环状 RNA_103993 通过 miR-1271/ERG 信号通路对非小细胞肺癌细胞增殖和凋亡的影响。
Effects of circRNA_103993 on the proliferation and apoptosis of NSCLC cells through miR-1271/ERG signaling pathway.
机构信息
Medical Center for Human Reproduction, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Aug;24(16):8384-8393. doi: 10.26355/eurrev_202008_22635.
OBJECTIVE
Non-small cell lung cancer (NSCLC) accounts for more than 80% of lung cancer. CircRNA is a new type of non-coding RNA. CircRNA was found to be deeply involved in the regulation of NSCLC cells. However, the principle of CircRNA regulating NSCLC cells needs to be further explored.
MATERIALS AND METHODS
The relative mRNA expression levels of CircRNA_103993, miR-1271 and ERG were detected by quantificational real-time polymerase chain reaction (qRT-PCR) in NSCLC cells and human bronchial epithelial cell line HBE. Cell proliferation was detected by the Cell Counting Kit (CCK8) when NSCLC cells were transfected with si-CircRNA_103993, si-NC, miR-1271 mimics, miR-NC, LV-ERG, respectively. The apoptotic rate of NSCLC cells was measured by apoptotic assay and flow Cytometry. The relative mRNA and protein expression levels of PCNA and caspase-3 were detected by Western blot and qRT-PCR.
RESULTS
CircRNA_103993 and ERG were significantly up-regulated while miR-1271 was significantly down-regulated in NSCLC cells. Knockdown of CircRNA_103993 and high expression of miR-1271 significantly inhibited NSCLC cell proliferation and promoted apoptosis. The double luciferin report showed that CircRNA_103993 served as a sponge of miR-1271 and miR-1271 could directly target ERG in NSCLC cells. More importantly, low expression of CircRNA_103993 increased the expression level of miR-1271, and high expression of miR-1271 decreased the expression level of ERG. Further experiments showed that miR-1271 inhibitors reversed the effect of si-CircRNA_103993 on proliferation and apoptosis of NSCLC cells. However, miR-1271 mimics significantly promoted the effects of si-CircRNA_103993 on the proliferation and apoptosis. Moreover, LV-ERG reversed the effect of miR-1271 mimics on proliferation and apoptosis of NSCLC cells. However, si-ERG significantly promoted the effects of miR-1271 mimics on the proliferation and apoptosis.
CONCLUSIONS
CircRNA_103993 was highly expressed in NSCLC cells. CircRNA_103993 regulated proliferation and apoptosis of NSCLC cells by acting as a sponge of miR-1271. The CircRNA_103993 /miR-1271/ ERG axis had an important effect on the proliferation and apoptosis of NSCLC cells. Therefore, CircRNA_103993 may be a target for treating lung cancer.
目的
非小细胞肺癌（NSCLC）占肺癌的 80%以上。CircRNA 是一种新型的非编码 RNA。已经发现 CircRNA 深度参与了 NSCLC 细胞的调控。然而，CircRNA 调控 NSCLC 细胞的原理需要进一步探索。
材料和方法
通过实时定量聚合酶链反应（qRT-PCR）检测 NSCLC 细胞和人支气管上皮细胞系 HBE 中 CircRNA_103993、miR-1271 和 ERG 的相对 mRNA 表达水平。当 NSCLC 细胞转染 si-CircRNA_103993、si-NC、miR-1271 模拟物、miR-NC、LV-ERG 时，通过细胞计数试剂盒（CCK8）检测细胞增殖。通过凋亡检测和流式细胞术测量 NSCLC 细胞的凋亡率。通过 Western blot 和 qRT-PCR 检测 PCNA 和 caspase-3 的相对 mRNA 和蛋白表达水平。
结果
CircRNA_103993 和 ERG 在 NSCLC 细胞中显著上调，而 miR-1271 显著下调。CircRNA_103993 的敲低和 miR-1271 的高表达显著抑制 NSCLC 细胞增殖并促进凋亡。双荧光素酶报告表明 CircRNA_103993 作为 miR-1271 的海绵，miR-1271 可以直接靶向 NSCLC 细胞中的 ERG。更重要的是，CircRNA_103993 的低表达增加了 miR-1271 的表达水平，而 miR-1271 的高表达降低了 ERG 的表达水平。进一步的实验表明，miR-1271 抑制剂逆转了 si-CircRNA_103993 对 NSCLC 细胞增殖和凋亡的影响。然而，miR-1271 模拟物显著促进了 si-CircRNA_103993 对 NSCLC 细胞增殖和凋亡的作用。此外，LV-ERG 逆转了 miR-1271 模拟物对 NSCLC 细胞增殖和凋亡的作用。然而，si-ERG 显著促进了 miR-1271 模拟物对 NSCLC 细胞增殖和凋亡的作用。
结论
CircRNA_103993 在 NSCLC 细胞中高表达。CircRNA_103993 通过作为 miR-1271 的海绵调节 NSCLC 细胞的增殖和凋亡。CircRNA_103993/miR-1271/ERG 轴对 NSCLC 细胞的增殖和凋亡有重要影响。因此，CircRNA_103993 可能是治疗肺癌的一个靶点。
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