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International Journal of Bioprinting Kidney hydrogel print for renal cancer model

Figure 6. Immunofluorescence staining and semi-quantification. (A) Immunofluorescence images for CD44, vimentin, TWIST, CDH2, and CDH1
proteins of ACHN cells. (B–F) Semi-quantification analysis of CD44, vimentin, TWIST, CDH2, and CDH1 proteins. (g) Responses of ACHN cells to
sunitinib in 2D, 10% GelMA, and 10% GelMA-2% Kidney.
the impact of RCC cellular metastasis at the molecular level. grades; for this reason, it may be a valuable therapeutic
These analyses involved immunofluorescence staining marker for targeted therapies of RCC. 45,46 The 10%
and RT-qPCR of key EMT-related markers, including GelMA-2% Kidney sample also showed higher MMP2 and
Slug, Snail, MMP2, MMP9, CD44, ZEB1, TWIST, CDH2, MMP9 gene expression compared with other groups, a fact
CDH1, and vimentin. EMT is one of the most crucial that is consistent with the finding that the overexpression
steps in the RCC metastatic process, where the renal of Snail family can promote matrix metallopeptidase
epithelial cells acquire the phenotypical characteristics of (MMP) expression in RCC. MMPs play important
43
mesenchymal cells and show enhanced cellular migration, roles in tumorigenesis and cancer cell progression since
invasion, plasticity, and chemoresistance. RCC originates they degrade the main components of the ECM and
40
from the epithelial cells of the kidney, and the EMT process facilitate RCC cellular metastasis. 47,48 The gene and protein
can promote the migration and metastasis of RCC cells. 41,42 expression of other EMT-related markers, including CD44,
Our findings showed that the expression of Snail and Slug vimentin, and ZEB1, was also upregulated in ACHN cells
gene was increased in ACHN cells assigned to the 10% exposed to the 10% GelMA-2% Kidney solution; each
GelMA-2% Kidney treatment compared to other groups. of these has also been significantly correlated with RCC
The Snail family transcription factors have been recognized metastasis, survival, and recurrence. 49-53 The cadherin
as important mediators of EMT since they regulate the switch refers to the upregulation of CDH2 expression
expression of several other EMT-related markers, including and the downregulation of CDH1 expression, which was
CDH1, MMPs, and vimentin. Higher Snail expression observed for ACHN cells cultured in 10% GelMA-2%
43
has been found to be correlated with worse disease-free Kidney via immunofluorescence staining and has been
and disease-specific survival of RCC patients and may found to associate with metastasis and poor prognosis. 54,55
43
also be an independent adverse prognostic biomarker Given that culturing in 10% GelMA-2% Kidney
for the recurrence and survival of patients with localized significantly promoted ACHN metastasis, it was then used
clear cell RCC after nephrectomy. We also found that in the experiments for the 3D bioprinting of nephron.
44
ACHN cells had higher TWIST expression when cultured
with the 10% GelMA-2% Kidney sample. TWIST is a key The RNAseq was performed for ACHN cells cultured
transcription factor that confers tumor cells with CSC-like with 10% GelMA-2% Kidney as well as a 10% GelMA
characteristics and is also associated with higher tumor control culture. These results showed that RUNX1-

Volume 10 Issue 2 (2024) 290 doi: 10.36922/ijb.1413