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Effects of Moringa Oleifera on Anti-Cancer Activity of Oncolytic Vesicular Stomatitis Virus (VSV)
in Cervical Cancer Cells. HONORS THESIS
BY STEPHANIE IBARRA
Outline
• Background Information
• Purpose
• Approach
• Data
• Conclusion
Background Information
• Cancer – treatment, but no cure
• Intracellular/epigenetic alterations• Type I IFN Response
• Cervical cancer
• Human Papillomavirus (HPV-16, -18)
• Hysterectomy, chemotherapy, and radiation.
Background Information
• Oncolytic Virotherapy (OV)
• Anticancer viruses• Oncolytic Viruses/Agents
• Natural tropism for tumor cells• Cancer epigenetic changes
Background Information
• Oncolytic Viruses
• Target and kill cancer cells
• No harm to normal cells
• Control molecular cell machinery
• Two major classes
• Natural tropism for cancer cells and nonpathogenic to humans.
• Genetically manipulated vectors
Background Information
• Vesicular Stomatitis Virus (VSV)
• Low pathogenicity to humans
• 11Kb with 5 coding genes
• Matrix (M) protein
• Defective Type I IFN response
• rwt vs. rM51R-M strains
Background Information
• Moringa Oleifera (Moringa pterygosperma)
• Antitumor activity • Pro-apoptotic
• Anti-proliferative
• Low toxicity/safe on humans
Purpose
• The goal of this project is to test the effects of methanolic (MeOH) moringa oleifera extract and vesicular stomatitis viruses (VSV), both wild type (rwt) and mutated (rM51R-M), on three different cervical cancer cell lines (SiHa, HeLa, and C4-II) upon incubation after treatment and infection.
Approach
• Cervical carcinoma cell lines HeLa, SiHa, and C4-II
• VSV-GFP rwt and rM51R-M
Flow Cytometry ELISA
Data – Flow Cytometry
0
10
20
30
40
50
60
70
80
% G
ATE
D
HeLa + MeOH
Figure 1. The percentage of positive gated cells (infected cells) was similar
between rwt and M51R-M, with a relative small increase of infection with
increasing MeOH moringa extract in M51R-M HeLa cells.
0
20
40
60
80
100
120
MEA
N F
LUO
RES
CEN
CE
HeLa + MeOH
Figure 2. Concentration of virion production increased with increasing amounts
of MeOH moringa extract. Increased amounts of expressed GFP were present in
HeLa cells infected with M51R-M.
0
10
20
30
40
50
60
70
80
90
100
% G
ATE
D
SiHa + MeOH
Figure 3. The percentage of positive gated SiHa cells infected with rwt was high
regardless the addition of MeOH moringa extract. The percentage of infection of
SiHa cells with rM51R-M did increase upon addition of MeOH moringa extract
in comparison to the untreated sample.
0
20
40
60
80
100
120
140
160
180
200
MEA
N F
LUO
RES
CEN
CE
SiHa + MeOH
Figure 4. Concentration of viral progeny production increased with increasing
amounts of MeOH moringa extract for both rwt and rM51R-M infected cells.
Data – Flow Cytometry
Data – Flow Cytometry
Figure 5. Percentage of infected cells upon infection by both VSV strains rwt and
rM51R-M. Both viral samples achieved a linear infection with of cells with
increasing concentrations of MeOH moringa extract treatment.
0
20
40
60
80
100
120
MEA
N A
BSO
RB
AN
CE
C4-II + MeOH
Figure 6. Mean production of viral progeny was observed to increase with
increasing concentrations of MeOH moringa extract in both rwt and rM51R
samples. The samples infected with rM51R-M achieved a higher progeny
production than the rwt samples.
0
10
20
30
40
50
60
70
% G
ATE
D
C4-II + MeOH
Data – ELISA
0
20
40
60
80
100
120
140
MEA
N C
ON
CEN
TRA
TIO
N (
PG
/ML)
Human IL-6 - 1:1 Dilution
Figure 8. The concentration of IL-production was notoriously higher for the samples
infected with rM51R and treated with MeOH moringa extract. The rwt samples had a
similar production of IL-6 to that of the mock and thus were not affected upon
treatment.
Conclusion
Analysis of the effects of Moringa oleiferamethanolic extract on anti-cancer activity upon infection with VSV oncolytic virus in cervical cancer cells showed that treatment does not hinder the degree of viral replication in cells and can potentially promote oncolytic virotherapy
Thank You!
