Q Wang1, P-M Kang. 1. Department of Respiratory and Critical Care Medicine, Shanxi Provincial People's Hospital, Taiyuan, China. qiangwang97@aliyun.com.
Abstract
OBJECTIVE: It has been demonstrated that circular RNA (circRNA) plays an important regulatory role in a series of diseases. The purpose of this study is to investigate the expression of circRNA_001010 and its facilitating effects on proliferation and invasion of non-small cell lung cancer (NSCLC) by regulating oncogene CDK4 through sponging with miR-5112. PATIENTS AND METHODS: qRT-PCR was performed to detect the expressions of circRNA_001010 and CDK4 in human NSCLC tissues and cells. Cell Counting Kit-8 (CCK-8) assay was performed to evaluate the A549 cells proliferation and transwell assay was performed to evaluate the A549 cells migration. Correlation analysis between circRNA_001010 and miR-5112 was detected by statistical analysis. Bioinformatics prediction was made to detect the binding site of GTL and miR-5112 and Luciferase activity was conducted to investigate the interaction between circRNA_001010 and miR-5112. Furthermore, we cloned the mice CDK4 3'-UTR into the Luciferase reporter vector and constructed miR-5112 binding mutants to validate the inhibited modulation of miR-5112 to the CDK4 expression. RESULTS: Results showed that the expressions of circRNA_001010 and CDK4 were upregulated in human NSCLC tissues and cells. qRT-PCR and CCK-8 assay showed that circRNA_001010 expression is associated with the proliferation of NSCLC cells, and that upregulated circRNA_001010 contributed to cell proliferation of A549. Transwell assay showed that circRNA_001010 was associated with the migration ability of tumor cells, and that increased expression of circRNA_001010 promoted the migration and invasion of NSCLC cells. The bioinformatics prediction and Luciferase assay demonstrated that by sponging with miR-5112, circRNA_001010 can serve as a ceRNA for miR-5112 to further regulate the expression of CDK4. CONCLUSIONS: For the first time, we found that circRNA_001010 was upregulated in human NSCLC patients, which could accelerate tumor proliferation, migration and invasion as a molecular sponge by modulating the inhibitory effect of miR-5112 on oncogene CDK4.
OBJECTIVE: It has been demonstrated that circular RNA (circRNA) plays an important regulatory role in a series of diseases. The purpose of this study is to investigate the expression of circRNA_001010 and its facilitating effects on proliferation and invasion of non-small cell lung cancer (NSCLC) by regulating oncogene CDK4 through sponging with miR-5112. PATIENTS AND METHODS: qRT-PCR was performed to detect the expressions of circRNA_001010 and CDK4 in humanNSCLC tissues and cells. Cell Counting Kit-8 (CCK-8) assay was performed to evaluate the A549 cells proliferation and transwell assay was performed to evaluate the A549 cells migration. Correlation analysis between circRNA_001010 and miR-5112 was detected by statistical analysis. Bioinformatics prediction was made to detect the binding site of GTL and miR-5112 and Luciferase activity was conducted to investigate the interaction between circRNA_001010 and miR-5112. Furthermore, we cloned the miceCDK4 3'-UTR into the Luciferase reporter vector and constructed miR-5112 binding mutants to validate the inhibited modulation of miR-5112 to the CDK4 expression. RESULTS: Results showed that the expressions of circRNA_001010 and CDK4 were upregulated in humanNSCLC tissues and cells. qRT-PCR and CCK-8 assay showed that circRNA_001010 expression is associated with the proliferation of NSCLC cells, and that upregulated circRNA_001010 contributed to cell proliferation of A549. Transwell assay showed that circRNA_001010 was associated with the migration ability of tumor cells, and that increased expression of circRNA_001010 promoted the migration and invasion of NSCLC cells. The bioinformatics prediction and Luciferase assay demonstrated that by sponging with miR-5112, circRNA_001010 can serve as a ceRNA for miR-5112 to further regulate the expression of CDK4. CONCLUSIONS: For the first time, we found that circRNA_001010 was upregulated in humanNSCLCpatients, which could accelerate tumor proliferation, migration and invasion as a molecular sponge by modulating the inhibitory effect of miR-5112 on oncogene CDK4.