Differential Expression of Tight Junctions and Cell Polarity Genes in Human Colon Cancer

Kourkoumpetis, Themistoklis; Royse, Kathryn E.; Chen, Liang; Ravishankar, Milan; Ittmann, Michael; El–Serag, Hashem B.; Jiao, Li

doi:10.14218/erhm.2017.00036

Cited by 3 publications

(4 citation statements)

References 24 publications

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“… 55 , 56 With regard to the expression levels of various tight and adherence junction proteins, we observed a lack in the expression of CLDN5 in our human cell line, whereas other markers are uniformly expressed. This could be a sign for a tumor characteristic of this line, 57 , 58 but this would contradict the microarray data. Therefore, after immortalization, the maintenance of the barrier integrity seems to be regulated by other claudin proteins as no reduced barrier integrity was observed in comparison to primary cell-based models.…”

Section: Discussionmentioning

confidence: 74%

Enhancing pre-clinical research with simplified intestinal cell line models

Fey,

Truschel,

Nehlsen

et al. 2024

J Tissue Eng

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Two-dimensional culture remains widely employed to determine the bioavailability of orally delivered drugs. To gain more knowledge about drug uptake mechanisms and risk assessment for the patient after oral drug admission, intestinal in vitro models demonstrating a closer similarity to the in vivo situation are needed. In particular, Caco-2 cell-based Transwell® models show advantages as they are reproducible, cost-efficient, and standardized. However, cellular complexity is impaired and cell function is strongly modified as important transporters in the apical membrane are missing. To overcome these limitations, primary organoid-based human small intestinal tissue models were developed recently but the application of these cultures in pre-clinical research still represents an enormous challenge, as culture setup is complex as well as time- and cost-intensive. To overcome these hurdles, we demonstrate the establishment of primary organoid-derived intestinal cell lines by immortalization. Besides exhibiting cellular diversity of the organoid, these immortalized cell lines enable a standardized and more cost-efficient culture. Further, our cell line-based Transwell®-like models display an organ-specific epithelial barrier integrity, ultrastructural features and representative transport functions. Altogether, our novel model systems are cost-efficient with close similarity to the in vivo situation, therefore favoring their use in bioavailability studies in the context of pre-clinical screenings.

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Section: Discussionmentioning

confidence: 74%

Enhancing pre-clinical research with simplified intestinal cell line models

Fey,

Truschel,

Nehlsen

et al. 2024

J Tissue Eng

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“…The characteristics of these study participants have been described. 7 We observed significant down-regulation of 29 genes in the autophagy pathway (p < 0.05) in colon cancer tissues compared to the normal colonic mucosa (Fig. 1, Supplemental Table S1).…”

Section: Resultsmentioning

confidence: 82%

“…The study population and research approaches were described previously. 7 Our study was conducted at the affiliated hospitals of Baylor College of Medicine (BCM) and involved six patients who were diagnosed with colon cancer and subsequently underwent total curative surgical excision during the years of 2007-2013. None received pre-operative neoadjuvant chemotherapy.…”

Section: Study Participantsmentioning

confidence: 99%

“…Thus, our analysis was restricted to the samples from five mucosal specimens with colon cancer and five biopsies from matched unrelated healthy controls. 7 We utilized the Qiagen RT 2 Profiler PCR Array (Valencia, CA, USA) to determine the expression of 84 key genes in the autophagy pathway (primer sequence is available upon request from Qiagen). The array includes genes encoding proteins for autophagic vacuole formation (AMBRA1 (NYW1), ATG12, ATG16L1, ATG4A, ATG4B, ATG4C, ATG4D, ATG5, ATG9A, ATG9B, BECN1, GABARAP, GABARAPL1, GABARAPL2, IRGM, MAP1LC3A, MAP1LC3B, RGS19, ULK1, WIPI1), vacuole targeting proteins (ATG4A, AT-G4B, ATG4C, ATG4D, GABARAP), transport proteins (ATG10, ATG16L1, ATG16L2, ATG3, ATG4A, ATG4B, ATG4C, ATG4D, ATG7, ATG9A, GABARAP, GABARAPL2, RAB24), autophagosome-lysosome linkage proteins (DRAM1, GABARAP, LAMP1, NPC1), ubiquitination proteins (ATG3, ATG7, HDAC6), and proteases (ATG4A, ATG4B, ATG4C, ATG4D).…”

Section: Rna Extraction and Gene Expression Analysismentioning

confidence: 99%

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Altered Expression of Autophagy-related Genes in Human Colon Cancer

2018

ERHM

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Background and objectives: Autophagy is a physiologic mechanism, which utilizes the self-digestion of cell organelles to promote cellular homeostasis, such as in the setting of dysfunctional cellular components, cellular stress or energy-deprived states. In vitro studies have pointed toward the key role of autophagy in colorectal cancer. However, in vivo studies from human colorectal cancer tissues are lacking. Methods: We collected tissue samples from six patients with colon cancer who received curative surgery at Baylor College of Medicine. We also obtained normal colonic mucosa biopsy from five unrelated polyp-free individuals who were matched to cases individually by age, sex, ethnicity, and colon segment. Total RNA was successfully extracted from fresh frozen tissue biopsies of five tumor tissues and five unrelated normal tissues. We tested the expression levels of 84 genes in a predefined autophagy pathway using the RT2 Profiler PCR array. We compared differences using Student’s t-test. The false-discovery rate was used for multiple testing adjustment. We also used the TCGA dataset to validate our findings. Results: We observed significant differential expression between colon cancer tissue and normal colon mucosa for 29 genes in the autophagy pathway (p < 0.05). After multiple testing adjustment, the expression of 17 genes was significantly down-regulated, with fold-change greater than 2 in colon cancer; these included ATG4A, ATG4C, ATG4D, and CTSS (q < 0.10). The down-regulation was observed in both early and late stage colon cancer. We observed the same down-regulation of multiple autophagy-related genes using the TCGA data. The ATG9B gene was the only statistically non-significantly up-regulated gene after multiple testing adjustment. Conclusions: This pilot study showed the down-regulation of multiple autophagy pathway genes in human colon cancer, corroborating the increasing clinical relevance of autophagy in human colorectal carcinogenesis. This preliminary finding should be validated in larger studies.

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Pathogenesis of Alcoholic Liver Disease

Kourkoumpetis

Sood

2019

Clinics in Liver Disease

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Differential Expression of Tight Junctions and Cell Polarity Genes in Human Colon Cancer

Cited by 3 publications

References 24 publications

Enhancing pre-clinical research with simplified intestinal cell line models

Enhancing pre-clinical research with simplified intestinal cell line models

Altered Expression of Autophagy-related Genes in Human Colon Cancer

Pathogenesis of Alcoholic Liver Disease

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