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Induction of apoptosis in HCT-116 colon cancer cells by polysaccharide of Larimichthys crocea swim bladder.

Suo H, Song JL, Zhou Y, Liu Z, Yi R, Zhu K, Xie J, Zhao X - Oncol Lett (2014)

Bottom Line: In particular, 400 μg/ml PLCSB significantly (P<0.05) induced apoptosis, which was demonstrated by 4,6-diamidino-2-phenylindole staining and flow cytometry analysis.To elucidate the mechanisms underlying the anticancer effect of PLCSB in HCT-116 cancer cells, the expression of apoptosis and metastasis-associated genes was analyzed by reverse transcription-polymerase chain reaction and western blot analysis.A total of 400 μg/ml PLCSB significantly induced apoptosis in HCT-116 cells (P<0.05) via the upregulation Bax, p53, p21, apoptotic protease activating factor 1, caspase-3, -8, and -9, as well as Fas and the downregulation of B-cell lymphoma 2 (Bcl-2), Bcl-extra large and Fas ligand (L).

View Article: PubMed Central - PubMed

Affiliation: College of Food Science, Southwest University, Chongqing 400715, P.R. China.

ABSTRACT

Larimichthys crocea swim bladder is a traditional food and medicine widely used in China. The in vitro anticancer effects of polysaccharide of L. crocea swim bladder (PLCSB) in HCT-116 human colon cancer cells was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. At concentrations ranging between 0 and 800 μg/ml PLCSB, cancer cell viability was decreased by PLCSB in a concentration-dependent manner. In particular, 400 μg/ml PLCSB significantly (P<0.05) induced apoptosis, which was demonstrated by 4,6-diamidino-2-phenylindole staining and flow cytometry analysis. To elucidate the mechanisms underlying the anticancer effect of PLCSB in HCT-116 cancer cells, the expression of apoptosis and metastasis-associated genes was analyzed by reverse transcription-polymerase chain reaction and western blot analysis. A total of 400 μg/ml PLCSB significantly induced apoptosis in HCT-116 cells (P<0.05) via the upregulation Bax, p53, p21, apoptotic protease activating factor 1, caspase-3, -8, and -9, as well as Fas and the downregulation of B-cell lymphoma 2 (Bcl-2), Bcl-extra large and Fas ligand (L). The results of this study demonstrated that PLCSB exhibits an anticancer effect on HCT-116 colon cancer cells, in vitro.

No MeSH data available.


Related in: MedlinePlus

Exposure of HCT-116 human colon carcinoma cells to PLCSB induces apoptosis. (A) The appearance of apoptotic bodies in HCT-116 cells treated with PLCSB was determined by 4,6-diamidino-2-phenylindole assay. (B) Treatment with HCT-116 increased the number of apoptotic cells as measured by flow cytometry. The profile represents an increased sub-G1 population (apoptotic cells). PLCSB, polysaccharide of Larimichthys crocea swim bladder.
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f2-ol-09-02-0972: Exposure of HCT-116 human colon carcinoma cells to PLCSB induces apoptosis. (A) The appearance of apoptotic bodies in HCT-116 cells treated with PLCSB was determined by 4,6-diamidino-2-phenylindole assay. (B) Treatment with HCT-116 increased the number of apoptotic cells as measured by flow cytometry. The profile represents an increased sub-G1 population (apoptotic cells). PLCSB, polysaccharide of Larimichthys crocea swim bladder.

Mentions: To determine a possible mechanism underlying the growth inhibitory activity of PLCSB in HCT-116 cancer cells, the induction of apoptosis was analyzed. The extent of chromatin condensation was determined by fluorescence microscopy of the cells stained with the DNA-binding fluorescent dye DAPI and flow cytometric analysis. While the untreated HCT-116 cells exhibited nuclei with homogeneous chromatin distribution, treatment with the PLCSB induced chromatin condensation and nuclear fragmentation, which indicated the presence of apoptotic cells (Fig. 2A). Chromatin condensation and the formation of apoptotic bodies, which are two hallmarks of apoptosis, were observed in cells cultured with 200 and 400 μg/ml PLCSB. By contrast, the level of chromatin condensation was low in the cells treated with 100 μg/ml PLCSB. Flow cytometric analyses revealed that treatment with PLCSB promoted apoptosis of the HCT-116 cells, compared with the untreated control cancer cells. This conclusion is based on the significant accumulation of cells with a sub-G1 DNA content (Fig. 2B). The induction of apoptosis was almost negligible (2.7%) in untreated control cancer cells; however, cancer cells treated with 400 μg/ml PLCSB exhibited a higher level of apoptosis (37.2%) than cells treated with 100 μg/ml (13.7%) and 200 μg/ml (20.5%) PLCSB.


Induction of apoptosis in HCT-116 colon cancer cells by polysaccharide of Larimichthys crocea swim bladder.

Suo H, Song JL, Zhou Y, Liu Z, Yi R, Zhu K, Xie J, Zhao X - Oncol Lett (2014)

Exposure of HCT-116 human colon carcinoma cells to PLCSB induces apoptosis. (A) The appearance of apoptotic bodies in HCT-116 cells treated with PLCSB was determined by 4,6-diamidino-2-phenylindole assay. (B) Treatment with HCT-116 increased the number of apoptotic cells as measured by flow cytometry. The profile represents an increased sub-G1 population (apoptotic cells). PLCSB, polysaccharide of Larimichthys crocea swim bladder.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4301533&req=5

f2-ol-09-02-0972: Exposure of HCT-116 human colon carcinoma cells to PLCSB induces apoptosis. (A) The appearance of apoptotic bodies in HCT-116 cells treated with PLCSB was determined by 4,6-diamidino-2-phenylindole assay. (B) Treatment with HCT-116 increased the number of apoptotic cells as measured by flow cytometry. The profile represents an increased sub-G1 population (apoptotic cells). PLCSB, polysaccharide of Larimichthys crocea swim bladder.
Mentions: To determine a possible mechanism underlying the growth inhibitory activity of PLCSB in HCT-116 cancer cells, the induction of apoptosis was analyzed. The extent of chromatin condensation was determined by fluorescence microscopy of the cells stained with the DNA-binding fluorescent dye DAPI and flow cytometric analysis. While the untreated HCT-116 cells exhibited nuclei with homogeneous chromatin distribution, treatment with the PLCSB induced chromatin condensation and nuclear fragmentation, which indicated the presence of apoptotic cells (Fig. 2A). Chromatin condensation and the formation of apoptotic bodies, which are two hallmarks of apoptosis, were observed in cells cultured with 200 and 400 μg/ml PLCSB. By contrast, the level of chromatin condensation was low in the cells treated with 100 μg/ml PLCSB. Flow cytometric analyses revealed that treatment with PLCSB promoted apoptosis of the HCT-116 cells, compared with the untreated control cancer cells. This conclusion is based on the significant accumulation of cells with a sub-G1 DNA content (Fig. 2B). The induction of apoptosis was almost negligible (2.7%) in untreated control cancer cells; however, cancer cells treated with 400 μg/ml PLCSB exhibited a higher level of apoptosis (37.2%) than cells treated with 100 μg/ml (13.7%) and 200 μg/ml (20.5%) PLCSB.

Bottom Line: In particular, 400 μg/ml PLCSB significantly (P<0.05) induced apoptosis, which was demonstrated by 4,6-diamidino-2-phenylindole staining and flow cytometry analysis.To elucidate the mechanisms underlying the anticancer effect of PLCSB in HCT-116 cancer cells, the expression of apoptosis and metastasis-associated genes was analyzed by reverse transcription-polymerase chain reaction and western blot analysis.A total of 400 μg/ml PLCSB significantly induced apoptosis in HCT-116 cells (P<0.05) via the upregulation Bax, p53, p21, apoptotic protease activating factor 1, caspase-3, -8, and -9, as well as Fas and the downregulation of B-cell lymphoma 2 (Bcl-2), Bcl-extra large and Fas ligand (L).

View Article: PubMed Central - PubMed

Affiliation: College of Food Science, Southwest University, Chongqing 400715, P.R. China.

ABSTRACT

Larimichthys crocea swim bladder is a traditional food and medicine widely used in China. The in vitro anticancer effects of polysaccharide of L. crocea swim bladder (PLCSB) in HCT-116 human colon cancer cells was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. At concentrations ranging between 0 and 800 μg/ml PLCSB, cancer cell viability was decreased by PLCSB in a concentration-dependent manner. In particular, 400 μg/ml PLCSB significantly (P<0.05) induced apoptosis, which was demonstrated by 4,6-diamidino-2-phenylindole staining and flow cytometry analysis. To elucidate the mechanisms underlying the anticancer effect of PLCSB in HCT-116 cancer cells, the expression of apoptosis and metastasis-associated genes was analyzed by reverse transcription-polymerase chain reaction and western blot analysis. A total of 400 μg/ml PLCSB significantly induced apoptosis in HCT-116 cells (P<0.05) via the upregulation Bax, p53, p21, apoptotic protease activating factor 1, caspase-3, -8, and -9, as well as Fas and the downregulation of B-cell lymphoma 2 (Bcl-2), Bcl-extra large and Fas ligand (L). The results of this study demonstrated that PLCSB exhibits an anticancer effect on HCT-116 colon cancer cells, in vitro.

No MeSH data available.


Related in: MedlinePlus