Effects of Quinacrine on Expression of Hippo signaling Pathway Components (LATS1, LATS2, and YAP) in Human Breast Cancer Stem Cells

Abstract

Objective: The Hippo signaling pathway has important role in the pathogenesis of some tumors. Breast cancer is the most prevalent cancer among females in the world. In recent years, various articles referred to inhibiting effect of quinacrine, a derivative of 9-aminoacridine, on the growth of several types of cancer cells. In this study, we evaluated the effect of quinacrine on expression of LATS1, LATS2, and YAP genes of the Hippo signaling pathway and YAP level in human breast cancer stem cells (MDA-MB 231 cell line). This cell line of breast cancer expresses the triple negative characteristics.

Methods: MDA-MB 231 cells was treated with 0.5 µM of quinacrine for 3 days. The dose was selected using MTT assays. The expression of genes was quantified by Real-time PCR. The protein expression was performed by Western blotting. Significance of observations were checked by means of Mann-Whitney test using p.

Keywords: Cancer stem cells; Hippo signaling pathway; MDA-MB 231; quinacrine; triple negative breast cancer.

Figure 1

Alterations in the LATS1, LATS2 and YAP Genes Expression in MDA-MB 231 Cells Treated with 0.5 μM Quinacrine for 3 Days (Box-Whisker Plot). Graphs represent median for at least three replicates and the significance of differences were defined by mann whitney test (P < 0.05). Significant down regulation of YAP expression (fold: −2.00 and P = 0.014) is evident in treated cells but not change the expression of LATS1 and LATS2 is evident in treated cells (* P<0.05).

Figure 2

Agarose Gel Electrophoreses of PCR Products. A, PCR products of LATS1, YAP, and GAPDH genes (lines 6, 7, and 8 are negative controls), and B, PCR product of LAT2 gene (line 3 is negative control)

Figure 3

Expression Levels of YAP Protein. The expression of YAP oncoprotein was significantly less than control in MDA-MB 231 cells treated with 0.5 μM quinacrine for 3 days (fold: 61.0% ± 5.7 and P = 0.049). β-Actin was used as a housekeeping protein to normalize the expression (* P<0.05)