25 January 2019 : Laboratory Research
Berberine Promotes Apoptosis of Colorectal Cancer via Regulation of the Long Non-Coding RNA (lncRNA) Cancer Susceptibility Candidate 2 (CASC2)/AU-Binding Factor 1 (AUF1)/B-Cell CLL/Lymphoma 2 (Bcl-2) AxisWei Dai12ABCD, Liyuan Mu12ABCD*, Yali Cui12BCE, Yingying Li12BCD, Ping Chen12BCEF, Hongjian Xie12BC, Xia Wang12BCDE
Med Sci Monit 2019; 25:730-738
BACKGROUND: Berberine, a natural isoquinoline alkaloid derived from Berberis genus plants, has been reported to have anti-cancer effects. While cell behavior can be modulated by long non-coding RNAs (lncRNAs), the contributions of lncRNAs in progression and berberine effects on colorectal cancer are largely unknown. Therefore, the present study investigated the involvement and regulatory function of lncRNA cancer susceptibility candidate 2 (CASC2) during the treatment of human colorectal cancer using berberine.
MATERIAL AND METHODS: Reverse transcription-quantitative PCR (RT-qPCR) was performed to detect the expression levels of lncRNA CASC2 and Bcl-2 mRNA in colorectal cancer cells. MTT assay was performed to evaluate cell viability. Flow cytometry and TUNEL assay were used to analyze the apoptosis of cancer cells. RNA immunoprecipitation (RIP) assay was done to verify the interaction between lncRNA CASC2 and (AU-binding factor 1) AUF1, or AUF1 and B-cell CLL/lymphoma 2 (Bcl-2).
RESULTS: Treatment with berberine suppressed cell viability of colorectal cancer by promoting apoptosis level. LncRNA CASC2 was upregulated in cells treated with berberine, and knockdown of lncRNA CASC2 reversed the berberine-induced apoptosis. In addition, anti-apoptotic gene Bcl-2 was suppressed by berberine treatment and lncRNA CASC2, inducing the pro-apoptotic effects. Moreover, lncRNA CASC2 binds to AUF1, which sequestered AUF1 from binding to Bcl-2 mRNA, thus inducing the inactivation of Bcl-2 translation.
CONCLUSIONS: Our study reveals that lncRNA CASC2 mediates the berberine-induced pro-apoptotic effect via inhibition of Bcl-2 expression at the post-transcriptional level.
Keywords: Apoptosis, Barbering, Colorectal Neoplasms, Hereditary Nonpolyposis, RNA, Long Noncoding, Berberine, Cell Movement, Cell Proliferation, Colorectal Neoplasms, Disease Progression, Down-Regulation, Gene Expression Regulation, Neoplastic, HCT116 Cells, HT29 Cells, Heterogeneous Nuclear Ribonucleoprotein D0, Heterogeneous-Nuclear Ribonucleoprotein D, MicroRNAs, Proto-Oncogene Proteins c-bcl-2, RNA, Messenger, Tumor Suppressor Proteins, Up-Regulation
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