ABSTRACT
To illustrate the potential function of lncRNA SBF2-AS1 in the progression of colorectal cancer (CRC) and the molecular mechanism. The relative level of SBF2-AS1 in CRC tissues and cell lines was determined by qRT-PCR. Its level in CRC patients with different tumor stages and tumor sizes was examined. After the knockdown of SBF2-AS1, proliferative, invasive abilities and apoptotic rate of CRC cells were evaluated. The correlation between SBF2-AS1 and PTEN was analyzed in CRC tissues. Furthermore, the subcellular distribution of SBF2-AS1 was assessed. Through RIP and ChIP assay, the interaction between SBF2-AS1 and PTEN was identified. Finally, the involvement of PTEN in SBF2-AS1-mediated CRC progression was analyzed. SBF2-AS1 was upregulated in CRC tissues and cell lines. Its level remained higher in CRC with worse tumor stage and larger tumor size. Knockdown of SBF2-AS1 attenuated proliferative, invasive abilities, but induced apoptotic rate of SW480 and DLD1 cells. A negative correlation was identified between expression levels of SBF2-AS1 and PTEN in CRC tissues. PTEN level was negatively regulated by SBF2-AS1. Subcellular distribution analysis indicated that SBF2-AS1 was mainly expressed in the nucleus. Furthermore, the RIP assay proved the binding of SBF2-AS1 to EZH2 and SUZ12. Knockdown of SBF2-AS1 attenuated the recruitment ability of EZH2 to PTEN. Notably, inhibited proliferation by transfection of sh-SBF2-AS1 1# was partially reversed after co-transfection of sh-PTEN. LncRNA SBF2-AS1 is upregulated in CRC. Knocking down of lncRNA SBF2-AS1 inhibits proliferation, and invasion and induces apoptosis of colorectal cancer by interacting with EZH2 to downregulate PTEN level.
Subject(s)
Colorectal Neoplasms , MicroRNAs , RNA, Long Noncoding , Humans , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , MicroRNAs/genetics , Cell Line, Tumor , Neoplasm Invasiveness/genetics , Apoptosis/genetics , Cell Proliferation/genetics , Cell Movement/genetics , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Gene Expression Regulation, Neoplastic , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolismABSTRACT
PURPOSE: ß-elemene has a wide range of anticancer effects and can be used in a variety of cancer types. This study mainly explored its mechanism of action on TNBC cells and provided theoretical basis for the treatment of TNBC. METHODS: Firstly, TNBC cells were treated with different concentrations of ß-elemene, and screened out an appropriate concentration for subsequent research. Then, through the bioinformatics website, predicted genes that have a binding relationship with ß-elemene. Then, the overexpression vector of the selected gene was transfected into the cell. The effects of ß-elemene and its target genes on the proliferation and apoptosis of TNBC cells were detected by CCK-8, Edu assay, and flow cytometry, and the senescence of cells was determined by SA-ß-gal experiment. Western blotting was used to detect the expression of apoptosis and aging-related proteins. RESULTS: ß-elemene inhibited TNBC cell viability and proliferation in a concentration-dependent manner, and induces apoptosis and senescence. Through the screening of candidate genes, IGF1 was finally determined to be an effective target gene of ß-elemene. The expression level of IGF1 was decreased in cells treated with ß-elemene. Overexpression of IGF1 significantly alleviated ability of ß-elemene to inhibit cell viability, proliferation, and induced cell apoptosis and senescence. In addition, ß-elemene inhibited the expression of IGF1R and Bcl-2, and promoted the expression of Cleaved Caspase-3 and senescence-related proteins (p27, p16, p53 and p21), and these effects were reversed by overexpression of IGF1. CONCLUSION: ß-elemene induced apoptosis and senescence of triple-negative breast cancer cells through IGF1/IGF1R pathway.
