Abstract
We here tested expression and potential functions of circular RNA PRKCI (circPRKCI) in human glioma. Our results show that circPRKCI is upregulated in human glioma tissues and glioma cells, correlating with downregulation of its potential target, microRNA-545 (miR-545). In A172 and primary human glioma cells, shRNA-mediated silencing of circPRKCI inhibited cancer cell growth, survival, proliferation, and migration. Conversely, ectopic circPRKCI overexpression promoted A172 cell progression. miR-545 is the primary target of circPRKCI in glioma cells. Forced overexpression of miR-545 mimicked circPRKCI shRNA-induced actions, inhibiting glioma cell survival and proliferation. In contrast, miR-545 inhibition, by a lentiviral antagomiR-545 construct, reversed circPRKCI shRNA-induced anti-A172 cell activity. Importantly, neither circPRKCI shRNA nor circPRKCI overexpression was effective in miR-545-knockout (Cas9 method) A172 cells. Importantly, the subcutaneous and orthotopic A172 xenograft growth was significantly inhibited by circPRKCI silencing. Collectively, circPRKCI promotes human glioma cell progression possibly by inhibiting miR-545. Targeting circPRKCI-miR-545 cascade could efficiently inhibit human glioma cells.
Highlights
Glioma, the most common brain tumor and a global health threat, causes significant mortalities each year[1,2,3]
CircPRKCI is upregulated in A172 glioma cells and in the primary human glioma cells (“Pri-1/-2/-3”, see Methods) (Fig. 1b)
While its levels are low in primary human neuronal cultures and human astrocytes (Dr Cao19) (Fig. 1b)
Summary
The most common brain tumor and a global health threat, causes significant mortalities each year[1,2,3]. MiR-545 levels are significantly downregulated in glioma tissues (Fig. 1c), as well as in the established and primary human glioma cells (Fig. 1d). CircPRKCI shRNA inhibits A172 glioma cell growth, survival, proliferation, and migration GV248 lentiviral shRNA, targeting non-overlapping sequence (“Seq1/Seq-2”) against circPRKCI (“sh-circPRKCI”), was added to A172 glioma cells. These results show that circPRKCI silencing inhibited cell growth, survival, proliferation, and migration, whiling inducing apoptosis activation in A172 glioma cells.
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