚。
从在盐城市第一人民医院接受手术的 40 名 NSCLC 患者中获取组织。应用细胞计数试剂盒-8（CCK-8）检测细胞增殖；通过实时定量聚合酶链反应（RT-qPCR）和 Western blot 分别测定 mRNA 和蛋白水平。通过 TargetScan 预测 mRNA 3'UTR 和 miRNA 之间的相互作用，并通过双荧光素酶报告基因检测进行验证。
在本研究中，与相邻正常组织相比，NSCLC 患者肿瘤组织中 miR-363-3p 的水平明显降低。CCK-8 检测结果表明，miR-363-3p 的过表达可能轻微抑制 A549 和 H23 细胞的增殖。值得注意的是，miR-363-3p 拮抗剂的转染降低了 A549 和 H23 细胞对吉西他滨治疗的敏感性，而 miR-363-3p 的过表达显著增加了 A549 和 H23 细胞对吉西他滨治疗的敏感性。此外，转染 miR-363-3p 模拟物的 A549 细胞中 CUL4A mRNA 和蛋白水平降低。双荧光素酶报告基因检测结果进一步表明，CUL4A 是 miR-363-3p 的靶基因。
研究结果表明，miR-363-3p 表达下调通过调节 CUL4A 增强 NSCLC 细胞对吉西他滨的耐药性。
