Research Data presentaion during Maters/PhD defense.

Guide about making ppt for presenting research data during master or phd defense in life sciences.

1. Presentation for PhD Admission 25-03-2024

2. SELF INTRODUCTION Major; Chemical Engineering and Technology College; Life Science Supervisor; Changyuan Yu Co-supervisor; Yang Zhao MS Research Area ; Cancer Treatment ZUFA SABEEL (2018220039)

3. Screening Potential Drugs for Bladder Cancer Treatment and Uncovering Their Molecular Mechanisms of Action

4. Contents Introduction Background Objectives Methodology Result discussion of project 1 Result discussion of project 2 Limitation of current study Conclusion

5. Introduction Bladder cancer (BC) happens when cells in the bladder grow out of control and form tumors BC MIBC NMIBC Symptoms Causes and Risk Factors Survival Rates Diagnosis Treatment by Stage Prevention Blood in the urine Burning sensations Frequent & painful urination Lower back pain Pelvic pain Average overall – 77% In situ – 96% Localized – 69% Regional – 37% Distant – 6% Physical Exam Urinalysis Cystoscopy Cytology Intravenous Pyelogram Biopsy TURBT BCG immunothera-py Surgery Chemotherapy Radiation Immunothera-py & surgery Smoking Chronic UT infections Exposure to workplace chemicals Genetics RT to the pelvis Avoid smoking Limit chemical exposure Drink plenty of fluids

6. Background Curing bladder cancer is crucial for several reasons, and the need for a cure is underscored by the following background information Prevalence and Impact Limited Treatment Options Human Suffering Lack of Targeted Therapies Economic Burden Research and Innovation Unmet Medical Needs Impact on Quality of Life High Recurrence Rates

7. Drugs Chosen Paeonol (PAE) AR-A014418 a natural phenolic compound (Chinese peony tree) a synthetic small molecule (GSK-3 β inhibitor) potential biological and pharmacological properties Allows researchers to study the effects of GSK-3β inhibition on various cellular functions and signaling pathways inhibition of cell proliferation, suppression of invasion and migration, induction of apoptosis and cell cycle arrest Inhibition of GSK-3β resulted in a reduction of survival and proliferation in gastric cancer cells Inhibition of GSK-3β has been explored as a potential therapeutic strategy in various medical contexts, including cancer pancreas, colorectal, hepatocellular carcinoma, kidney, leukemia, and glioblastoma multiforme

8. Objectives To comprehensively investigate and understand the inhibitory effects and underlying mechanisms of two distinct compounds, Paeonol (PAE) and AR-A014418, on bladder cancer cells

9. Methodology

10. Result Discussion of Project 1

11. CCK-8 IC 50 （μM） T24 J82 24 h 472.9 μg/mL 453.9 μg/mL 48 h 212.4 μg/mL 123.7 μg/mL PAE Inhibits Bladder Cancer Cell Proliferation

12. Colony Formation Assay PAE Reduced Clonogenic Survival of Bladder Cancer Cells ****P<0.0001

13. PAE-Induced S-Phase Arrest in Cultured Bladder Cancer Cells ****P<0.0001 ****P<0.0001

14. Data is presented as the mean ± standard deviation from three independent experiments (n =3). Statistical significance: P < 0.05, ****P < 0.0001, PAE-treated vs. control (DMSO).

15. Induction of Apoptosis by PAE in Cultured Bladder Cancer Cells **p < 0.01 *p < 0.05

16. Effect of PAE on Transcriptional Regulation of Apoptosis-Related Genes in BC Cells Data are shown as mean ± SD of three independent experiments. Statistical significance: **P < 0.01, ***P<0.001, and ****P < 0.0001, PAE-treated vs. control (DMSO)

17. Effect of PAE on Translational Regulation of Apoptosis-Related Genes in BC Cells Data are shown as mean ± SD of three independent experiments. Statistical significance: **P < 0.01, ***P<0.001, and ****P < 0.0001, PAE-treated vs. control (DMSO) WB Study 1; Effect of PAE on BC Cells

18. AR-A014418 Suppresses Bladder Cancer Growth via NFκB and GSK-3β Regulation Result Discussion of Project 2

19. IC 50 （μM） EJ T24 BIU 87 24 h 70.06 µM 130.00 µM 356.45 µM 48 h 33.42 µM 53.13 µM 93.4 µM Cell Proliferation Assay to Determine the Optimal Inhibitory Concentration ****p<0.0001 compared with control was considered statistically significant

20. Drug combination and Dose-Effect Analysis of AR-A14418 and Cisplatin on Bladder Cancer Cells by CompuSyn Simulation Drug combination analysis of cisplatin and AR-A014418. (A) Dose-Effect Curve for combined drug treatment on bladder cancer. (B) Drug Reduction Index Plot for Combo: AR+Cis ( AR-A+Cis [10:1]). 1 in Y-axis denotes no drug reduction and all data point below 1 means this drug combination is not suitable and have no synergy on bladder cancer. (C) Logarithmic Combination Index Plot for AR-A014418 and cisplatin combo Total Dose Fa CI Value 55.0 0.127 2.63781 110.0 0.121 2.49285 220.0 0.126 9.15080 CI values for actual experimental points of AR-A014418 and cisplatin

21. Effect of AR-A014418 Inhibitor on the Cell Cycle of BC Cell Lines Effects of AR-A014418 on EJ cell cycle at 30 µM concentration. (A) Following a 48-hour incubation with AR-A,014418 flow cytometry was employed to assess cellular DNA content. (B) Chart depicting the distribution of cell cycle phases in EJ cells. Data is presented as the mean ± standard deviation from three independent experiments (n =3). ****P<0.0001

22. Effects of AR-A014418 inhibitor on Apoptosis of Bladder Cancer Cell Lines ***P=0.0004 ****P<0.0001

23. Induction of Cell Cycle Arrest and Apoptosis by Inhibited Expression of GSK-3β and NFκB in Bladder Cancer Treated by AR-A014418 Histograms show the change of relative protein expression of NFkB and GSK-3β in BC cells. Statistical significance: ***P<0.001, and ****P < 0.0001, AR-A014418-treated vs. control Study 2; Effect of AR-A014418 on BC Cells

24. AR-A014418 induced apoptotic cell death in Bladder Cancer Cells via upregulation of Bax, Bak, and Caspase-3 Histograms show the change of relative mRNA expression of apoptosis-related gene in BC cells after AR-A014418 treatment. Statistical significance: ****P <0.0001, treated vs. control (DMSO). Study 2; Effect of AR-A014418 on BC Cells

25. AR-A014418 induced apoptotic cell death in Bladder Cancer Cells via upregulation of Bax, Bak, and Caspase-3 Histograms show the change of relative protein expression of Bak, Bax and Casp-3 in BC cells. Statistical significance: **P < 0.01, ***P<0.001, and ****P < 0.0001, AR-A014418-treated vs. control Study 2; Effect of AR-A014418 on BC Cells

26. Study 2; Effect of AR-A014418 on BC Cells Effect of AR-A014418 (GSK-3β Inhibitor) on Cell Migration of Bladder Cancer

27. Limitation of Current Study Involving in vivo models or clinical trials, can build upon your in vitro work to provide a more comprehensive understanding of the drug's effectiveness and safety for bladder cancer treatment

28. In summary, the study found that Paeonol (PAE) exhibited significant inhibitory effects on bladder cancer cells, inducing cell cycle arrest and promoting apoptosis. This highlights PAE's potential as a promising therapeutic agent for bladder cancer. Additionally, AR-A014418, a small molecule inhibitor, demonstrated notable anti-proliferative and anti-migratory effects on bladder cancer cells. It induced cell cycle arrest and apoptosis by downregulating GSK-3beta and NFκB while upregulating pro-apoptotic genes and proteins. These findings suggest that AR-A014418 has multifaceted therapeutic potential in the treatment of bladder cancer. Conclusion

29. Publications As a first Author Sabeel Z ., Liang Y., Hao M., Ying L., Guo R., Chen R., et al. (2023). A comprehensive Review of antitumor properties of Angelica species and their antitumor-responsible constituents and the underlying molecular mechanisms involved in tumor inhibition . Phytotherapy Research. 37, 2187–2211. 10.1002/ptr.7841 As Co-Author 2. Ruolan Chen, Youfeng Liang, Lu Ying, Zufa Sabeel , Xiaoning Li, Mingxuan Hao, Rui Gou, Yongchao Li, Zhao Yang. “Research Progress of Human Origins”. Scientific and Social Research. https://doi.org/10.26689/ssr.v5i9