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Apoptosis Induction in Primary Human Colorectal Cancer Cell Lines and Retarded Tumor Growth in SCID Mice by Sulforaphane.

Chen MJ, Tang WY, Hsu CW, Tsai YT, Wu JF, Lin CW, Cheng YM, Hsu YC - Evid Based Complement Alternat Med (2011)

Bottom Line: In addition, treated cells showed nuclear apoptotic morphology that coincided with an activation of caspase-3, and loss of mitochondrial membrane potential (ΔΨm).Incubations at higher SFN doses (12.5 and 25 μM) resulted in cleavage of procaspase-3 and elevated caspase-2, -3, -8, and -9 activity, suggesting that the induction of apoptosis and the sulforaphane-induced mitosis delay at the lower dose are independently regulated.Daily SFN s.c. injections (400 micromol/kg/d for 3 weeks) in severe combined immunodeficient mice with primary human CRC (CP1 to CP5) s.c. tumors resulted in a decrease of mean tumor weight by 70% compared with vehicle-treated controls.

View Article: PubMed Central - PubMed

Affiliation: Division of Traumatology, Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan.

ABSTRACT
We have investigated the anticancer effects of the dietary isothiocyanate sulforaphane (SFN) on colorectal cancer (CRC), using primary cancer cells lines isolated from five Taiwanese colorectal cancer patients as the model for colorectal cancer. SFN-treated cells accumulated in metaphase (SFN 6.25 μM) and subG1 (SFN 12.5 and 25 μM) as determined by flow cytometry. In addition, treated cells showed nuclear apoptotic morphology that coincided with an activation of caspase-3, and loss of mitochondrial membrane potential (ΔΨm). Incubations at higher SFN doses (12.5 and 25 μM) resulted in cleavage of procaspase-3 and elevated caspase-2, -3, -8, and -9 activity, suggesting that the induction of apoptosis and the sulforaphane-induced mitosis delay at the lower dose are independently regulated. Daily SFN s.c. injections (400 micromol/kg/d for 3 weeks) in severe combined immunodeficient mice with primary human CRC (CP1 to CP5) s.c. tumors resulted in a decrease of mean tumor weight by 70% compared with vehicle-treated controls. Our findings suggest that, in addition to the known effects on cancer prevention, sulforaphane may have antitumor activity in established colorectal cancer.

No MeSH data available.


Related in: MedlinePlus

Arrest of cell cycle progression at G2/M and subG0/G1 in response to SFN treatment. (a) SFN-induced G2/M (SFN 6.25 μM) and G0/G1 (SFN 12.5 and 25 μM) arrest in primary CRC cell lines. The distribution of the cell cycle of CRC cell lines was assessed by flow cytometry after staining with propidium iodide (PI). (b) Results were expressed as a percentage of subG0/G1. All data were reported as the means (±SEM) of at least three separate experiments. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus SFN 0 μM group, while the symbol on the bar denotes the difference which is statistically significant at P < 0.05 as compared to the SFN 6.25 (#) or 12.5 μM (&).
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fig2: Arrest of cell cycle progression at G2/M and subG0/G1 in response to SFN treatment. (a) SFN-induced G2/M (SFN 6.25 μM) and G0/G1 (SFN 12.5 and 25 μM) arrest in primary CRC cell lines. The distribution of the cell cycle of CRC cell lines was assessed by flow cytometry after staining with propidium iodide (PI). (b) Results were expressed as a percentage of subG0/G1. All data were reported as the means (±SEM) of at least three separate experiments. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus SFN 0 μM group, while the symbol on the bar denotes the difference which is statistically significant at P < 0.05 as compared to the SFN 6.25 (#) or 12.5 μM (&).

Mentions: Cell cycle distribution of SFN-treated CRC cell lines was analyzed by flow cytometry, aiming to determine whether the inhibitory effect was due to cell cycle arrest and apoptosis. Before being processed and analyzed, both kinds of cells were exposed to SFN for a total of 24 h. As shown in Figure 2(a), the CRC cells exposed to SFN 6.25 μM showed G2/M arrest, but SFN 12.5 and 25 μM showed increase in the number of cells in the subG0/G1 phase, as compared with that of the untreated cells. These results revealed that SFN could hold up CRC cell line proliferation via caused cell cycle arrest in the G2/M phase and a significant increase in the proportion of cells in the subG0/G1 phase. The results in Figure 2(b) suggest that the five CRC cell lines have accumulated subG0/G1 phase (y = 11.602x − 12.985  R2 = 0.9139) following the SFN treatment for 24 hours. The observations could imply that the primary colorectal cancer cells have undergone apoptosis.


Apoptosis Induction in Primary Human Colorectal Cancer Cell Lines and Retarded Tumor Growth in SCID Mice by Sulforaphane.

Chen MJ, Tang WY, Hsu CW, Tsai YT, Wu JF, Lin CW, Cheng YM, Hsu YC - Evid Based Complement Alternat Med (2011)

Arrest of cell cycle progression at G2/M and subG0/G1 in response to SFN treatment. (a) SFN-induced G2/M (SFN 6.25 μM) and G0/G1 (SFN 12.5 and 25 μM) arrest in primary CRC cell lines. The distribution of the cell cycle of CRC cell lines was assessed by flow cytometry after staining with propidium iodide (PI). (b) Results were expressed as a percentage of subG0/G1. All data were reported as the means (±SEM) of at least three separate experiments. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus SFN 0 μM group, while the symbol on the bar denotes the difference which is statistically significant at P < 0.05 as compared to the SFN 6.25 (#) or 12.5 μM (&).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
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fig2: Arrest of cell cycle progression at G2/M and subG0/G1 in response to SFN treatment. (a) SFN-induced G2/M (SFN 6.25 μM) and G0/G1 (SFN 12.5 and 25 μM) arrest in primary CRC cell lines. The distribution of the cell cycle of CRC cell lines was assessed by flow cytometry after staining with propidium iodide (PI). (b) Results were expressed as a percentage of subG0/G1. All data were reported as the means (±SEM) of at least three separate experiments. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus SFN 0 μM group, while the symbol on the bar denotes the difference which is statistically significant at P < 0.05 as compared to the SFN 6.25 (#) or 12.5 μM (&).
Mentions: Cell cycle distribution of SFN-treated CRC cell lines was analyzed by flow cytometry, aiming to determine whether the inhibitory effect was due to cell cycle arrest and apoptosis. Before being processed and analyzed, both kinds of cells were exposed to SFN for a total of 24 h. As shown in Figure 2(a), the CRC cells exposed to SFN 6.25 μM showed G2/M arrest, but SFN 12.5 and 25 μM showed increase in the number of cells in the subG0/G1 phase, as compared with that of the untreated cells. These results revealed that SFN could hold up CRC cell line proliferation via caused cell cycle arrest in the G2/M phase and a significant increase in the proportion of cells in the subG0/G1 phase. The results in Figure 2(b) suggest that the five CRC cell lines have accumulated subG0/G1 phase (y = 11.602x − 12.985  R2 = 0.9139) following the SFN treatment for 24 hours. The observations could imply that the primary colorectal cancer cells have undergone apoptosis.

Bottom Line: In addition, treated cells showed nuclear apoptotic morphology that coincided with an activation of caspase-3, and loss of mitochondrial membrane potential (ΔΨm).Incubations at higher SFN doses (12.5 and 25 μM) resulted in cleavage of procaspase-3 and elevated caspase-2, -3, -8, and -9 activity, suggesting that the induction of apoptosis and the sulforaphane-induced mitosis delay at the lower dose are independently regulated.Daily SFN s.c. injections (400 micromol/kg/d for 3 weeks) in severe combined immunodeficient mice with primary human CRC (CP1 to CP5) s.c. tumors resulted in a decrease of mean tumor weight by 70% compared with vehicle-treated controls.

View Article: PubMed Central - PubMed

Affiliation: Division of Traumatology, Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan.

ABSTRACT
We have investigated the anticancer effects of the dietary isothiocyanate sulforaphane (SFN) on colorectal cancer (CRC), using primary cancer cells lines isolated from five Taiwanese colorectal cancer patients as the model for colorectal cancer. SFN-treated cells accumulated in metaphase (SFN 6.25 μM) and subG1 (SFN 12.5 and 25 μM) as determined by flow cytometry. In addition, treated cells showed nuclear apoptotic morphology that coincided with an activation of caspase-3, and loss of mitochondrial membrane potential (ΔΨm). Incubations at higher SFN doses (12.5 and 25 μM) resulted in cleavage of procaspase-3 and elevated caspase-2, -3, -8, and -9 activity, suggesting that the induction of apoptosis and the sulforaphane-induced mitosis delay at the lower dose are independently regulated. Daily SFN s.c. injections (400 micromol/kg/d for 3 weeks) in severe combined immunodeficient mice with primary human CRC (CP1 to CP5) s.c. tumors resulted in a decrease of mean tumor weight by 70% compared with vehicle-treated controls. Our findings suggest that, in addition to the known effects on cancer prevention, sulforaphane may have antitumor activity in established colorectal cancer.

No MeSH data available.


Related in: MedlinePlus