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Synergistic suppression of human breast cancer cells by combination of plumbagin and zoledronic acid In vitro.

Qiao H, Wang TY, Yan W, Qin A, Fan QM, Han XG, Wang YG, Tang TT - Acta Pharmacol. Sin. (2015)

Bottom Line: Synergism was evaluated using Compusyn software, and the combination index (CI) and drug reduction index (DRI) values were determined.The DRI values also showed a synergistic effect between PL and ZA, with actual values of 5.52 and 3.59, respectively.Moreover, the combination of ZA and PL decreased the expression of Notch-1, cleaved PARP, Bcl-2 and Bcl-xl, and increased the expression of cleaved caspase-3, CDKN1A and ID1.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.

ABSTRACT

Aim: Zoledronic acid (ZA), a bisphosphonate, is currently used in combination with chemotherapeutic agents to suppress breast cancer cell proliferation or breast cancer-induced osteolysis. The aim of this study was to investigate the effects of ZA combined with a natural anticancer compound plumbagin (PL) against human breast cancer cells in vitro.

Methods: Human breast cancer MDA-MB-231SArfp cells were treated with ZA, PL or a combination of ZA and PL. The cell growth, apoptosis and migration were evaluated using CCK-8 assay, flow cytometry and transwell assay, respectively. The expression of apoptosis-related proteins was measured using real-time PCR and Western blotting. Synergism was evaluated using Compusyn software, and the combination index (CI) and drug reduction index (DRI) values were determined.

Results: PL or ZA alone caused mild cytotoxicity (the IC50 value at 24 h was 12.18 and above 100 μmol/L, respectively). However, the combination of ZA and PL caused a synergistic cytotoxicity (CI=0.26). The DRI values also showed a synergistic effect between PL and ZA, with actual values of 5.52 and 3.59, respectively. Furthermore, PL and ZA synergistically induced apoptosis and inhibited migration of the breast cancer cells. Moreover, the combination of ZA and PL decreased the expression of Notch-1, cleaved PARP, Bcl-2 and Bcl-xl, and increased the expression of cleaved caspase-3, CDKN1A and ID1. When the breast cancer cells were transfected with specific siRNA against Notch-1, the combination of ZA and PL markedly increased the expression of Bcl-2.

Conclusion: Combination of ZA and PL synergistically suppresses human breast cancer MDA-MB-231SArfp cells in vitro. PL can inhibit ZA-induced activation of the Notch-1 signaling pathway and subsequently reduce the expression of Bcl-2, thus potentiating cancer cell apoptosis.

No MeSH data available.


Related in: MedlinePlus

A schematic diagram of the proposed mechanisms of MDA-MB-231SArfp breast cancer cell apoptosis by synergistic treatment with plumbagin (PL) and zoledronic acid (ZA). According to the results of the present study, combined treatment with PL and ZA facilitates the expression of ID1, p21 (CDKN1A), cleaved caspase 3 while inhibiting the production of PARP and Notch-1, which contributes to decreased levels of Bcl-2 and results in the overall potentiation of apoptosis.
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fig8: A schematic diagram of the proposed mechanisms of MDA-MB-231SArfp breast cancer cell apoptosis by synergistic treatment with plumbagin (PL) and zoledronic acid (ZA). According to the results of the present study, combined treatment with PL and ZA facilitates the expression of ID1, p21 (CDKN1A), cleaved caspase 3 while inhibiting the production of PARP and Notch-1, which contributes to decreased levels of Bcl-2 and results in the overall potentiation of apoptosis.

Mentions: As shown in Figure 7D, Bcl-2 expression was lower in the PL-treatment group without siRNA silencing than in the PL-with-siRNA group. Additionally, treatment after siRNA silencing upregulated Bcl-2 expression compared with synergistic treatment alone, indicating that knocking down Notch-1 may result in the upregulation of Bcl-2 after combination treatment. The presence of Notch-1 promoted the downstream regulation of Bcl-2 and induced subsequent apoptosis enhancement after synergistic treatment. Thus, we demonstrated that synergistic suppression of Notch-1 signaling decreased the downstream, ZA-mediated expression of Bcl-2 and potentiated the apoptosis induced by the synergistic treatment (Figure 8).


Synergistic suppression of human breast cancer cells by combination of plumbagin and zoledronic acid In vitro.

Qiao H, Wang TY, Yan W, Qin A, Fan QM, Han XG, Wang YG, Tang TT - Acta Pharmacol. Sin. (2015)

A schematic diagram of the proposed mechanisms of MDA-MB-231SArfp breast cancer cell apoptosis by synergistic treatment with plumbagin (PL) and zoledronic acid (ZA). According to the results of the present study, combined treatment with PL and ZA facilitates the expression of ID1, p21 (CDKN1A), cleaved caspase 3 while inhibiting the production of PARP and Notch-1, which contributes to decreased levels of Bcl-2 and results in the overall potentiation of apoptosis.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4561967&req=5

fig8: A schematic diagram of the proposed mechanisms of MDA-MB-231SArfp breast cancer cell apoptosis by synergistic treatment with plumbagin (PL) and zoledronic acid (ZA). According to the results of the present study, combined treatment with PL and ZA facilitates the expression of ID1, p21 (CDKN1A), cleaved caspase 3 while inhibiting the production of PARP and Notch-1, which contributes to decreased levels of Bcl-2 and results in the overall potentiation of apoptosis.
Mentions: As shown in Figure 7D, Bcl-2 expression was lower in the PL-treatment group without siRNA silencing than in the PL-with-siRNA group. Additionally, treatment after siRNA silencing upregulated Bcl-2 expression compared with synergistic treatment alone, indicating that knocking down Notch-1 may result in the upregulation of Bcl-2 after combination treatment. The presence of Notch-1 promoted the downstream regulation of Bcl-2 and induced subsequent apoptosis enhancement after synergistic treatment. Thus, we demonstrated that synergistic suppression of Notch-1 signaling decreased the downstream, ZA-mediated expression of Bcl-2 and potentiated the apoptosis induced by the synergistic treatment (Figure 8).

Bottom Line: Synergism was evaluated using Compusyn software, and the combination index (CI) and drug reduction index (DRI) values were determined.The DRI values also showed a synergistic effect between PL and ZA, with actual values of 5.52 and 3.59, respectively.Moreover, the combination of ZA and PL decreased the expression of Notch-1, cleaved PARP, Bcl-2 and Bcl-xl, and increased the expression of cleaved caspase-3, CDKN1A and ID1.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.

ABSTRACT

Aim: Zoledronic acid (ZA), a bisphosphonate, is currently used in combination with chemotherapeutic agents to suppress breast cancer cell proliferation or breast cancer-induced osteolysis. The aim of this study was to investigate the effects of ZA combined with a natural anticancer compound plumbagin (PL) against human breast cancer cells in vitro.

Methods: Human breast cancer MDA-MB-231SArfp cells were treated with ZA, PL or a combination of ZA and PL. The cell growth, apoptosis and migration were evaluated using CCK-8 assay, flow cytometry and transwell assay, respectively. The expression of apoptosis-related proteins was measured using real-time PCR and Western blotting. Synergism was evaluated using Compusyn software, and the combination index (CI) and drug reduction index (DRI) values were determined.

Results: PL or ZA alone caused mild cytotoxicity (the IC50 value at 24 h was 12.18 and above 100 μmol/L, respectively). However, the combination of ZA and PL caused a synergistic cytotoxicity (CI=0.26). The DRI values also showed a synergistic effect between PL and ZA, with actual values of 5.52 and 3.59, respectively. Furthermore, PL and ZA synergistically induced apoptosis and inhibited migration of the breast cancer cells. Moreover, the combination of ZA and PL decreased the expression of Notch-1, cleaved PARP, Bcl-2 and Bcl-xl, and increased the expression of cleaved caspase-3, CDKN1A and ID1. When the breast cancer cells were transfected with specific siRNA against Notch-1, the combination of ZA and PL markedly increased the expression of Bcl-2.

Conclusion: Combination of ZA and PL synergistically suppresses human breast cancer MDA-MB-231SArfp cells in vitro. PL can inhibit ZA-induced activation of the Notch-1 signaling pathway and subsequently reduce the expression of Bcl-2, thus potentiating cancer cell apoptosis.

No MeSH data available.


Related in: MedlinePlus