Research on LPA and Hippo-YAP signaling pathway in promoting invasion and metastasis of TNBC cell MDA-MB-231 / 医学研究生学报

ABSTRACT

Objective Triple negative breast cancer(TNBC), a special breast cancer subtype, is lack of effective target therapy.The article aimed to investigate the role of lysophosphatidic acid (LPA) and Hippo Yes-associated protein (Hippo-YAP) signaling pathway in TNBC invasion and metastasis and the mechanisms.Methods The specific small interfering RNA (siRNA) of YAP was synthetized in vitro, and was transfected into MDA-MB-231 cells using liposome transfection.The experiment was divided into YAP-siRNA group, positive control group and blank control group.Each group is provided with 2 parallel holes.Evaluation was made on the effects of each group on Hippo-YAP, the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group, YAP content, the capacity of invasion and metastasis after transfection ([0.035±0.005], [2.200±1.000], [3.500±0.800]) significantly decreased compared with positive control group([0.343±0.012], [27.600±5.100], [22.300±5.000]) and blank control group([0.384±0.017], [26.500±4.800], [22.350±6.000]) (P<0.05).YAP expression levels at 60 min, 120 min, and 240 min in experiment group significantly decreased compared with positive control group and blank control group (P<0.05).YAP relative expression levels of 10, 20, 50 μmol/Lwere significantly lower than those of positive control group and blank control group (P<0.05).After respective interference of C3 transferase and Y27623, significant difference was found in the pYAP mRNA contents of experiment group([0.255±0.052], [0.326±0.017]), blank control group([0.048±0.032], [0.534±0.017]) and positive control group([0.052±0.021], [0.528±0.024])(P<0.05).The expression levels of YAP mNA and AREG mNA significantly increased in experiment group([0.176±0.032], [0.263±0.008]) compared with blank control group([0.043±0.013], [0.263±0.008]) and positive control group([0.049±0.025], [0.057±0.043])(P<0.05).Conclusion LPA induces breast cancer invasion and metastasis, which is YAP-dependent, time-dependent and concentration-dependent.LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.