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The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene.

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Presentation on theme: "The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene."— Presentation transcript:

1 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 1. Histological study of normal and treated breast cancer tissues. A) Normal breast tissue. B) Breast tumor induced by LA7. C) Breast tissue from low dose of TPHE treatment group. D) Breast tissue from high dose of TPHE treatment group E) Breast tissue from tamoxifen treated group. H and E staining of the tumor tissues shows morphological differences between normal breast tissues and cancerous breast tissue. The treatment of the tumor with TPHE shows reorganisation if the tumor part to the normal tissues. Magnification 40x. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

2 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 2. Induced apoptosis of breast cancer cells as determined by the TUNEL assay. A) Control tumor tissue. B) Breast tissue from low dose of TPHE treatment group C) Breast tissue from high dose of TPHE treatment group. D) Breast tissue from tamoxifen treated group. Breast cancer tissue sections from the control and treatment group were subjected to the TUNEL assay. Tissue from the control group did not shows any significant changes in the color of the tissues While tissue from the high dose TPHE and tamoxifen treated groups showed a very high number of apoptotic cells. The low dose TPHE treated group did not show significant apoptotic activity. Magnification 40x. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

3 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 3. Proliferation markers (Ki67 and PCNA) decreased after Treatment. A) Ki67 Tumor control B) Ki67 low dose TPHE treatment C) Ki67 high dose TPHE treatment D) Ki67 tamoxifen treatment E & F) PCNA tumor control. Immunohistochemical staining with Ki-67 and PCNA shows the reduction of the Ki67 and PCNA markers after treatment with TPHE and tamoxifen in low and high dose treatment (G). The data represent the means ± SEM of three independent experiments. *p<0.05 compared with the control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

4 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 4. Immunohistochemistry staining of Bax, Bcl-2, P53 and Caspase 3 proteins. A) Tumor control B) Low dose TPHE treatment C) High dose TPHE treatment D) tamoxifen treatment. The analysis of Bax and Bcl-2 shows an up-regulation of Bax, p53 and caspase 3 and down-regulation of Bcl-2 which revealed the induction of apoptosis through mitochondrial pathway. Magnification 40x. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

5 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 5. Immunohistochemistry staining of p21 and p27 A) Tumor control B) Low dose treatment C) High dose treatment D) Tamoxifen treatment. Cell cycle arrest was confirmed by high expression of p21 and p27 proteins in treatment groups when compare to control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

6 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 6. Chemical structure of the compound HDZC. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

7 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 7. Flow chart of bioassay-guided isolation of HDZC from the hexane extract of T. polycephalum leaves. Each fraction was subjected to MTT assay for the evaluation of its suppressive effect on MCF7 cells. IC50 values represent the means ± SEM of three independent experiments. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

8 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 8. Cell cycle analysis. Cell cycle analysis of MCF7 cells treated with (A) vehicle 0.1% DMSO (control) for 48 h and HDZC (IC50 concentration) for (B) 12, (C) 24 and (D) 48 h. (E) The representative bar chart illustrated the significant G1 cell cycle arrest after 12, 24 and 48 h. The data represent the means ± SEM of three independent experiments. *p<0.05 compared with the control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

9 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 9. Effect of HDZC on apoptosis rates in MCF7 cells. MCF7 cells was determined via quadrant statistics. Cell were treated with (A) vehicle 0.1% DMSO (control) for 48 h and HDZC (IC50 concentration) for (B) 12, (C) 24 and (D) 48 h. (E) The representative bar chart illustrates the proportion of viable, early apoptotic, late apoptotic and necrotic MCF7 cells. The data represent the means ± SEM of three independent experiments. *p<0.05 compared to the control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

10 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 10. Effect of HDZC on the activation of caspase 9 and caspase 7. The caspase 7 and 9 on MCF7 cells was determined using a bioluminescent analysis. MCF7 cells were treated with the vehicle 0.1% DMSO (control) for 48 h and HDZC (IC50 concentration) for 3, 6, 12, 24 and 48 h. Caspase 9 and caspase 7 were significantly activated after 12, 24 and 48 h treatment. The data represent the means ± SEM of three independent experiments. *p<0.05 compared with the control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

11 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 11. Q-PCR analysis of apoptosis- and cell cycle-associated molecules, viz. Bcl-2, Bax, caspase 9, caspase 7, p21 and p27. MCF7 cells were exposed to HDZC at IC50 concentration for 12, 24 and 48 h. The mRNA expression of genes was represented after normalization with β-actin housekeeping gene. The data represent the means ± SEM of three independent experiments. *p<0.05 compared with the control group. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

12 The Chemopreventive Effect of <b><i>Tanacetum Polycephalum</i></b> Against LA7-Induced Breast Cancer in Rats and the Apoptotic Effect of a Cytotoxic Sesquiterpene Lactone in MCF7 Cells: A Bioassay-Guided Approach Cell Physiol Biochem 2015;36: DOI: / Fig. 12. Western blot analysis of apoptosis- and cell cycle-associated molecules, viz. Bcl-2, Bax, caspase 9, caspase 7, p21 and p27. MCF7 cells were exposed to HDZC at IC50 concentration for 12, 24 and 48 h. β-actin was used as a positive control. The data represent the means ± SEM of three independent experiments. © 2015 S. Karger AG, Basel - CC BY-NC 3.0

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