(Pro)renin receptor [(P)RR] is a receptor for renin and its precursor, prorenin. (P)RR plays physiological roles, not only in the renin-angiotensin system, but also in the activity of vacuolar H+-ATPase (V-ATPase)(Nguyen G. 2011). V-ATPase is responsible for the active transport of protons and maintains the acidity of intracellular compartments and the extracellular environment (Forgac, 2007). (P)RR is expressed in various types of cancer cells, including breast cancer and lung cancer. We have recently reported that suppression of (P)RR expression by (P)RR siRNA reduced cell proliferation and migration with suppressed autophagy in A549 lung cancer cells, suggesting that (P)RR plays a role in the cell proliferation and progression of cancer cells through the regulation of autophagy (Ohba et al., 2020). The aim of the present study is to clarify the effects of anti-cancer drugs on (P)RR expression, and the relation to apoptosis and autophagy in cancer cells. MCF-7 breast cancer cells and A549 lung cancer cells were treated with anti-cancer drugs, carboplatin or paclitaxel, and the expression of (P)RR, apoptosis markers and autophagy markers were studied by RT-qPCR, western blot analysis and immunocytochemistry. Expression levels of (P)RR mRNA were elevated 1.4-fold (p < 0.0001) at 48 hours in MCF-7 cells and 1.3-fold (p < 0.01) at 24 hours and 1.8-fold (p < 0.0001) at 48 hours in A549 cells by the treatment of carboplatin (100 µM)(n = 4). Expression levels of (P)RR mRNA were also elevated 1.3-fold (p < 0.01) at 48 hours in MCF-7 cells and 1.4-fold (p < 0.01) at 48 hours in A549 cells by the treatment of paclitaxel (10 nM) (n = 4). Western blot analysis showed that carboplatin increased expression levels of full-length (P)RR at 100 µM (1.2-fold, p < 0.05 compared to control) and soluble (P)RR at 50 and 100 µM (2.2-fold, p < 0.05 and 3.0-fold, p < 0.01 compared to control) in MCF-7 cells (n = 4). Carboplatin increased expression levels of soluble (P)RR at 100 µM (3.8-fold, p < 0.05, compared to control) in A549 cells (n = 4). Paclitaxel increased expression levels of soluble (P)RR at 10 nM in both MCF-7 and A549 cells (4.5-fold, p < 0.05, and 2.4-fold, p < 0.05, compared to control, respectively) (n = 4). Immunofluorescence staining of (P)RR was enhanced in both MCF-7 and A549 cells treated by carboplatin or paclitaxel. Apoptosis induction was shown by elevated BAX/BCL2 mRNA levels and increased active caspase3-positive cells. Moreover, autophagy induction was confirmed by increased levels of autophagy-associated mRNAs and LC3B-II proteins. The present study showed the expression of (P)RR mRNA and soluble (P)RR was increased by anti-cancer drugs with apoptosis and autophagy. Upregulated (P)RR and autophagy may act in protection of cancer cells from apoptosis.
Physiology 2021 (2021) Proc Physiol Soc 48, PC093
Poster Communications: Increased expression of (pro)renin receptor by anti-cancer drugs in cultured cancer cells: Relation to apoptosis and autophagy
Yurina Kashio-Yokota1, Shigemitsu Sato1, Takuo Hirose1, Tomoki Watanabe1, Akari Endo1, Fumihiko Watanabe1, Moe Endo1, Koji Ohba1, Takefumi Mori2, Kazuhiro Takahashi1
1 Tohoku University Graduate School of Medicine, Sendai, Japan 2 Tohoku Medical and Pharmaceutical University, Sendai, Japan
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Where applicable, experiments conform with Society ethical requirements.