Cytotoxic and apoptotic activity of Canavalia seed extract in HT-29 human colon carcinoma cells

Introduction and Aim: Concanavalin the lectin from Canavalia spp. share high structural similarity among the same taxon and exhibit wide range of biological functions. The antiproliferative activity of lectins ConA (C. ensiformis), ConC (C. cathartica), ConG (C. gladiata) and ConM (C. rosea) were studied in human colon carcinoma. Materials and Methods: Human colon carcinoma HT-29 cells and human normal kidney HEK293T cells were examined. The cytotoxicity was evaluated by MTT assay. Cell apoptosis was analyzed by AO-EB (acridine orange-ethidium bromide dual staining) method. Cell cycle was investigated by flow cytometry (RNase-propidium iodide-based flow cytometry). The expression of cell signaling genes was analyzed by qRT-PCR. Results: All concanavalins significantly inhibited the proliferation of HT-29 cells in dose-dependent manner (IC50 values were ConG-10.45, ConA-14.86, ConC-33.34 and ConM-48.98 µg/mL respectively). ConA and ConG induced apoptotic morphology in HT-29 cells without affecting HEK293T cells. Similarly, both lectins increased the sub-G0/G1 proportions in HT-29 cells dose-dependently. ConG showed down-regulation of AKT1, ERK1/2 and mutant p53 in HT-29 cells. Conclusion: Concanavalin extract is shown to possess cytotoxic effect through inducing apoptosis, especially causing cell cycle arrest at G0/G1 phase in HT-29 cells.

Download Full-text

Src activity increases and Yes activity decreases during mitosis of human colon carcinoma cells.

Molecular and Cellular Biology ◽

10.1128/mcb.15.5.2374 ◽

1995 ◽

Vol 15 (5) ◽

pp. 2374-2382 ◽

Cited By ~ 40

Author(s):

J Park ◽

C A Cartwright

Keyword(s):

Cell Cycle ◽

Colon Carcinoma ◽

Human Colon ◽

T Antigen ◽

Carcinoma Cells ◽

Colon Carcinoma Cells ◽

Human Colon Carcinoma ◽

Polyomavirus Middle T Antigen ◽

Middle T Antigen ◽

Ht 29

Src and Yes protein-tyrosine kinase activities are elevated in malignant and premalignant tumors of the colon. To determine whether Src activity is elevated throughout the human colon carcinoma cell cycle as it is in polyomavirus middle T antigen- or F527 Src-transformed cells, and whether Yes activity, which is lower than that of Src in the carcinoma cells, is regulated differently, we measured their activities in cycling cells. We observed that the activities of both kinases were higher throughout all phases of the HT-29 colon carcinoma cell cycle than in corresponding phases of the fibroblast cycle. In addition, during mitosis of HT-29 cells, Src specific activity increased two- to threefold more, while Yes activity and abundance decreased threefold. The decreased steady-state protein levels of Yes during mitosis appeared to be due to both decreased synthesis and increased degradation of the protein. Inhibition of tyrosine but not serine/threonine phosphatases abolished the mitotic activation of Src. Mitotic Src was phosphorylated at novel serine and threonine sites and dephosphorylated at Tyr-527. Two cellular proteins (p160 and p180) were phosphorylated on tyrosine only during mitosis. Tyrosine phosphorylation of several other proteins decreased during mitosis. Thus, Src in HT-29 colon carcinoma cells, similar to Src complexed to polyomavirus middle T antigen or activated by mutation at Tyr-527, is highly active in all phases of the cell cycle. Moreover, Src activity further increases during mitosis, whereas Yes activity and abundance decrease. Thus, Src and Yes appear to be regulated differently during mitosis of HT-29 colon carcinoma cells.

Download Full-text

Silibinin upregulates the expression of cyclin-dependent kinase inhibitors and causes cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells

Oncogene ◽

10.1038/sj.onc.1207158 ◽

2003 ◽

Vol 22 (51) ◽

pp. 8271-8282 ◽

Cited By ~ 148

Author(s):

Chapla Agarwal ◽

Rana P Singh ◽

Sivanandhan Dhanalakshmi ◽

Anil K Tyagi ◽

Marianne Tecklenburg ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Arrest ◽

Colon Carcinoma ◽

Kinase Inhibitors ◽

Human Colon ◽

Cyclin Dependent Kinase ◽

Cycle Arrest ◽

Human Colon Carcinoma ◽

Ht 29

Download Full-text

Cladosporol a stimulates G1-phase arrest of the cell cycle by up-regulation of p21waf1/cip1 expression in human colon carcinoma HT-29 cells

Molecular Carcinogenesis ◽

10.1002/mc.20872 ◽

2011 ◽

Vol 52 (1) ◽

pp. 1-17 ◽

Cited By ~ 20

Author(s):

Diana Zurlo ◽

Cinzia Leone ◽

Gemma Assante ◽

Salvatore Salzano ◽

Giovanni Renzone ◽

...

Keyword(s):

Cell Cycle ◽

Colon Carcinoma ◽

Human Colon ◽

G1 Phase ◽

Human Colon Carcinoma ◽

Phase Arrest ◽

G1 Phase Arrest ◽

Ht 29

Download Full-text

Lidamycin induces marked G2 cell cycle arrest in human colon carcinoma HT-29 cells through activation of p38 MAPK pathway

Oncology Reports ◽

10.3892/or.17.3.597 ◽

2007 ◽

Author(s):

Xia Liu ◽

Chunjing Bian ◽

Kaihuan Ren ◽

Haixia Jin ◽

Baowei Li ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Arrest ◽

P38 Mapk ◽

Colon Carcinoma ◽

Mapk Pathway ◽

Human Colon ◽

Cycle Arrest ◽

Human Colon Carcinoma ◽

G2 Cell Cycle Arrest ◽

Ht 29

Download Full-text

Evidence of COX-2 independent induction of apoptosis and cell cycle block in human colon carcinoma cells after S- or R-ibuprofen treatment

European Journal of Pharmacology ◽

10.1016/j.ejphar.2006.04.030 ◽

2006 ◽

Vol 540 (1-3) ◽

pp. 24-33 ◽

Cited By ~ 32

Author(s):

Astrid Janssen ◽

Thorsten J. Maier ◽

Susanne Schiffmann ◽

Ovidiu Coste ◽

Maic Seegel ◽

...

Keyword(s):

Cell Cycle ◽

Colon Carcinoma ◽

Human Colon ◽

Carcinoma Cells ◽

Colon Carcinoma Cells ◽

Human Colon Carcinoma ◽

Induction Of Apoptosis ◽

Cox 2 ◽

Cell Cycle Block

Download Full-text

Protein kinase C and casein kinase II activities in two human colon carcinoma cell lines, HT-29 and CaCo-2: Possible correlation with differentiation

Bioscience Reports ◽

10.1007/bf01117245 ◽

1990 ◽

Vol 10 (3) ◽

pp. 293-299 ◽

Cited By ~ 6

Author(s):

Ewa Rydell ◽

Karl-Eric Magnusson ◽

Anita Sjö ◽

Krister Axelsson

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Colon Carcinoma ◽

Carcinoma Cell ◽

Human Colon ◽

Human Colon Carcinoma Cell ◽

Carcinoma Cell Lines ◽

Kinase C ◽

Human Colon Carcinoma ◽

Ht 29

Protein kinase C (PK-C) and casein kinase II (CK-II) activities were studied in two human colon carcinoma cell lines (HT-29 and CaCO-2) undergoing differentiation in vitro resulting, in small-intestine-like cells. CaCo-2 cells, when grown under standard conditions, appear to undergo spontaneous differentiation. In these cells PK-C and CK-II activities were determined on day 5, 10 and 15. No significant differences in activities were seen either in PK-C or CK-II activity. HT-29 cells, when grown in glucose-free medium can be stimulated to undergo differentiation which is completed within 20 days. PK-C and CK-II activities were determined after 5, 10, 15, 20 and 25 days, respectively. PK-C activity rose from 7.9±3.5 pmole32P/mg protein/min at day 5 to 37.5±14.8 pmole32P/mg protein/min at day 20. After 25 days the activity was reduced to 20.0±7.8 pmole32P/mg protein/min. CK-II activity did not change significantly during day 5 to 20, but on day 25 there was a significant decrease in CK-II activity from 94.9±6.4 pmole32P/mg protein/min (day 20) to 62.6±3.9 pmole32P/mg protein/min (day 25) p=0.003. The results in this study indicate a role for PK-C and CK-II in cell growth and differentiation.

Download Full-text