Chen Lin, a teacher of Xiangya Second Hospital of Central South University, has long been engaged in the development and treatment of oral cancer. Recently, its laboratory discovered a circRNA molecule, circRNA_100290, that regulates the development of oral squamous cell carcinoma through the Arraystar circRNA chip. CircRNA_100290 binds miR-29 by endogenous competition, thereby inhibiting the inhibition of CDK6 by miR-29 and promoting the development of oral squamous cell carcinoma. The results were published in the Nature issue of Oncogene (IF = 7.932). (The chip is provided by Kang Cheng Biotech) .
Research Background
Oral squamous cell carcinoma (OSCC) is one of the top eight malignant tumors in the world, and the incidence and mortality are not reduced due to the improvement of surgical techniques. It is a recognized health problem. In the past decade, miRNA and lncRNA have been shown to play an important role in the development of cancer. In some articles, miRNAs can be used as regulators of OSCC. However, circRNAs are widely present in animals. The occurrence and progress of research are not very common, especially OSCC. At present, some circRNA studies such as ciRs-7, sry and circHIPK3 have been proved. By studying the circRNA and mRNA expression profiles of OSCC, the differentially expressed circRNA_100290 was obtained, and the corresponding target gene CDK6 was obtained by ceRNA mechanism analysis, and it was proved that it binds to miR-29 through endogenous competition, thereby regulating CDK6 expression and promoting OSCC. Development takes place to provide potential targets for the treatment of OSCC.
Research ideas
In order to explore the role of circRNA in the development and progression of OSCC, Arraystar circRNA microarray was used to analyze OSCC and NCMT samples, and the expression profile of circRNA was analyzed. It was found that a total of 280 circRNA expressions were significantly expressed, of which 139 circRNAs were up-regulated and 141 circRNAs were down-regulated. CircRNA_100290 was selected from the top 10 most important circRNAs, and q-PCR verified that circRNA_100290 did significantly up-regulate expression in OSCC tissues.
The circRNA has been reported to function through the ceRNA mechanism. The authors further explored the molecular mechanism of circRNA_100290, and predicted the circRNA_100290 target gene through the miRNA databases: Targetscan, miRanda and RNA22, and obtained the highest score of three microRNAs: miR-29b-3p. miR-29c-3p, miR-29a-3p and the most likely related target gene, CDK6, is a cell cycle-associated protein and is significantly up-regulated in expression profile chips. By knocking down the expression of the OSCC cell line circRNA_100290, the authors found that CDK6 expression was also significantly down-regulated in these cells, demonstrating that CDK6 expression is regulated by circRNA_100290. CDK6 plays a very important role in the cell cycle and promotes the entry of cells into the S phase of DNA synthesis. It has been demonstrated by MTT and BrdU experiments that knockout of circRNA_100290, or the use of miR-29 analogs, can significantly reduce the proliferation of CAL27 cells. The authors also verified in vivo levels that normal HN4 cells and circRNA_100290 knockout HN4 cells were implanted into mice. After 3 weeks, the unexpanded mice showed more tumor volume increase than circRNA_100290 knockdown. In addition to mice, luciferase assays demonstrated that miR-29 binds directly to circRNA_100290 and CDK6 in cells.
Technical route
Result display
Figure 1: CircRNA microarray screening for significant circRNA
Figure 2: ceRNA analysis reveals that circRNA_100290 regulates CDK6 via the mechanism of CeRNA
Figure 3: In vitro and in vivo validation of circRNA affects cell proliferation
Figure 4: Luciferase and EGFP/RFP reporter assays verify that miR-29 binds directly to circRNA_100290/CDK
Significance
The study revealed that circRNA_100290 binds to miR-29 by endogenous competition, regulates the expression of CDK6 gene, and thus affects the proliferation of oral squamous cell carcinoma, and can be used as a potential target for the treatment of OSCC.
about the author
Chen Lin, Ph.D., deputy chief physician of Oral and Maxillofacial Surgery, Second Xiangya Hospital, Central South University. He has long been engaged in clinical and research work in oral and maxillofacial surgery, and has extensive experience in the prevention and treatment of oral cancer. He has published dozens of articles related to oral clinical research.
Paper link
circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family.
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