BMI-1 Inhibition and Hippo Signaling in Alveolar Rhabdomyosarcoma Pubblico

Abstract

Alveolar rhabdomyosarcoma (ARMS) is a prevalent pediatric soft-tissue sarcoma with poorlow prognosis in high-risk patients. Recent research shows that BMI-1, an epigenetic histone repressor and stem cell factor, expresses a significant oncogenic effect within many cancers. Interestingly, one of these effects is on the cellular contact inhibition Hippo protein pathway. Utilizing western blot analysis, Annexin V/PI staining, and BrdU/7-AAD staining on siRNA-induced and PTC-028 drug-induced BMI-1 downregulated ARMS cells, we examined the effects of BMI-1 inhibition on Hippo pathway proteins and on cellular apoptosis and the cell cycle. We found that BMI-1 inhibition rescues the Hippo pathway at LATSats1/2 in the ARMS Rh30 cell line, upregulating phosphorylation at core Hippo pathway proteins LLATS1/2 ats1/2 and possibly YAP. Furthermore, in Rh28, PTC-028 mediated BMI-1 inhibition may turn on the Hippo pathway at MST1. BMI-1 inhibition by drug and siRNA has also shown an increase in apoptosis and decrease in proliferation in tumorigenic ARMS cells. Hence, study of cancer pathway effects of BMI-1 has validated its potential as a powerful target of PTC-028 drug therapy, providing a clinical method to restore  contact inhibition , cell cycle arrest and repair and apoptotic processes in pediatric rhabdomyosarcomas.

Table of Contents

Table of Contents

I.                   Introduction and Literature Review………………………………………………………….....1

A.     Introduction to Cancer…………………………………………………....……………1

B.     Overview of Rhabdomyosarcoma…………………………………....…….......3

C.     Novel epigenetic target, BMI-1……………………………………………....……6

D.     Overview of the Hippo Pathway…………………………………....…………….8

E.      BMI-1 Inhibitor, PTC-028……………………………………………....……….....10

F.      Hypotheses and Aims………………………………………………………....……..11

II.                 Materials and Methods……………………………………………………………………….......12

III.               Results........................................................................................................14

A.     Knockdown Determination of BMI-1..............................................14

B.     Determining BMI-1 interactions with the Hippo Pathway.......;.....16

C.     Determining effects of BMI-1 on overall cell cycle and vitality......20

IV.               Discussion.................................................................................................24

A.     siRNA Transfection Validation........................................................24

B.     BMI-1 Inhibition Turns oOn the Hippo Pathway................................25

C.     BMI-1 inhibition increases cellular apoptosis and diminishes progression to S-phase..............................................................................................28

V.                 Further Research and Conclusions.............................................................31

VI.               Bibliography..............................................................................................33

VII.             Non-Print References................................................................................36

I.                       Appendix A: Supplementary Figures..........................................................37

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About this Honors Thesis

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School	Emory College
Department	Biology
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Cell Health Sciences, Oncology Biology, Molecular
Parola chiave	BMI-1 Rhabdomyosarcoma Hippo
Committee Chair / Thesis Advisor	Robert Schnepp, Emory University
Committee Members	Jennifer Heemstra, Emory University Anita Corbett, Emory University

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