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Extracellular Caspase-8 Dependent Apoptosis on HeLa Cancer Cells and MRC-5 Normal Cells by ICD-85 (Venom Derived Peptides).

Zare-Mirakabadi A, Sarzaeem A - Iran J Cancer Prev (2012)

Bottom Line: The results show that Inhibitory Concentration 50% (IC50) value of ICD-85 for HeLa cells at 24 h was estimated and found to be 25.32±2.15 µg/mL.Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10 µg/mL did not significantly increase LDH release.Morphological changes in HeLa cells on treatment with ICD-85 compared with untreated HeLa cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, Iran.

ABSTRACT

Background: Our previous studies revealed an inhibitory effect of ICD-85 (venom derived peptides) on MDA-MB231 and HL-60 cell lines, through induction of apoptosis. The purpose of this study was to investigate apoptosis-induced mechanism on HeLa and MRC-5 cells by ICD-85 through activation of caspase-8.

Methods: Cell viability, cytosolic enzyme Lactate Dehydrogenase (LDH) and cell morphology were assessed under unexposed and ICD-85 exposed conditions.Caspase-8 activity was assayed by caspase-8 colorimetric assay Kit.

Results: The results show that Inhibitory Concentration 50% (IC50) value of ICD-85 for HeLa cells at 24 h was estimated and found to be 25.32±2.15 µg/mL. Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10 µg/mL did not significantly increase LDH release. Morphological changes in HeLa cells on treatment with ICD-85 compared with untreated HeLa cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies. However, when MRC-5 cells were exposed to ICD-85, no significant changes in cell morphology and LDH were observed at concentrations below 2.6×10µg/ml. Also, the apoptosis-induction mechanism by ICD-85 on HeLa cells was found through activation of caspase-8 and the activity of caspase-8 in HeLa cells was 1.5 folds more than its activity on MRC-5 cells.

Conclusion: Therefore, the apoptosis-induced mechanisms by ICD-85 are through activation of caspase-8 and concerning the least cytotoxic effect on MRC-5 cells, ICD-85 may be used as anticancer compound to inhibit growth of cancer cells.

No MeSH data available.


Related in: MedlinePlus

Effect of ICD-85 on morphology of HeLa and MRC-5 cells. HeLa and MRC-5 cells (1×106 cells/mL) were cultured in DMEM medium supplemented with 10% FBS and treated in the absence (control cells), or in the presence of ICD-85 at 5×10 μg/mL for 24 h at 37˚C. Morphological changes of treated cells were observed by inverted microscopy and compared with control cells. The arrows in figure B (40X) showed cell rounding (b) and granulation (a) in HeLa cancer cells treated with ICD-85 as compared to untreated HeLa cells (A, 40X). There are no significant morphological changes in MRC-5 normal cells treated with ICD-85 (D, 40X) as compared to untreated MRC-5 cells (C, 40X).
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f3-IJCP-05-194: Effect of ICD-85 on morphology of HeLa and MRC-5 cells. HeLa and MRC-5 cells (1×106 cells/mL) were cultured in DMEM medium supplemented with 10% FBS and treated in the absence (control cells), or in the presence of ICD-85 at 5×10 μg/mL for 24 h at 37˚C. Morphological changes of treated cells were observed by inverted microscopy and compared with control cells. The arrows in figure B (40X) showed cell rounding (b) and granulation (a) in HeLa cancer cells treated with ICD-85 as compared to untreated HeLa cells (A, 40X). There are no significant morphological changes in MRC-5 normal cells treated with ICD-85 (D, 40X) as compared to untreated MRC-5 cells (C, 40X).

Mentions: HeLa and MRC-5 cells were exposed to ICD-85 for 24 h and morphological changes were examined using invert microscopy. Figure 3-A and 3-C showing the morphology of control (unexposed to ICD-85) HeLa and MRC-5 cells respectively. While significant morphological changes in HeLa cancer cells were observed after ICD-85 treatment characterising the features of apoptosis such as cell rounding and granulation (Figure 3-B), no significant morphological alterations were found in MRC-5 cells treated with same concentration of ICD-85 (Figure 3-D).


Extracellular Caspase-8 Dependent Apoptosis on HeLa Cancer Cells and MRC-5 Normal Cells by ICD-85 (Venom Derived Peptides).

Zare-Mirakabadi A, Sarzaeem A - Iran J Cancer Prev (2012)

Effect of ICD-85 on morphology of HeLa and MRC-5 cells. HeLa and MRC-5 cells (1×106 cells/mL) were cultured in DMEM medium supplemented with 10% FBS and treated in the absence (control cells), or in the presence of ICD-85 at 5×10 μg/mL for 24 h at 37˚C. Morphological changes of treated cells were observed by inverted microscopy and compared with control cells. The arrows in figure B (40X) showed cell rounding (b) and granulation (a) in HeLa cancer cells treated with ICD-85 as compared to untreated HeLa cells (A, 40X). There are no significant morphological changes in MRC-5 normal cells treated with ICD-85 (D, 40X) as compared to untreated MRC-5 cells (C, 40X).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4209572&req=5

f3-IJCP-05-194: Effect of ICD-85 on morphology of HeLa and MRC-5 cells. HeLa and MRC-5 cells (1×106 cells/mL) were cultured in DMEM medium supplemented with 10% FBS and treated in the absence (control cells), or in the presence of ICD-85 at 5×10 μg/mL for 24 h at 37˚C. Morphological changes of treated cells were observed by inverted microscopy and compared with control cells. The arrows in figure B (40X) showed cell rounding (b) and granulation (a) in HeLa cancer cells treated with ICD-85 as compared to untreated HeLa cells (A, 40X). There are no significant morphological changes in MRC-5 normal cells treated with ICD-85 (D, 40X) as compared to untreated MRC-5 cells (C, 40X).
Mentions: HeLa and MRC-5 cells were exposed to ICD-85 for 24 h and morphological changes were examined using invert microscopy. Figure 3-A and 3-C showing the morphology of control (unexposed to ICD-85) HeLa and MRC-5 cells respectively. While significant morphological changes in HeLa cancer cells were observed after ICD-85 treatment characterising the features of apoptosis such as cell rounding and granulation (Figure 3-B), no significant morphological alterations were found in MRC-5 cells treated with same concentration of ICD-85 (Figure 3-D).

Bottom Line: The results show that Inhibitory Concentration 50% (IC50) value of ICD-85 for HeLa cells at 24 h was estimated and found to be 25.32±2.15 µg/mL.Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10 µg/mL did not significantly increase LDH release.Morphological changes in HeLa cells on treatment with ICD-85 compared with untreated HeLa cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, Iran.

ABSTRACT

Background: Our previous studies revealed an inhibitory effect of ICD-85 (venom derived peptides) on MDA-MB231 and HL-60 cell lines, through induction of apoptosis. The purpose of this study was to investigate apoptosis-induced mechanism on HeLa and MRC-5 cells by ICD-85 through activation of caspase-8.

Methods: Cell viability, cytosolic enzyme Lactate Dehydrogenase (LDH) and cell morphology were assessed under unexposed and ICD-85 exposed conditions.Caspase-8 activity was assayed by caspase-8 colorimetric assay Kit.

Results: The results show that Inhibitory Concentration 50% (IC50) value of ICD-85 for HeLa cells at 24 h was estimated and found to be 25.32±2.15 µg/mL. Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10 µg/mL did not significantly increase LDH release. Morphological changes in HeLa cells on treatment with ICD-85 compared with untreated HeLa cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies. However, when MRC-5 cells were exposed to ICD-85, no significant changes in cell morphology and LDH were observed at concentrations below 2.6×10µg/ml. Also, the apoptosis-induction mechanism by ICD-85 on HeLa cells was found through activation of caspase-8 and the activity of caspase-8 in HeLa cells was 1.5 folds more than its activity on MRC-5 cells.

Conclusion: Therefore, the apoptosis-induced mechanisms by ICD-85 are through activation of caspase-8 and concerning the least cytotoxic effect on MRC-5 cells, ICD-85 may be used as anticancer compound to inhibit growth of cancer cells.

No MeSH data available.


Related in: MedlinePlus