Cytotoxic effects of hydroalcoholic extract of Cuscuta chinensis on PC3 and MCF7 cancer cell lines

Abstract

Objective: Chemoprevention of cancer by application of natural phytochemical compounds has been used to prevent, delay or suppress cancer progression. Cuscuta chinensis a traditional Iranian medicinal herb, has biological properties including anticancer, anti-aging, immuno-stimulatory and antioxidant effects. In this study, anti-proliferative effects of hydroalcoholic extract of C. chinensis on prostate (PC3) and breast (MCF7) cancer cell lines were investigated.

Materials and methods: In the current study, we investigated treatment of PC3 cells with different concentrations of C. chinensis (0, 100, 200, 300, 400, and 500 µg/ml) for 24 and 48 hr; also, MCF7 cells were treated with various concentrations (0-600 µg/ml) of C. chinensis for 48 and 72 hr and cell viability was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. mRNA expression of BCL2 Associated X (Bax), B-cell lymphoma 2 (Bcl2), Cysteine-aspartic proteases (Caspase3) and Phosphatase and tensin homolog (PTEN) were analyzed by quantitative real-time PCR. Annexin V/PI staining and lactate dehydrogenase (LDH) cytotoxicity assay were used to detect apoptosis.

Results: C. chinensis decreased PC3 and MCF7 cells viability in a dose- and time-dependent manner (p<0.01 to p<0.001). The gene expression of BAX/Bcl2 ratio, Caspase3 and PTEN increased in C. chinensis-treated cells compared to the control group. C. chinensis induced apoptosis (p<0.001) and LDH activity (p<0.01 to p<0.001).

Conclusion: Our findings suggest that C. chinensis extract is able to inhibit proliferation and induce apoptosis in PC3 and MCF7 cell lines. Therefore, C. chinensis extract exerts antitumor activity against cancer cells.

Keywords: Apoptosis; Breast cancer; Chemoprevention; Cuscuta. Chinensis; Prostate cancer.

Figure 1

Effect of C. chinensis on MCF7 (a) and PC3 (b) cells viability. PC3 cells were treated with 0, 200, 300, and 400 µg/ml of C. chinensis and for 24 and 48 hr and MCF7 cells were treated with 0, 100, 200, and 300 µg/ml of C. chinensis for 48 and 72 hr and their viability were examined by MTT assay. Data are reported as the mean±SEM (n=8). **p<0.01 and ***p<0.001 compared to the control

Figure 2

Effect of C. chinensis on BAX/Bcl2, CASPASE3 and PTEN expression. Relative expression genes in MCF7 (a and b) and PC3 (c and d) cells was determined by real time PCR. PC3 cells were treated with 0, 200, 300, and 400 µg/ml of C. chinensis for 24 and 48 hr and MCF7 cells were treated with 0, 100, 200, and 300 µg/ml of C. chinensis for 48 and 72 hr. The 2^−ΔΔCT method was used for data analysis. Data are reported as the mean±SEM. *p<0.05, **p<0.01, and ***p<0.001 show significant differences compared to the control

Figure 3

Effect of C. chinensis on MCF7 (a) and PC3 (b) on LDH activity. PC3 cells were treated with 0, 200, 300, and 400 µg/ml of C. chinensis for 24 and 48 hr and MCF7 cells were treated with 0, 100, 200, and 300 µg/ml of C. chinensis for 48 and 72 hr. LDH activity was assessed in cell lysate using colorimetric kit. C.chinensis increased LDH activity. Data are expressed as the mean±SEM. **p<0.01 and ***p<0.001 show significant differences versus the control. LDH: Lactate dehydrogenase

Figure 4

Representative flow cytometric analysis of treatment of MCF7 (a and b) for 48 and 72 hr with respectively 400 and 200 μM C. chinensis and PC3 (c and d) with 300 and 200 μM C. chinensis for respectively 24 and 48 hr. Data are expressed as mean±SEM. ***p<0.001 shows significant differences vs untreated cells