TY - JOUR
T1 - A mechanically-induced colon cancer cell population shows increased metastatic potential
AU - Tang, Xin
AU - Kuhlenschmidt, Theresa B.
AU - Li, Qian
AU - Ali, Shahjahan
AU - Lezmi, Stephane
AU - Chen, Hong
AU - Pires-Alves, Melissa
AU - Laegreid, William W.
AU - Saif, Taher A.
AU - Kuhlenschmidt, Mark S.
N1 - Funding Information:
X. T. was funded from NSF Grant 0965918 IGERT: Training the Next Generation of Researchers in Cellular and Molecular Mechanics and BioNanotechnology. We gratefully thank Mrs. Ketaki Bhide, Mr. Ravikiran Donthu, Dr. Jyothi Thimmapuram and Dr. Jenny Drnevich Zadeh for their effective help on RNA-Seq data processing at the University of Illinois Roy J. Carver Biotechnology Center. Insightful discussions with Prof. Norm E. Sladek (Masonic Cancer Center, Univ. of Minnesota), Prof. John Katzenellenbogen, and Prof. Fei Wang (UIUC) are highly appreciated. We also thank Dr. Mayandi Sivaguru for the invaluable training with immunocytochemical staining and confocal microscopy imaging at the Institute for Genomic Biology (IGB), UIUC. Assistance in PA gels preparations by Ms. Jana DiDomenico, Ms. Linna Guan, Ms. Vivian E Jung, Mr. Joe Laskowski and Mr. Stephan Lane are gratefully acknowledged. The work was supported by the grants: National Science Foundation 10– 02165, 0965918 IGERT, National Institute of Health RO1 083272–03, and UIUC Interdisciplinary Innovation Initiative Program grant #12035. We acknowledge the assistance of Dr. Scott MacLaren (Frederick Seitz Materials Research Laboratory (MRL), UIUC) with the atomic force microscopy (AFM) experiments, and Ms. Linna Guan (MSE, UIUC) and Ms. Vivian Jung (MCB, UIUC) with the preparation of PA gel substrates and cell culture. X. T. was funded from NSF Grant 0965918 IGERT: Training the Next Generation of Researchers in Cellular and Molecular Mechanics and BioNanotechnology. We gratefully thank Mrs. Ketaki Bhide, Mr. Ravikiran Donthu, Dr. Jyothi Thimmapuram and Dr. Jenny Drnevich Zadeh for their effective help on RNA-Seq data processing at the University of Illinois Roy J. Carver Biotechnology Center. Insightful discussions with Prof. Norm E. Sladek (Masonic Cancer Center, Univ. of Minnesota), Prof. John Katzenellenbogen, and Prof. Fei Wang (UIUC) are highly appreciated. We also thank Dr. Mayandi Sivaguru for the invaluable training with immunocytochemical staining and confocal microscopy imaging at the Institute for Genomic Biology (IGB), UIUC. Assistance in PA gels preparations by Ms. Jana DiDomenico, Ms. Linna Guan, Ms. Vivian E Jung, Mr. Joe Laskowski and Mr. Stephan Lane are gratefully acknowledged.
PY - 2014/5/29
Y1 - 2014/5/29
N2 - Background: Metastasis accounts for the majority of deaths from cancer. Although tumor microenvironment has been shown to have a significant impact on the initiation and/or promotion of metastasis, the mechanism remains elusive. We previously reported that HCT-8 colon cancer cells underwent a phenotypic transition from an adhesive epithelial type (E-cell) to a rounded dissociated type (R-cell) via soft substrate culture, which resembled the initiation of metastasis. The objective of current study was to investigate the molecular and metabolic mechanisms of the E-R transition.Methods: Global gene expressions of HCT-8 E and R cells were measured by RNA Sequencing (RNA-seq); and the results were further confirmed by real-time PCR. Reactive oxygen species (ROS), anoikis resistance, enzyme activity of aldehyde dehydrogenase 3 family, member A1 (ALDH3A1), and in vitro invasion assay were tested on both E and R cells. The deformability of HCT-8 E and R cells was measured by atomic force microscopy (AFM). To study the in vivo invasiveness of two cell types, athymic nude mice were intra-splenically injected with HCT-8 E or R cells and sacrificed after 9 weeks. Incidences of tumor development and metastasis were histologically evaluated and analyzed with Fisher's exact test.Results: Besides HCT-8, E-R transition on soft substrates was also seen in three other cancer cell lines (HCT116, SW480 colon and DU145 prostate cancer). The expression of some genes, such as ALDH3A1, TNS4, CLDN2, and AKR1B10, which are known to play important roles in cancer cell migration, invasion, proliferation and apoptosis, were increased in HCT-8 R cells. R cells also showed higher ALDH3A1 enzyme activity, higher ROS, higher anoikis resistance, and higher softness than E cells. More importantly, in vitro assay and in vivo animal models revealed that HCT-8 R cells were more invasive than E cells.Conclusions: Our comprehensive comparison of HCT-8 E and R cells revealed differences of molecular, phenotypical, and mechanical signatures between the two cell types. To our knowledge, this is the first study that explores the molecular mechanism of E-R transition, which may greatly increase our understanding of the mechanisms of cancer mechanical microenvironment and initiation of cancer metastasis.
AB - Background: Metastasis accounts for the majority of deaths from cancer. Although tumor microenvironment has been shown to have a significant impact on the initiation and/or promotion of metastasis, the mechanism remains elusive. We previously reported that HCT-8 colon cancer cells underwent a phenotypic transition from an adhesive epithelial type (E-cell) to a rounded dissociated type (R-cell) via soft substrate culture, which resembled the initiation of metastasis. The objective of current study was to investigate the molecular and metabolic mechanisms of the E-R transition.Methods: Global gene expressions of HCT-8 E and R cells were measured by RNA Sequencing (RNA-seq); and the results were further confirmed by real-time PCR. Reactive oxygen species (ROS), anoikis resistance, enzyme activity of aldehyde dehydrogenase 3 family, member A1 (ALDH3A1), and in vitro invasion assay were tested on both E and R cells. The deformability of HCT-8 E and R cells was measured by atomic force microscopy (AFM). To study the in vivo invasiveness of two cell types, athymic nude mice were intra-splenically injected with HCT-8 E or R cells and sacrificed after 9 weeks. Incidences of tumor development and metastasis were histologically evaluated and analyzed with Fisher's exact test.Results: Besides HCT-8, E-R transition on soft substrates was also seen in three other cancer cell lines (HCT116, SW480 colon and DU145 prostate cancer). The expression of some genes, such as ALDH3A1, TNS4, CLDN2, and AKR1B10, which are known to play important roles in cancer cell migration, invasion, proliferation and apoptosis, were increased in HCT-8 R cells. R cells also showed higher ALDH3A1 enzyme activity, higher ROS, higher anoikis resistance, and higher softness than E cells. More importantly, in vitro assay and in vivo animal models revealed that HCT-8 R cells were more invasive than E cells.Conclusions: Our comprehensive comparison of HCT-8 E and R cells revealed differences of molecular, phenotypical, and mechanical signatures between the two cell types. To our knowledge, this is the first study that explores the molecular mechanism of E-R transition, which may greatly increase our understanding of the mechanisms of cancer mechanical microenvironment and initiation of cancer metastasis.
KW - Cancer biomarkers
KW - In vitro cancer microenvironment
KW - Invasiveness
KW - Mechanotransduction
KW - Metastasis
KW - Polyacrylamide hydrogel
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U2 - 10.1186/1476-4598-13-131
DO - 10.1186/1476-4598-13-131
M3 - Article
C2 - 24884630
AN - SCOPUS:84903304046
SN - 1476-4598
VL - 13
JO - Molecular Cancer
JF - Molecular Cancer
IS - 1
M1 - 131
ER -