During DNA replication, Okazaki fragments are formed at the growing replication fork to make sure both DNA strands grow in the 5’-to-3’ direction. However, when the replication fork reaches to the end of the linear chromosome, there is no enough space to produce the RNA primer starting the last Okazaki fragment at the very tip of a linear DNA molecule. Thus, eukaryotes use a RNA-protein complex, telomerase, to solve the problem.
Figure 1-Western blot results of proteins extracted from cell samples treated with 8 Nutraceutical combos and untreated one (Control).
Figure 2-Gel Electrophoresis Results of RT-PCR.
Figure 3-Alternatively spliced TERT transcripts observed in this study.
Figure 4-Diagram of TERT protein motifs being responsible for its reverse transcriptase activity.
Telomerase is a Ribonucleo-protein polymerase that plays essential role during DNA replication and maintains the chromosome length. Telomerase Reverse Transcriptase (TERT) is the protein component of telomerase, whose activity is usually turned down to avoid the uncontrolled cell proliferation. While the increased TERT expression is always related with the immortality of cancer cells, thus the TERT gene is a potential target for tumor treatment. Nearly all the eukaryotic cells apply alternative splicing to expand the coding capability of the genome and nutraceuticals are supposed to induce alternatively spliced RNAs. Thus, in this study, we treat Caco-2 cells with different nutraceutical combos and find some Caco-2 cells treated with nutraceuticals express more non-deletion transcript variants, hence more full-length TERT protein.
 Konnikova L, Simeone MC, Kruger MM. et al. Signal transducer and activator of transcription 3 (STAT3) regulates human telomerase reverse transcriptase (hTERT) expression in human cancer and primary cells. Cancer Res. 2005;65:6516–20.
 Akincilar, Semih Can, Bilal Unal, and Vinay Tergaonkar. “Reactivation of telomerase in cancer.” Cellular and Molecular Life Sciences (2016): 1-12.
I would like to thank my Teaching Assistants, Anthony Evans and Catharina Grubaugh for all of their patient guidance, advices and suggestions, as well as all the valuable work they put in to assure the experiment went as smoothly as possible. Finally, I would also like to thank Dr. Berish Rubin for his help and support to make this project possible.
|This document was last modified 05/16/2016.|
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