Reference no: EM132927540

1) What do the following abbreviations stand for?
i) O-GlcNAc
ii) PDAC
iii) HBP

2) Briefly describe the typical metabolic phenotype of cancer cells explained by the Warburg effect and how this may affect cellular levels of UDP-GlcNAc
3) Which amino acids can undergo post-translational modification by O-GlcNAcylation?

4) What previous work by the authors led them to consider the relationship between O-GlcNAcylation and pancreatic cancer cell survival?

5) Why did the authors investigate the relationship between pancreatic cancer cell proliferation and O-GlcNAcylation of NF-kB?

6) Produce a flow draw diagram to show how the transcription factor NF-kB is activated by IKK.

7) Explain the purpose of using the inhibitors Ac-5SGlcNAc and NButGT.

8) From where did the authors obtain the HPDE cells?

9) Explain how the FITC-Annexin V/propidium iodide apoptosis assay can distinguish between early/late apoptotic, necrotic and non-apoptotic cells.

10) What are shOGT1 and shOGT2 and how do they affect gene expression?

11) In your own words and with reference to the results in Figure 2, explain how the authors show that hyper-O-GlcNAcylation contributes to PDAC cell proliferation and anchorage-independent growth.

12) How do the authors show that hyper-O-GlcNAcylation contributes to tumour growth in vivo?

13) Figure 4B appears to show different effects of shOGT1 and shOGT2 on the induction of apoptosis in MiaPaCa-2 cells. How do the authors explain this and which other results presented in the paper do you think could support this view?

14) Why did the authors investigate the effect of hyper-O-GlcNAcylation on suspension-induced apoptosis?

15) In your own words explain what conclusions can be drawn from Figure 5 A-E

16) What was the purpose of detecting the following proteins in the Western blot result shown in Figure 5F:
i) Cyclin D1
ii) Bcl-xL
iii) β-actin

17) How do the authors determine that site specific O-GlcNAcylation of p65 contributes to NF-βB-dependent cell growth in soft agar cultures?

18) What evidence do the authors provide to suggest that the effects of hyper-O-GlcNAcylation on cell survival are specific to PDAC cells compared to HPDE cells and what mechanism do they propose may partly explain this observation?

19) Which previous study do the authors cite to support their comment that activation of IKKβ is enhanced by O¬-GlcNAc modification in p53-deficient cells?

20) How do you think the results from this study could translate into new clinical therapies and are there are any potential limitations to how successful such a treatment could be for pancreatic cancer?