Abstract
CCT is a rate limiting enzyme required for the synthesis of phosphatidylcholine through the CDP‐choline pathway, which is critical for cell proliferation. Anti‐cancer drugs have therefore been designed based on their ability to inhibit this enzyme especially CCTα isoform. The purpose of this study was to explore non toxic molecular techniques of inhibiting CCTα expression or activity which results in the killing of lung cancer cells. Briefly, p53 positive, A549 and negative, H1299 lung epithelial cells were transfected with 0–800 nM of CCTα siRNA sequences (A, B, and C) or control siRNA for 24 hrs and cell viability and toxicity analyzed by Cell Titer Blue (CTB) as well as lactosedehydrogenase (LDH) assay. Western Blot analysis showed that all the CCTα siRNA sequences significantly inhibited CCTα protein expression while Light Microscopy depicted that CCTα siRNA transfected cells were completely shrunken and had nuclei filled with dark granular structures similar to apoptotic bodies. At high concentrations of siRNA the cells were completely degraded showing signs of cell lyses. CTB and LDH assays revealed that CCTα siRNA transfected cells were significantly less viable. These results demonstrate that CCTα siRNAs knocked down CCTα protein resulting in cell death, implying that the sequences could be useful as anticancer agents.
Published Version
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