Extracts from Curcuma zedoaria Inhibit Proliferation of Human Breast Cancer Cell MDA-MB-231 In Vitro

Abstract

Objective. To evaluate the effect of petroleum ether extracts of Curcuma zedoaria on the proliferation of human triple negative breast cancer cell line MDA-MB-231. Methods. The reagents were isolated from Curcuma zedoaria by petroleum ether fraction. It was assayed by CCK8 for MDA-MB-231 cellular viability with various concentrations and days, cell cycle analyses, Western Blot analysis, and Realtime Reverse Transcriptase PCR analyses for chemokines molecules including E-cadherin, and E-selectin, and adhesion molecules including CCR7, SLC, SDF-1, and CXCR4. Epirubicin was used as control in the study. Results. MDA-MB-231 cells were inhibited by petroleum ether extracts of Curcuma zedoaria (P < 0.05), and the inhibition rate was dependent on concentrations and time. Petroleum ether extracts of Curcuma zedoaria as well as Epirubicin produce a significant G0/G1 cell cycle arrest. The level of expression of proteins E-cadherin and E-cadherin mRNA was significantly increased, while proteins SDF-1, CCR7, and CXCR4 mRNA were decreased after being incubated with petroleum ether extracts of Curcuma zedoaria at the concentrations of 300 μ g/mL than control (P < 0.05). The differences were that the protein CXCR4 mRNA expression level was higher than vehicle. Conclusions. MDA-MB-231 cells were inhibited by petroleum ether extracts of Curcuma zedoaria.

Figure 1

Curcuma zedoaria of the plant and the plant root sample.

Figure 2

The inhibition rate of the various concentrations of epirubicin (a) and petroleum ether extracts of Curcuma zedoaria (b) were determined by CCK8 assays. The values were reported as %. (c) The inhibition rate of different days was determined by CCK8 assay as described compared with vehicle. Values are average of triplicate experiment and are represented as mean ± SD.

Figure 3

Synchronized MDA-MB-231 cells were treated with petroleum ether extracts of Curcuma zedoaria or Epirubicin for 24 hours, and the fraction of cells in each phase of cell cycle was evaluated by flow cytometry.

Figure 4

Effect of petroleum ether extracts of Curcuma zedoaria (300 μg/mL) and Epirubicin (1 μg/mL) on protein E-cadherin, E-selectin, CCR7, SLC, SDF-1, and CXCR4 expression after 24 hours. Bars represent mean ± SD of three individual experiments. *P < 0.05. Beta-actin was used as a loading control.

Figure 5

The level of adhesion molecules E-cadherin and E-selectin mRNA expression (a) and chemokines molecules CCR7, SLC, SDF-1, and CXCR4 (b) after 24 hours in different groups by using RT-PCR analysis. *P < 0.01 compared with control.

Figure 6

The inhibition rate of the different extracts of Curcuma zedoaria was determined by CCK8 assay as described compared with Epirubicin. The values were reported as %.