Curcumin

Curcumin effects on migration and autophagy of A549 lung cancer cells

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

EXPERIMENTAL APPROACH

CONTEXT

TIMING & BUDGET

A. Transfection with a miRNA and a GFP reporter B. Curcumin treatment with 4 different concentrations and piperine C. Wound Healing Assay (migration) D. LC3 immunodetection (autophagy)

• Cancer diseases
• Curcumin as an anticancer drug
• Inhibition of metastasis and PI3K/AKT/mTor pathway

• Two weeks to perform experiments

• 1184.08€ instead of 1319.23€

AIMS

EXPECTED OUTCOME

Objective: show that curcumin + miRNA206 can block migration and increase autophagy of cancer cells Specific aims:

• Effective concentration of curcumin
• Cell migration inhibition
• Autophagy induction

• Decrease in migration of cancer cells

• Increase of cancer cells autophagy

REFS.

COLLAB.

ABSTRACT

Sup'Biotech Interactive Scientific Poster ©

Curcumin effects on migration and autophagy of A549 lung cancer cells

CONTEXT

Cancer diseases

Intrusive current treatments

A natural alternative curcumin

unusual proliferation of abnormal cells

• Extracted from the spice, curcuma
• Plant name: Curcuma longa
• Anti-cancerous potential

17%

83%

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

CONTEXT

Hypothesis: curcucmin effects on migration and autophagy

Migration blocked?no metastasis

PI3K/AKT/mTOR inhibited?autophagic cell death through LC3

The role of PI3K/AKT/mTOR pathway in the modulation of autophagy and the clearance of protein aggregates in neurodegeneration 2014EnglishDavid Heras-Sandoval, Jazmin M. Pérez-Rojas, Jacqueline Hernández-Damián, José Pedraza-Chaverr

https://fr.dreamstime.com/images-libres-droits-m%C3%A9tastase-image23837249

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

CONTEXT

Methodology

Results

But no direct link curcumin-autophagy mentioned

miRNA206 vector transfection

Curcumin treatment

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

OBJECTIVE AND AIMS

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

EXPERIMENTAL APPROACH

• Transfection

Lipid transfection Transfectant : Turbofectin 8.0 Conditions of incubation :

• 37°C
• 5% CO2
• 24h

plasmid with miRNA

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

EXPERIMENTAL APPROACH

• Curcumin treatment

Legend: NT : Non-transfected T : Transfected NC : Non treated with curcumine Cx : treated with different concentration of curcumine

Plate plan for our experiments

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

EXPERIMENTAL APPROACH

Wound Healing Assay for migration

• Scratching on wells containing treated cells
• Different concentrations of Curcumin+Piperine
• Comparison between untreated and treated cells

https://www.iprasense.com/migration-assay/

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

EXPERIMENTAL APPROACH

Immunodetection for autophagy

• Target : LC3

• Primary antibody : Anti-LC3

• Marker : RFP (Red Fluorescent Protein)

• Tracking of LC3 location

The Use of Immunofluorescence Microscopy and Enzyme-Linked Immunosorbent Assayas Complementary Techniques for Protein Identification in Artists' Materials 2006EnglishArlen Heginbotham, Victoria Millay, Michael Quick

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

TIMING & BUDGET

Gantt chart

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

TIMING & BUDGET

Budget

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

EXPECTED OUTCOME

We except several results

Curcumin inhibits migration and invasion of non-small cell lung cancer cells through up-regulation of miR-206 and suppression of PI3K/AKT/mTOR sinaling pathway 2020Naizhi Wang, Tao Feng, Xiaona Liu, Qin Liu

• Stop or Decrease of migration : by comparing the scratch of the healing wound assay to a control one

After the scratch

Untreated cells

Treated cells

• Confirm the restart of autophagy : the fluorescence should be gathered around an autophagosome if autophagy is occurring

Autophagy

No autophagy

Propose an effective combination of Curcumin/Piperin with miRNA206 for those two issues

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

ABSTRACT

Cancer is the first cause of death across developed countries and lung cancer has the highest mortality rate. Curcumin has shown effects on some cancers, the goal of our project is then to understand its involvement on in migration and autophagy of A549 lung cancer cells. To do so, we will transfect the cells with a miRNA206 and realize perform curcumin treatment, wound healing assay and immunofluorescence. Transfection is the key step of our work, but the reagents needed are expensive; this grant would help us to carry out our project.

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

COLLABORATORS

A huge thank you to :

• Doctor Vanessa PERDIZ
• Sup'Biotech, Cell Tech laboratory (CEA)

BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

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BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia

Curcumin effects on migration and autophagy of A549 lung cancer cells

BIBLIOGRAPHY

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BERAMICE Agyei, CHAMPOMIER Céline, FAYE Awa Thioub, LEGRAVEREND Clélia