Ectopic M Tert Expression In Mouse Embryonic Stem Cells Does Not Affect Differentiation But Confers Resistance To Differentiation And Stress Indu

872 words - 4 pages

This study by the National Cancer Centre of the Republic of Singapore was intended to examine the possible work done by telomerase other than that of the extending of telomeres to maintain chromosomal integrity. A complex of the catalytic telomerase reverse transcriptase (TERT) and an RNA component forms the ribonucleoprotein that is telomerase which extends a sequence of TTTAGGG to the end of chromosomes in a reverse manner usual to biological dogma. Telomerase activity is down regulated as a cell specializes from stem cell to progenitor to terminal differentiation. This research was sparked by prior evidence that correlated telomerase with cellular differentiation and apoptosis and even ...view middle of the document...

Enzymatic activity for the corresponding lines was confirmed by western-blot analysis and TRAP assays. Overexpression of mTERT showed no difference in the rate of differentiation between the two cells lines over the course of losing all ‘stemness’ confirming TERT plays no part in regulating differentiation. With this observation in mind, the overexpressing colony of mTERT was noted to be significantly larger than the other cell line. Once again, markers were used to ensure differentiation rates were similar while apoptosis was measured via other markers. There was a significant difference in cell death in both undifferentiated and differentiated cells in comparison to the overexpressed and control lines. Furthermore, cell death was measured in cases of gentoxic insults such as chemotherapeutic and oxidative agents and overexpressing mTERT cells resulted in less cell death. In addition, catalytically inactive mTERT cells showed no difference in apoptosis compared to control cells using the same technique as before. Together, these experiments support resistance to apoptosis due to mTERT in both undifferentiated and differentiated cells in a variety of conditions.
Similar to the processes in growing overexpressed mTERT cells, cell lines were produced to have inactive p53 where p53 is a protein that can program cell death. Cells that did express inactive p53 showed similar anti-apoptosis results as overexpressed mTERT cells with active p53. mTERT was not shown to change p53 levels but instead affected downstream products of p53 showing similar results as alpha-PFT, an inhibitor that acts downstream of p53, in the absence of PFT-alpha. When both PFT and overexpressed mTERT were introduced simultaneously, an insignificant change in apoptosis occurred indicating similar functions. Together this data shows that mTERT protects against p53-dependent cell...

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