Zoledronic acid inhibits TSC2-null cell tumor growth via RhoA/YAP signaling pathway in mouse models of lymphangioleiomyomatosis

Abstract

Background: This study is to investigate the effects of zoledronic acid (ZA) on TSC2-null cell proliferation and on the tumor progression and recurrence in mouse models of lymphangioleiomyomatosis (LAM). Methods:Subcutaneous mouse models and LAM mouse models were established. Immunohistochemistry and immunofluorescence were performed to detect the protein expression levels. TUNEL assay was conducted to detect cell apoptosis. Immunoprecipitation was carried out to determine the interaction between proteins.Results: ZA prevented the growth of TSC2-null cells both in culture and in LAM mouse models. Compared with rapamycin, ZA more effectively promoted the apoptosis of TSC2-null cells. Moreover, combined with the rapamycin, ZA effectively suppressed the tumor recurrence after drug withdrawal and ZA inhibited the activity of GTPase RhoA by decreasing protein geranylgeranylation, resulting in changes of Yap nucleus translocation. Conclusion: ZA promotes cell apoptosis in TSC2-null cells through the RhoA/YAP signaling pathway. ZA may be used for the clinical treatment of LAM. Additional file 2: Fig. S1. ZA induces autophagy in TSC2-null cells. (A) Phosphorylated S6 and Yap expression detected by immunoblotting analysis in TSC2-null cells treated with ZA (25, 50, and 100 μM, respectively) for 24 h. (B) LC3 expression was detected by immunoblotting analysis in TSC2-null cells treated with ZA (25, 50, 75, and 100 μM, respectively) for 24 h. (C) Localization of LC3 and LAMP1 was analyzed by immunofluorescence in TSC2-null cells treated with ZA (25 and 50 μM, respectively) alone and RAPA (20 nM) for 24 h. (D) Localization of LC3 and LAMP1 was analyzed by immunofluorescence in TSC2-null cells treated with ZA (50 μM), RAPA (20 nM), and combination of ZA (50 μM) and GGPP (20 μM) for 24 h.The experiment was performed for three times. All data were presented as mean ± SEM. Compared with the placebo, * P < 0.05.

Actin Alpha 2 (ACTA2) Downregulation Inhibits Neural Stem Cell Migration through Rho GTPase Activation

“…We focused our attention on the RhoA signaling pathway as the increased level of RhoA was higher than the decreased level of Rac1. In addition, previous studies have proven that mediation of RhoA regulates proliferation and differentiation of mesenchymal stem cells [26][27][28][29], adipose-derived stem cells (ADSCs) [30], muscle stem cells [31], and a bulk of cancer cells [18,32,33].…”

“…For NSC passage, neurospheres were collected by centrifugation at the speed of 300 rpm, dissociated in StemPro Accutase Cell Dissociation Reagent (Gibco, Grand Island, NY), and grown in enrichment medium as described above. Y27632 was purchased from Sigma-Aldrich (St. Louis, MO, USA), and the working concentration was 30 μM as previously reported [18].…”

Gamma-Delta CAR-T Cells Show CAR-Directed and Independent Activity Against Leukemia

“…For all in vivo experiments, 8-15-weeks-old NOD-SCID-IL-2Rγ-(NSG) female mice, purchased from the Jackson laboratories, were used. Mice were tail-vein injected with 1 × 10 6 Nalm6 cells for leukemia inoculation, followed by intravenous (IV) injections of effector cells with or without intraperitoneal (IP) injections of zoledronate (20).…”