TY - JOUR
T1 - Cancer-associated fibroblast (CAF) secretomes-induced epithelial-mesenchymal transition on HT-29 colorectal carcinoma cells associated with hepatocyte growth factor (HGF) signalling
AU - Wanandi, Septelia Inawati
AU - Hilbertina, Noza
AU - Siregar, Nurjati Chairani
AU - Abdullah, Murdani
AU - Jeo, Wifanto Saditya
N1 - Funding Information:
Funding Disclosure: This study was supported by a PDUPT grant from the Ministry of Research, Technology and Higher Education of the Republic of Indonesia.
Publisher Copyright:
© 2021 Pakistan Medical Association. All rights reserved.
PY - 2021/2/1
Y1 - 2021/2/1
N2 - OBJECTIVE: The aim of this study was to investigate the effect of cancer-associated fibroblasts (CAF) secretomes on the epithelial-mesenchymal transition (EMT) of colorectal carcinoma (CRC) cells and its association with hepatocyte growth factor (HGF) signalling focussing on the HGF receptor, c-Mesenchymal epithelial transition (c-Met), and the EMT markers, vimentin and e-cadherin, in CRC cells. Methods: Conditioned mediums (CM) containing secretomes from colorectal CAFs and their counterpart normal fibroblasts (NFs) of three CRC patients were collected and supplemented to the HT-29 CRC cells. The mRNA levels of a-smooth muscle actin (a-SMA) and HGF in both fibroblasts, as well as c-Met, vimentin, and e-cadherin in HT-29 cells after supplemented with CAF- and NF-CM were determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). HGF protein level in the CM of CAFs and NFs was measured using enzyme-linked immunosorbent assay (ELISA). Vimentin and e-cadherin protein expressions were observed in HT-29 cells using immunofluorescent (IF) staining. RESULTS: Compared to the non-cancerous colon, fibroblasts from cancerous area of CRC substantially expressed higher mRNA levels of a-SMA, a CAF marker. The HGF mRNA expressions in CAFs and NFs were in line with the HGF protein level in the secretomes of both cells. CAF-CM increased c-Met and vimentin mRNA levels in HT-29 cells. Surprisingly, e-cadherin mRNA level in HT-29 cells was increased following CAF-CM supplementation. We also demonstrated the co-localization of e-cadherin and vimentin in the HT-29 cell cytoplasm. CONCLUSIONS: CAF secretomes of CRC promote a hybrid type of EMT associated with HGF signalling.
AB - OBJECTIVE: The aim of this study was to investigate the effect of cancer-associated fibroblasts (CAF) secretomes on the epithelial-mesenchymal transition (EMT) of colorectal carcinoma (CRC) cells and its association with hepatocyte growth factor (HGF) signalling focussing on the HGF receptor, c-Mesenchymal epithelial transition (c-Met), and the EMT markers, vimentin and e-cadherin, in CRC cells. Methods: Conditioned mediums (CM) containing secretomes from colorectal CAFs and their counterpart normal fibroblasts (NFs) of three CRC patients were collected and supplemented to the HT-29 CRC cells. The mRNA levels of a-smooth muscle actin (a-SMA) and HGF in both fibroblasts, as well as c-Met, vimentin, and e-cadherin in HT-29 cells after supplemented with CAF- and NF-CM were determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). HGF protein level in the CM of CAFs and NFs was measured using enzyme-linked immunosorbent assay (ELISA). Vimentin and e-cadherin protein expressions were observed in HT-29 cells using immunofluorescent (IF) staining. RESULTS: Compared to the non-cancerous colon, fibroblasts from cancerous area of CRC substantially expressed higher mRNA levels of a-SMA, a CAF marker. The HGF mRNA expressions in CAFs and NFs were in line with the HGF protein level in the secretomes of both cells. CAF-CM increased c-Met and vimentin mRNA levels in HT-29 cells. Surprisingly, e-cadherin mRNA level in HT-29 cells was increased following CAF-CM supplementation. We also demonstrated the co-localization of e-cadherin and vimentin in the HT-29 cell cytoplasm. CONCLUSIONS: CAF secretomes of CRC promote a hybrid type of EMT associated with HGF signalling.
KW - Cancer-associated fibroblasts, Epithelial-mesenchymal transition, Hepatocyte growth factor, c-Met.
UR - http://www.scopus.com/inward/record.url?scp=85103683891&partnerID=8YFLogxK
M3 - Article
C2 - 33785936
AN - SCOPUS:85103683891
SN - 0030-9982
VL - 71 2)
SP - S18-S24
JO - JPMA. The Journal of the Pakistan Medical Association
JF - JPMA. The Journal of the Pakistan Medical Association
IS - 2
ER -
