Trafficking of Full-Length and N-Terminally Truncated Cathepsin B in Human Colorectal Carcinoma Cells

Cathepsin B is an endo-lysosomal cysteine protease. However, its increased expression and altered localization to the extracellular space, to mitochondria, or to the nucleus has been linked to tumor progression. In the present study, we show enhanced levels of cathepsin B in adenocarcinoma tissue in comparison to adjacent normal colon. Additionally, cathepsin B was observed in the nuclear compartment of mucosal cells in adenocarcinoma tissue samples and in the nuclei of the colorectal carcinoma cell line HCT116. Accordingly, a distinct 40-kDa form of cathepsin B was detected in HCT116 cells, which is proposed to represent a specific form lacking the signal peptide and parts of the propeptide. Trafficking studies with an EGFP-tagged N-terminally truncated form, mimicking the 40-kDa form, demonstrated accumulation in aggresome-like inclusion bodies, while EGFP-tagged full-length cathepsin B revealed regular sorting to endo-lysosomes. We conclude that the identity of nuclear cathepsin B in colorectal adenocarcinoma (in situ) and in carcinoma cells (in vitro) cannot be attributed to either full-length or 40-kDa N-terminally truncated cathepsin B forms. Hence, future studies are needed to demonstrate which form/s of cathepsin B may be sorted to the nuclei of colorectal carcinoma cells, and whether redundant regulation of related cathepsin expression occurs.

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In Situ Localization of PPAR Gamma and Uncoupling Protein in Mouse Embryo Sections Using Digoxigenin-Labeled Riboprobes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162624 ◽

1996 ◽

Vol 54 ◽

pp. 32-33

Author(s):

C. Jennermann ◽

S. A. Kliewer ◽

D. C. Morris

Keyword(s):

Alkaline Phosphatase ◽

Room Temperature ◽

Uncoupling Protein ◽

Ppar Gamma ◽

Nuclear Hormone Receptor ◽

Full Length ◽

Northern Analysis ◽

Peroxisome Proliferator

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.

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Interleukin-8 as a growth factor for human colorectal carcinoma cells in vitro

Biochemical Society Transactions ◽

10.1042/bst025264s ◽

1997 ◽

Vol 25 (2) ◽

pp. 264S-264S ◽

Cited By ~ 21

Author(s):

ROBERT BREW ◽

JOHN S. ERIKSON ◽

DAVID C. WEST ◽

BRIAN F. FLANAGAN ◽

STEPHEN E. CHRISTMAS

Keyword(s):

Growth Factor ◽

Colorectal Carcinoma ◽

Carcinoma Cells ◽

Interleukin 8 ◽

Human Colorectal Carcinoma

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„IN VITRO“ AND „IN VIVO“ ANTITUMOR ACTIVITY OF PLATINUM(II) COMPLEXES WITH MALONIC ACID ON BREAST AND COLORECTAL CARCINOMA CELL LINES

10.46793/iccbi21.293f ◽

2021 ◽

Author(s):

Andjela Franich ◽

◽

Milica Dimitrijević Stojanović ◽

Snežana Rajković ◽

Marina Jovanović ◽

...

Keyword(s):

Antitumor Activity ◽

Colorectal Carcinoma ◽

Cell Lines ◽

Tumor Model ◽

Carcinoma Cells ◽

Spectroscopic Techniques ◽

Murine Cell Line ◽

In Vivo Antitumor Activity

Four Pt(II) complexes of the general formula [Pt(L)(5,6-epoxy-1,10-phen)], where L is anion of malonic (mal, Pt1), 2-methylmalonic (Me-mal, Pt2), 2,2-dimethylmalonic (Me2-mal, Pt3) or 1,1- cyclobutanedicarboxylic (CBDCA, Pt4) acid while 5,6-epoxy-1,10-phen is bidentately coordinated 5,6-epoxy-5,6-dihydro-1,10-phenanthroline were synthesized and characterized by elemental microanalysis, IR, UV-Vis and NMR (1H and 13C) spectroscopic techniques. In vitro anticancer activity of novel platinum(II) complexes have been investigated on human and murine cancer cell lines, as well as normal murine cell line by MTT assay. The obtained results indicate that studied platinum(II) complexes exhibited strong cytotoxic activity against murine breast carcinoma cells (4T1), human (HCT116) and murine (CT26) colorectal carcinoma cells. Complex Pt3 display stronger selectivity toward carcinoma cells in comparison to other tested platinum(II) complexes exhibiting beneficial antitumor activity mainly via the induction of apoptosis, as well as inhibition of cell proliferation and migration. Further study showed that Pt3 complex also carry significant in vivo antitumor activity in orthotopical 4T1 tumor model without detected liver, kidney, lung, and heart toxicity. All results imply that these novel platinum(II) complexes have a good anti-tumor effect on breast and colorectal cancer in vivo and in vitro and the affinity to become possible candidates for treatment in anticancer therapy.

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Grape Seed Extract Inhibits In vitro and In vivo Growth of Human Colorectal Carcinoma Cells

Clinical Cancer Research ◽

10.1158/1078-0432.ccr-06-1465 ◽

2006 ◽

Vol 12 (20) ◽

pp. 6194-6202 ◽

Cited By ~ 117

Author(s):

Manjinder Kaur ◽

Rana P. Singh ◽

Mallikarjuna Gu ◽

Rajesh Agarwal ◽

Chapla Agarwal

Keyword(s):

Colorectal Carcinoma ◽

Grape Seed Extract ◽

Grape Seed ◽

Seed Extract ◽

Carcinoma Cells ◽

Human Colorectal Carcinoma ◽

In Vivo Growth

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Role of in vitro exposure to TiO2 nanoparticles in human colorectal carcinoma cells cytotoxicity

ISEE Conference Abstracts ◽

10.1289/isee.2021.p-476 ◽

2021 ◽

Vol 2021 (1) ◽

Author(s):

Alfina Grasso ◽

Margherita Ferrante ◽

Massimo Libra ◽

Rossella Salemi ◽

Angelo Dimartino ◽

...

Keyword(s):

Colorectal Carcinoma ◽

Tio2 Nanoparticles ◽

Carcinoma Cells ◽

In Vitro Exposure ◽

Human Colorectal Carcinoma

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In Vitro Evaluation of Apoptotic Induction of Butyric Acid Derivatives in Colorectal Carcinoma Cells

Anticancer Research ◽

10.21873/anticanres.13528 ◽

2019 ◽

Vol 39 (7) ◽

pp. 3795-3801 ◽

Cited By ~ 6

Author(s):

LIJI PATTAYIL ◽

HARIKUMARAN-THAMPI BALAKRISHNAN-SARASWATHI

Keyword(s):

Colorectal Carcinoma ◽

Butyric Acid ◽

Carcinoma Cells ◽

In Vitro Evaluation ◽

Apoptotic Induction

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Flexicaulin A, An ent-Kaurane Diterpenoid, Activates p21 and Inhibits the Proliferation of Colorectal Carcinoma Cells through a Non-Apoptotic Mechanism

International Journal of Molecular Sciences ◽

10.3390/ijms20081917 ◽

2019 ◽

Vol 20 (8) ◽

pp. 1917 ◽

Cited By ~ 2

Author(s):

Yixuan Xia ◽

Chu Shing Lam ◽

Wanfei Li ◽

Md. Shahid Sarwar ◽

Kanglun Liu ◽

...

Keyword(s):

Colorectal Carcinoma ◽

Quantitative Polymerase Chain Reaction ◽

Carcinoma Cells ◽

Cell Viability Assay ◽

Human Carcinoma ◽

Viability Assay ◽

Human Colorectal Carcinoma ◽

Apoptotic Mechanism

Natural products, explicitly medicinal plants, are an important source of inspiration of antitumor drugs, because they contain astounding amounts of small molecules that possess diversifying chemical entities. For instance, Isodon (formerly Rabdosia), a genus of the Lamiaceae (formerly Labiatae) family, has been reported as a rich source of natural diterpenes. In the current study, we evaluated the in vitro anti-proliferative property of flexicaulin A (FA), an Isodon diterpenoid with an ent-kaurane structure, in human carcinoma cells, by means of cell viability assay, flow cytometric assessment, quantitative polymerase chain reaction array, Western blotting analysis, and staining experiments. Subsequently, we validated the in vivo antitumor efficacy of FA in a xenograft mouse model of colorectal carcinoma. From our experimental results, FA appears to be a potent antitumor molecule, since it significantly attenuated the proliferation of human colorectal carcinoma cells in vitro and restricted the growth of corresponsive xenograft tumors in vivo without causing any adverse effects. Regarding its molecular mechanism, FA considerably elevated the expression level of p21 and induced cell cycle arrest in the human colorectal carcinoma cells. While executing a non-apoptotic mechanism, we believe the antitumor potential of FA opens up new horizons for the therapy of colorectal malignancy.

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High Concentrations of Aspartame Induce Pro-Angiogenic Effects in Ovo and Cytotoxic Effects in HT-29 Human Colorectal Carcinoma Cells

Nutrients ◽

10.3390/nu12123600 ◽

2020 ◽

Vol 12 (12) ◽

pp. 3600 ◽

Cited By ~ 1

Author(s):

Anca Laura Maghiari ◽

Dorina Coricovac ◽

Iulia Andreea Pinzaru ◽

Ioana Gabriela Macașoi ◽

Iasmina Marcovici ◽

...

Keyword(s):

Colorectal Cancer ◽

Colorectal Carcinoma ◽

In Vitro Cytotoxicity ◽

Scientific Data ◽

Carcinoma Cells ◽

Artificial Sweetener ◽

In Ovo ◽

Human Colorectal Carcinoma ◽

Ht 29

Aspartame (ASP), an artificial sweetener abundantly consumed in recent years in an array of dietary products, has raised some concerns in terms of toxicity, and it was even suggested a link with the risk of carcinogenesis (colorectal cancer), though the present scientific data are rather inconclusive. This study aims at investigating the potential role of aspartame in colorectal cancer by suggesting two experimental approaches: (i) an in vitro cytotoxicity screening in HT-29 human colorectal carcinoma cells based on cell viability (Alamar blue assay), cell morphology and cell migration (scratch assay) assessment and (ii) an in ovo evaluation in terms of angiogenic and irritant potential by means of the chorioallantoic membrane method (CAM). The in vitro results showed a dose-dependent cytotoxic effect, with a significant decrease of viable cells at the highest concentrations tested (15, 30 and 50 mM) and morphological cellular changes. In ovo, aspartame (15 and 30 mM) proved to have a pro-angiogenic effect and a weak irritant potential at the vascular level. These data suggest new directions of research regarding aspartame’s role in colorectal cancer.

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