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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

SFN activates procaspase-3 degradation in five CRC cell lines. The cells were treated with SFN (0, 6.25, 12.5, and 25 μM) for 24 hours, and then Western blot analysis was performed for procaspase-3. (a) Representative blot from 3 independent experiments. (b) Quantification of band intensities by Li-COR near infrared imaging system. (c) The caspase-2, -3, -8, -9 activity was analyzed by ApoAlert Caspase assay plates. SFN induces the caspase activity of five CRC cell lines. (d) Quantification of DNA fragmentation by measuring the fluorescence intensities by flow cytometry. The data showed that DNA fragmentation levels were significantly elevated in cells incubated with SFN incubation for 24 hours. All data were reported as the means (±SEM) of five CRC cell lines. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus 0 μM control group.
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fig6: SFN activates procaspase-3 degradation in five CRC cell lines. The cells were treated with SFN (0, 6.25, 12.5, and 25 μM) for 24 hours, and then Western blot analysis was performed for procaspase-3. (a) Representative blot from 3 independent experiments. (b) Quantification of band intensities by Li-COR near infrared imaging system. (c) The caspase-2, -3, -8, -9 activity was analyzed by ApoAlert Caspase assay plates. SFN induces the caspase activity of five CRC cell lines. (d) Quantification of DNA fragmentation by measuring the fluorescence intensities by flow cytometry. The data showed that DNA fragmentation levels were significantly elevated in cells incubated with SFN incubation for 24 hours. All data were reported as the means (±SEM) of five CRC cell lines. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus 0 μM control group.

Mentions: Figure 6(a) shows the immunoblotting of cellular proteins from five CRC cell lines treated with SFN showing decrease of procaspase-3 after SFN incubation. Quantification of procaspase-3, done by measuring the relative band intensities, showed that procaspase-3 levels were significantly lower in cells incubated with SFN (Figure 6(b)). The results indicated that SFN induced caspase-3 activity via cleaved procaspase-3 and apoptosis after SFN incubation. As shown in Figure 6(c), the SFN elevated caspase-2, -3, -8, -9 activities in five CRC cell lines that have been decreased with caspase-specific inhibitors. The results summarized in Figure 6 indicate that the increased levels of caspase activity may play an important role in SFN-induced apoptosis in CRC cell lines.


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)

SFN activates procaspase-3 degradation in five CRC cell lines. The cells were treated with SFN (0, 6.25, 12.5, and 25 μM) for 24 hours, and then Western blot analysis was performed for procaspase-3. (a) Representative blot from 3 independent experiments. (b) Quantification of band intensities by Li-COR near infrared imaging system. (c) The caspase-2, -3, -8, -9 activity was analyzed by ApoAlert Caspase assay plates. SFN induces the caspase activity of five CRC cell lines. (d) Quantification of DNA fragmentation by measuring the fluorescence intensities by flow cytometry. The data showed that DNA fragmentation levels were significantly elevated in cells incubated with SFN incubation for 24 hours. All data were reported as the means (±SEM) of five CRC cell lines. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus 0 μM control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig6: SFN activates procaspase-3 degradation in five CRC cell lines. The cells were treated with SFN (0, 6.25, 12.5, and 25 μM) for 24 hours, and then Western blot analysis was performed for procaspase-3. (a) Representative blot from 3 independent experiments. (b) Quantification of band intensities by Li-COR near infrared imaging system. (c) The caspase-2, -3, -8, -9 activity was analyzed by ApoAlert Caspase assay plates. SFN induces the caspase activity of five CRC cell lines. (d) Quantification of DNA fragmentation by measuring the fluorescence intensities by flow cytometry. The data showed that DNA fragmentation levels were significantly elevated in cells incubated with SFN incubation for 24 hours. All data were reported as the means (±SEM) of five CRC cell lines. Statistical analysis used the t-test, with the significant differences determined at the level of *P < 0.05 versus 0 μM control group.
Mentions: Figure 6(a) shows the immunoblotting of cellular proteins from five CRC cell lines treated with SFN showing decrease of procaspase-3 after SFN incubation. Quantification of procaspase-3, done by measuring the relative band intensities, showed that procaspase-3 levels were significantly lower in cells incubated with SFN (Figure 6(b)). The results indicated that SFN induced caspase-3 activity via cleaved procaspase-3 and apoptosis after SFN incubation. As shown in Figure 6(c), the SFN elevated caspase-2, -3, -8, -9 activities in five CRC cell lines that have been decreased with caspase-specific inhibitors. The results summarized in Figure 6 indicate that the increased levels of caspase activity may play an important role in SFN-induced apoptosis in CRC cell lines.

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