Regulation of MIR100HG by SP1 transcription factor in hepatocellular cancer

MIR100HG is a long non-coding RNA (lncRNA) with significant effects in cancer biology, shown to be highly expressed in various cancer types. This thesis investigates the cellular effects of MIR100HG overexpression and suppression under normoxic and hypoxic conditions, the transcriptional activation of MIR100HG by SP1, and SP1 binding sites within the MIR100HG promoter using ChIP assays. The cellular and molecular effects of SP1 under normoxic and hypoxic conditions were also examined. In Hep3B cells, overexpression and stable suppression of MIR100HG were confirmed at the mRNA level. Under normoxic and hypoxic conditions, MIR100HG overexpression enhanced cell proliferation, migration, colony formation, and invasion potential. Conversely, lentiviral-based suppression of MIR100HG via shRNA reduced these capabilities under both conditions. SP1, a transcription factor with multiple binding sites in the MIR100HG promoter, was shown to regulate MIR100HG transcription. SP1 overexpression increased the activity of truncated promoter fragments, confirmed at the mRNA level via qRT-PCR. ChIP assays demonstrated SP1 binding to the -260/+56 promoter region. In Hep3B cells, SP1 overexpression and suppression were analyzed under normoxic and hypoxic conditions. Both were confirmed at mRNA and protein levels. SP1 overexpression decreased cell proliferation, migration, and colony formation, while stable suppression via shRNA increased these activities. SP1's effects on metastatic gene expression were assessed at the mRNA level.