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Effects of Lipid Peroxidation-Derived Products on the Growth of Human Colorectal Cancer Cell Line HT-29.

Sakuma S, Sumi H, Kohda T, Arakawa Y, Fujimoto Y - J Clin Biochem Nutr (2009)

Bottom Line: However, to date the effects of lipid peroxidation-derived products that are formed from fat (especially free or esterified unsaturated fatty acids) on the initiation or progression of colorectal cancer have not been investigated extensively.Therefore, in the present study, we examined the effects of fatty acids, fatty acid hydroperoxides and aldehydes on the growth of human colorectal cancer cell line HT-29.At concentrations of 1 and 10 microM, linoleic, arachidonic and eicosapentaenoic acids, and 13-hydroperoxyoctadecadienoic and 15-hydroperoxyeicosapentaenoic acids had no significant effects on the growth of HT-29 cells. 4-Hydroxynonenal and 4-hydroxyhexenal had no significant effects on the growth of HT-29 cells up to 10 microM, whereas 4-oxononenal potently inhibited HT-29 cell growth (1-10 microM, 16-85% inhibition).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Physiological Chemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan.

ABSTRACT
Epidemiologic investigations indicate a close relationship between colorectal cancer and fat intake. However, to date the effects of lipid peroxidation-derived products that are formed from fat (especially free or esterified unsaturated fatty acids) on the initiation or progression of colorectal cancer have not been investigated extensively. Therefore, in the present study, we examined the effects of fatty acids, fatty acid hydroperoxides and aldehydes on the growth of human colorectal cancer cell line HT-29. At concentrations of 1 and 10 microM, linoleic, arachidonic and eicosapentaenoic acids, and 13-hydroperoxyoctadecadienoic and 15-hydroperoxyeicosapentaenoic acids had no significant effects on the growth of HT-29 cells. 4-Hydroxynonenal and 4-hydroxyhexenal had no significant effects on the growth of HT-29 cells up to 10 microM, whereas 4-oxononenal potently inhibited HT-29 cell growth (1-10 microM, 16-85% inhibition). Further experiments concerning DNA fragmentation, expression levels of Bax and Bcl-2 mRNA, expression levels of pro-caspase-3 and caspase-3 proteins, and activity of caspase-3 suggested that 4-oxononenal may increase the sensitivity of HT-29 cells to apoptosis through a decreased expression level of Bcl-2 and then increased formation of caspase-3 from pro-caspase-3.

No MeSH data available.


Related in: MedlinePlus

Effects of 4-ONE on the activity of caspase-3 (a), and on the protein expression levels of pro-caspase-3 and caspase-3 (b) in HT-29 cells. HT-29 cells were incubated with or without 10 µM 4-ONE for 12 h at 37°C in a humidified atmosphere of 5% CO2. Each bar indicates the mean of 3–4 experiments; vertical lines show SE. *p<0.01 vs control.
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Figure 5: Effects of 4-ONE on the activity of caspase-3 (a), and on the protein expression levels of pro-caspase-3 and caspase-3 (b) in HT-29 cells. HT-29 cells were incubated with or without 10 µM 4-ONE for 12 h at 37°C in a humidified atmosphere of 5% CO2. Each bar indicates the mean of 3–4 experiments; vertical lines show SE. *p<0.01 vs control.

Mentions: To clarify the mechanism by which 4-ONE inhibited HT-29 cell growth, its effects on the DNA fragmentation (Fig. 3), the expression levels of Bcl-2 and Bax mRNA (Fig. 4), the activity of caspase-3 and protein expression levels of pro-caspase-3 and caspase-3 (Fig. 5) were observed. Using the TUNEL method, DNA fragmentation was found as the fluorescence of small DNA particles in nuclei, when 4-ONE (10 µM) was added to HT-29 cells (Fig. 3a). The fragmentation of DNA in nuclei by incubation with 4-ONE (10 µM) was also visualized by agarose-gel electrophoresis of nuclear extracts (Fig. 3b); 4-ONE treatment resulted in small-sized DNA debris electrophoresed from the origin. Thus, 4-ONE showed DNA fragmentation in HT-29 cells in the same manner as TNF-α (positive control [7, 8]). As shown in Fig. 4, 4-ONE (10 µM) did not have any effect on Bax mRNA expression level, whereas it dramatically reduced the expression level of Bcl-2. Furthermore, 4-ONE (10 µM) enhanced the activity of caspase-3 in HT-29 cells (Fig. 5a). This might be partially due to a tentative increase in activated caspase-3 concomitant with a tentative decrease in pro-caspase-3 (Fig. 5b).


Effects of Lipid Peroxidation-Derived Products on the Growth of Human Colorectal Cancer Cell Line HT-29.

Sakuma S, Sumi H, Kohda T, Arakawa Y, Fujimoto Y - J Clin Biochem Nutr (2009)

Effects of 4-ONE on the activity of caspase-3 (a), and on the protein expression levels of pro-caspase-3 and caspase-3 (b) in HT-29 cells. HT-29 cells were incubated with or without 10 µM 4-ONE for 12 h at 37°C in a humidified atmosphere of 5% CO2. Each bar indicates the mean of 3–4 experiments; vertical lines show SE. *p<0.01 vs control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2735629&req=5

Figure 5: Effects of 4-ONE on the activity of caspase-3 (a), and on the protein expression levels of pro-caspase-3 and caspase-3 (b) in HT-29 cells. HT-29 cells were incubated with or without 10 µM 4-ONE for 12 h at 37°C in a humidified atmosphere of 5% CO2. Each bar indicates the mean of 3–4 experiments; vertical lines show SE. *p<0.01 vs control.
Mentions: To clarify the mechanism by which 4-ONE inhibited HT-29 cell growth, its effects on the DNA fragmentation (Fig. 3), the expression levels of Bcl-2 and Bax mRNA (Fig. 4), the activity of caspase-3 and protein expression levels of pro-caspase-3 and caspase-3 (Fig. 5) were observed. Using the TUNEL method, DNA fragmentation was found as the fluorescence of small DNA particles in nuclei, when 4-ONE (10 µM) was added to HT-29 cells (Fig. 3a). The fragmentation of DNA in nuclei by incubation with 4-ONE (10 µM) was also visualized by agarose-gel electrophoresis of nuclear extracts (Fig. 3b); 4-ONE treatment resulted in small-sized DNA debris electrophoresed from the origin. Thus, 4-ONE showed DNA fragmentation in HT-29 cells in the same manner as TNF-α (positive control [7, 8]). As shown in Fig. 4, 4-ONE (10 µM) did not have any effect on Bax mRNA expression level, whereas it dramatically reduced the expression level of Bcl-2. Furthermore, 4-ONE (10 µM) enhanced the activity of caspase-3 in HT-29 cells (Fig. 5a). This might be partially due to a tentative increase in activated caspase-3 concomitant with a tentative decrease in pro-caspase-3 (Fig. 5b).

Bottom Line: However, to date the effects of lipid peroxidation-derived products that are formed from fat (especially free or esterified unsaturated fatty acids) on the initiation or progression of colorectal cancer have not been investigated extensively.Therefore, in the present study, we examined the effects of fatty acids, fatty acid hydroperoxides and aldehydes on the growth of human colorectal cancer cell line HT-29.At concentrations of 1 and 10 microM, linoleic, arachidonic and eicosapentaenoic acids, and 13-hydroperoxyoctadecadienoic and 15-hydroperoxyeicosapentaenoic acids had no significant effects on the growth of HT-29 cells. 4-Hydroxynonenal and 4-hydroxyhexenal had no significant effects on the growth of HT-29 cells up to 10 microM, whereas 4-oxononenal potently inhibited HT-29 cell growth (1-10 microM, 16-85% inhibition).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Physiological Chemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan.

ABSTRACT
Epidemiologic investigations indicate a close relationship between colorectal cancer and fat intake. However, to date the effects of lipid peroxidation-derived products that are formed from fat (especially free or esterified unsaturated fatty acids) on the initiation or progression of colorectal cancer have not been investigated extensively. Therefore, in the present study, we examined the effects of fatty acids, fatty acid hydroperoxides and aldehydes on the growth of human colorectal cancer cell line HT-29. At concentrations of 1 and 10 microM, linoleic, arachidonic and eicosapentaenoic acids, and 13-hydroperoxyoctadecadienoic and 15-hydroperoxyeicosapentaenoic acids had no significant effects on the growth of HT-29 cells. 4-Hydroxynonenal and 4-hydroxyhexenal had no significant effects on the growth of HT-29 cells up to 10 microM, whereas 4-oxononenal potently inhibited HT-29 cell growth (1-10 microM, 16-85% inhibition). Further experiments concerning DNA fragmentation, expression levels of Bax and Bcl-2 mRNA, expression levels of pro-caspase-3 and caspase-3 proteins, and activity of caspase-3 suggested that 4-oxononenal may increase the sensitivity of HT-29 cells to apoptosis through a decreased expression level of Bcl-2 and then increased formation of caspase-3 from pro-caspase-3.

No MeSH data available.


Related in: MedlinePlus