Pathological function of MUC16 in PC growth and metastasis

• MUC16 is the largest known mucin family glycoprotein that is expressed by the
various epithelial cells

• The transmembrane protein is of about 12000 amino acids consist of N-terminal

domain, tandem repeats and c-terminal domain.

• In addition to its normal protective function, deregulated expression of MUC16 has

been implicated in several pathological conditions such as cancer.

Pancreatic cancer

• The expression of MUC16 increases gradually with pancreatic cancer progression.

• Expression of MUC16 is significantly higher in moderate and poorly differentiated

when compared to well differentiated pancreatic cancer.

• Further it is shown that the expression of MUC16 is higher in metastatic tissues

when compared to the primary sites of same patients.

• All these suggest that MUC16 might play an important role in pancreatic cancer
progression and metastasis.
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 Interim summary

Collectively, our results show that MUC16 is involved in regulation of the

invasive phenotype of PC cells through promoting a new signalling pathway,
which is mediated by FAK/Akt/ERK, and the induction of EMT biomarkers
which enhance cancer cell motility and metastatic potential.

Thus, MUC16 could represent a novel potential therapeutic target in pancreatic cancer.

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 MUC16 interact with BM to promote metastasis
• In the processes of invasion and metastasis, tumor cells leave their primary site,
invade the surrounding extracellular matrix, penetrate the blood and lymph vessels,
and finally attach and proliferate at a secondary site.
• However, little is known of the mechanism of MUC16 that contribute to this cascade
of events that leads to local tumor invasion and the formation of distant
metastases.
• Previous studies from our lab has shown that MUC4, another Mucin interact with
galectin 3 and suggests that clustering of MUC4 on the cell surface may expose the
surface adhesion molecules, which in turn promotes a stronger attachment of
tumor cells to the endothelial surface.
• Further recent studies shown that
– MUC16 serves as a ligand for P and E selectins in pancreatic cancer cells and
aids in cancer cell metastasis
– MUC16 interacts with mesothelin in ovarian and pancreatic cancer cells to
facilitate metastasis.
– MUC16 interact with NK cells via the Siglec-9 receptor in patients with ovarian
cancer and induces immune suppression.
– Further MUC16 can interact with galectin 1 and 3 in normal epithelial cells to
contribute toward the formation of the dense glycocalyx, while in cancer cells,
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Galectins

•Galectins is a multifunctional b-galactoside binding lectin.

•Galections has been reported to be present in the nucleus, cytoplasm as well as

secreted outside the cell by a non-classical pathway and hence known to become a
part of the cell surface and the ECM [25,26].

•Galectin-3 and Galectin-1 has also been shown to be present in higher amounts in
pancreatic cancer cells and tissues.

•Intracellular galectin-3 has been implicated in several processes such as cell growth
and differentiation, regulation of apoptosis, whereas, extracellular galectin-3
promotes processes like cell–cell recognition, adhesion, invasion and metastasis
[26,27].

•While about galectin 1

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MUC16 interacts with Galectin-1 and Galectin-3 in colo 357 PC cells

Galectin-binding assay
***

*
Absorbance (485 /525 nm)

Galectin-binding assay
Briefly, MUC16 knockdown Colo-357 cells were plated in triplicate to galectin-1 and galectin-3 protein–coated 96-well plates. After 1hr
incubation at 37°C, unattached cells were washed carefully with PBS twice. The adhered cells were incubated with Calcein-AM dye for 1
hour at 37C. The relative fluorescence was measured at an excitation wavelength of 485 nm and the emission wavelength of 520 nm. P
values lower than 0.05 were considered statistically significant.

MUC16 interacts with both Galectin 1 and Galectin 3 in colo 357 pancreatic cancer cells.
In order to examine the possible interaction between MUC16 and galectins, MUC16 knockdown colo 357 cells were
incubated in a galectin coated plates for an hour and stained with calcein AM dye as described above. Fluorescent intensity analysis
revealed that MUC16 knockdown cells shows significant decrease in binding to both galectin-1 and galectin-3 recombinant protein.
However, binding to galectin-3 is more significant when compared to galectin -1 (Fig. ).
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 Future directions

• Establishment of MUC16 knockdown cells

• Determination of MUC16 interaction with specific BM proteins and will

determine is there any correlation with clinical progression

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