Y-S Lv1, C Wang, L-X Li, S Han, Y Li. 1. Medical Center for Human Reproduction, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China. cyliyuan@126.com.
Abstract
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.
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.