Analysis of proteomic changes induced upon cellular differentiation of the human intestinal cell line Caco‐2

Buhrke, Thorsten; Lengler, Imme; Lampen, Alfonso

doi:10.1111/j.1440-169x.2011.01258.x

Cited by 42 publications

(62 citation statements)

References 63 publications

(115 reference statements)

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“…In particular, the nucleus [20] and the plasma membrane [21] of those cells were compared to find proteins related with the differentiation process. Recently, a whole-cell approach was taken, and a comparison between the proteome of proliferating and differentiated cells revealed 53 proteins with differential regulation [6]. Interestingly, this study showed an upregulation in non-differentiated Caco-2 cells of proteins involved in cell growth or proliferation, and related to cancer, confirming the tumoral phenotype of these cells.…”

Section: Introductionmentioning

confidence: 53%

“…This cell line was established from a moderately well-differentiated human colon adenocarcinoma. When cultured in vitro over confluence under standard culture conditions, it spontaneously differentiates into a cell type with remarkable intestinal enterocyte-like features, including brush borders with microvilli on their apical side, tight junctions, and enterocytic hydrolase activities [3–6]. This differentiation process provides a valuable research tool, as undifferentiated Caco-2 cells resemble those found in tumor tissues, whereas the differentiated ones lose the tumorigenic phenotype and are similar to healthy enterocytes [7].…”

Section: Introductionmentioning

confidence: 99%

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Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

García-Lorenzo

Rodríguez-Piñeiro

Rodrı́guez-Berrocal

et al. 2012

IJMS

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Colorectal cancer is still a major health burden worldwide, and its diagnosis has not improved in recent years due to a lack of appropriate diagnostic serum markers. Aiming to find new diagnostic proteins, we applied the proteomic DIGE technology to analyze changes in the secretome before/after differentiation of the colon adenocarcinoma Caco-2 cell line, an accepted in vitro model to study colorectal tumorigenesis. When the secretomes from undifferentiated (tumor-like) and differentiated cells (resembling healthy enterocytes) were compared, we found 96 spots differentially expressed. After MS/MS analysis, 22 spots corresponding to 15 different proteins were identified. Principal component analysis demonstrated these 22 spots could serve as a discriminatory panel between the tumor-like and normal-like cells. Among the identified proteins, the translationally-controlled tumor protein (TCTP), the transforming growth factor-beta-induced protein ig-h3 (TGFβIp), and the Niemann-Pick disease type C2 protein (NPC2) are interesting candidates for future studies focused on their utility as serum biomarkers of colorectal cancer.

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Section: Introductionmentioning

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

García-Lorenzo

Rodríguez-Piñeiro

Rodrı́guez-Berrocal

et al. 2012

IJMS

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“…Following functional analysis, it was clear that a majority of identified proteins were associated with a decrease in (ALDOA, ARHGDIA, ATP5A1, CCT6A, GNB2L1, HSP90AB1, HSPA5, HSPA8, P4HB, PHB, and YWHAE) or regulation of (ENO1, HSPD1, NPM1, and PRDX2) cell death [41]. Many of the proteins involved in cell death regulation (i.e., ATP5A1, GNB2L1, HSPA5, HSPD1, NPM1, and PRDX2) were also relevant with respect to cell growth and proliferation [42], [43], [44]. Other proteins identified in this study (ANXA2, DES, FSCN1, LMNA, PKM2ENO1 and STAT5B) have also been characterized to regulate cell growth and proliferation [42], [43], [44].…”

Section: Discussionmentioning

confidence: 99%

“…Many of the proteins involved in cell death regulation (i.e., ATP5A1, GNB2L1, HSPA5, HSPD1, NPM1, and PRDX2) were also relevant with respect to cell growth and proliferation [42], [43], [44]. Other proteins identified in this study (ANXA2, DES, FSCN1, LMNA, PKM2ENO1 and STAT5B) have also been characterized to regulate cell growth and proliferation [42], [43], [44]. Only a few proteins, i.e., EGCG, HNRNPA2B1, and TPM1, have been characterized to induce cell death and/or decrease cell growth in response to various stimuli [45].…”

Section: Discussionmentioning

confidence: 99%

Cardiac-Oxidized Antigens Are Targets of Immune Recognition by Antibodies and Potential Molecular Determinants in Chagas Disease Pathogenesis

Dhiman

Zago

Nuñez³

et al. 2012

PLoS ONE

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Trypanosoma cruzi elicits reactive oxygen species (ROS) of inflammatory and mitochondrial origin in infected hosts. In this study, we examined ROS-induced oxidative modifications in the heart and determined whether the resultant oxidized cardiac proteins are targets of immune response and of pathological significance in Chagas disease. Heart biopsies from chagasic mice, rats and human patients exhibited, when compared to those from normal controls, a substantial increase in protein 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), carbonyl, and 3-nitrotyrosine (3-NT) adducts. To evaluate whether oxidized proteins gain antigenic properties, heart homogenates or isolated cardiomyocytes were oxidized in vitro and one- or two-dimensional gel electrophoresis (2D-GE)/Western blotting (WB) was performed to investigate the proteomic oxidative changes and recognition of oxidized proteins by sera antibodies in chagasic rodents (mice, rats) and human patients. Human cardiomyocytes exhibited LD50 sensitivity to 30 µM 4-HNE and 100 µM H2O2 at 6 h and 12 h, respectively. In vitro oxidation with 4-HNE or H2O2 resulted in a substantial increase in 4-HNE- and carbonyl-modified proteins that correlated with increased recognition of cardiac (cardiomyocytes) proteins by sera antibodies of chagasic rodents and human patients. 2D-GE/Western blotting followed by MALDI-TOF-MS/MS analysis to identify cardiac proteins that were oxidized and recognized by human chagasic sera yielded 82 unique proteins. We validated the 2D-GE results by enzyme-linked immunosorbent assay (ELISA) and WB and demonstrated that oxidation of recombinant titin enhanced its immunogenicity and recognition by sera antibodies from chagasic hosts (rats and humans). Treatment of infected rats with phenyl-α-tert-butyl nitrone (PBN, antioxidant) resulted in normalized immune detection of cardiac proteins associated with control of cardiac pathology and preservation of heart contractile function in chagasic rats. We conclude that ROS-induced, cardiac-oxidized antigens are targets of immune recognition by antibodies and molecular determinants for pathogenesis during Chagas disease.

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“…The human intestinal cancer cell line Caco-2 is a well-established model system to study cellular differentiation of human enterocytes since it differentiates spontaneously into polarized cells with morphological and biochemical features of small intestinal enterocytes [20]. Additionally, Caco-2 cells also differentiate when exposed to the physiologically relevant short-chain fatty acid, butyrate [19].…”

Section: Introductionmentioning

confidence: 99%

The Transition from Proliferation to Differentiation in Colorectal Cancer Is Regulated by the Calcium Activated Chloride Channel A1

et al. 2013

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Breaking the balance between proliferation and differentiation in animal cells can lead to cancer, but the mechanisms maintaining this balance remain largely undefined. The calcium activated chloride channel A1 (CLCA1) is a member of the calcium sensitive chloride conductance family of proteins and is expressed mainly in the colon, small intestine and appendix. We show that CLCA1 plays a functional role in differentiation and proliferation of Caco-2 cells and of intestinal tissue. Caco-2 cells spontaneously differentiate either in confluent culture or when treated with butyrate, a molecule present naturally in the diet. Here, we compared CLCA1 expressional levels between patients with and without colorectal cancer (CRC) and determined the functional role of CLCA1 in differentiation and proliferation of Caco-2 cells. We showed that: 1) CLCA1 and CLCA4 expression were down-regulated significantly in CRC patients; 2) CLCA1 expression was up-regulated in Caco-2 cells induced to differentiate by confluent culture or by treatment with sodium butyrate (NaBT); 3) Knockdown of CLCA1 with siRNA significantly inhibited cell differentiation and promoted cell proliferation in Caco-2 confluent cultures, and 4) In Caco-2 3D culture, suppression of CLCA1 significantly increased cell proliferation and compromised NaBT-induced inhibition of proliferation. In conclusion, CLCA1 may contribute to promoting spontaneous differentiation and reducing proliferation of Caco-2 cells and may be a target of NaBT-induced inhibition of proliferation and therefore a potential diagnostic marker for CRC prognosis.

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Analysis of proteomic changes induced upon cellular differentiation of the human intestinal cell line Caco‐2

Cited by 42 publications

References 63 publications

Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

Cardiac-Oxidized Antigens Are Targets of Immune Recognition by Antibodies and Potential Molecular Determinants in Chagas Disease Pathogenesis

The Transition from Proliferation to Differentiation in Colorectal Cancer Is Regulated by the Calcium Activated Chloride Channel A1

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