Flavonoids kaempferol and quercetin are nuclear receptor 4A1 (NR4A1, Nur77) ligands and inhibit rhabdomyosarcoma cell and tumor growth

Abstract Background Flavonoids exhibit both chemopreventive and chemotherapeutic activity for multiple tumor types, however, their mechanisms of action are not well defined. Based on some of their functional and gene modifying activities as anticancer agents, we hypothesized that kaempferol and quercetin were nuclear receptor 4A1 (NR4A1, Nur77) ligands and confirmed that both compounds directly bound NR4A1 with KD values of 3.1 and 0.93 μM, respectively. Methods The activities of kaempferol and quercetin were determined in direct binding to NR4A1 protein and in NR4A1-dependent transactivation assays in Rh30 and Rh41 rhabdomyosarcoma (RMS) cells. Flavonoid-dependent effects as inhibitors of cell growth, survival and invasion were determined in XTT and Boyden chamber assays respectively and changes in protein levels were determined by western blots. Tumor growth inhibition studies were carried out in athymic nude mice bearing Rh30 cells as xenografts. Results Kaempferol and quercetin bind NR4A1 protein and inhibit NR4A1-dependent transactivation in RMS cells. NR4A1 also regulates RMS cell growth, survival, mTOR signaling and invasion. The pro-oncogenic PAX3-FOXO1 and G9a genes are also regulated by NR4A1 and, these pathways and genes are all inhibited by kaempferol and quercetin. Moreover, at a dose of 50 mg/kg/d kaempferol and quercetin inhibited tumor growth in an athymic nude mouse xenograft model bearing Rh30 cells. Conclusion These results demonstrate the clinical potential for repurposing kaempferol and quercetin for clinical applications as precision medicine for treating RMS patients that express NR4A1 in order to increase the efficacy and decrease dosages of currently used cytotoxic drugs.

Download Full-text

Flavonoids Kaempferol and Quercetin are Nuclear Receptor 4A1 (NR4A1, Nur77) Ligands and inhibit Rhabdomyosarcoma Cell and Tumor Growth

10.21203/rs.3.rs-638373/v1 ◽

2021 ◽

Author(s):

Rupesh Shrestha ◽

Kumaravel Mohankumar ◽

Greg Martin ◽

Amanuel Hailemariam ◽

Syng-ook Lee ◽

...

Keyword(s):

Cell Growth ◽

Tumor Growth ◽

Nuclear Receptor ◽

Anticancer Agents ◽

Xenograft Model ◽

Boyden Chamber ◽

Tumor Growth Inhibition ◽

Western Blots ◽

Multiple Tumor ◽

Mouse Xenograft Model

Abstract BackgroundFlavonoid’s exhibit both chemopreventive and chemotherapeutic activity for multiple tumor types, however, their mechanisms of action are not well defined. Based on some of their functional and gene modifying activities as anticancer agents, we hypothesized that kaempferol and quercetin were nuclear receptor 4A1 (NR4A1, Nur77) ligands and confirmed that both compounds directly bound NR4A1 with KD values of 3.1 and 0.93 µM, respectively. MethodsThe activities of kaempferol and quercetin were determined in direct binding to NR4A1 and in NR4A1-dependent transactivation assays in Rh30 and Rh41 rhabdomyosarcoma (RMS) cells, flavonoid-dependent effects as inhibitors of cell growth, survival and invasion were determined in XTT, Annexin V and Boyden chamber assays respectively and changes in protein levels were determined by western blots. Tumor growth inhibition studies were carried out in athymic nude mice bearing Rh30 cells. ResultsKaempferol and quercetin bind NR4A1 protein and inhibit NR4A1-dependent transactivation in RMS cells. NR4A1 also regulates RMS cell growth, survival, and invasion and pro-oncogenic PAX3-FOX01 and G9a gene expression, mTOR signaling and gene products and, these pathways and genes are inhibited by kaempferol and quercetin. Moreover, at a dose of 50 mg/kg/d kaempferol and quercetin inhibited tumor growth in athymic nude mouse xenograft model bearing Rh30 cells. ConclusionThese results demonstrate the clinical potential for repurposing these flavonoids for clinical applications as precision medicine for treating RMS patients that express NR4A1 in order to increase the efficacy and decrease dosages of currently used cytotoxic drugs.

Download Full-text

Doxorubicin Embedded into Nanofibrillated Bacterial Cellulose (NFBC) Produces a Promising Therapeutic Outcome for Peritoneally Metastatic Gastric Cancer in Mice Models via Intraperitoneal Direct Injection

Nanomaterials ◽

10.3390/nano11071697 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1697

Author(s):

Hidenori Ando ◽

Takashi Mochizuki ◽

Amr S. Abu Lila ◽

Shunsuke Akagi ◽

Kenji Tajima ◽

...

Keyword(s):

Gastric Cancer ◽

Drug Delivery ◽

Tumor Growth ◽

Bacterial Cellulose ◽

Anticancer Agents ◽

Intraperitoneal Administration ◽

In Vitro Release ◽

Xenograft Model ◽

Tumor Growth Inhibition

Natural materials such as bacterial cellulose are gaining interest for their use as drug-delivery vehicles. Herein, the utility of nanofibrillated bacterial cellulose (NFBC), which is produced by culturing a cellulose-producing bacterium (Gluconacetobacter intermedius NEDO-01) in a medium supplemented with carboxymethylcellulose (CMC) that is referred to as CM-NFBC, is described. Recently, we demonstrated that intraperitoneal administration of paclitaxel (PTX)-containing CM-NFBC efficiently suppressed tumor growth in a peritoneally disseminated cancer xenograft model. In this study, to confirm the applicability of NFBC in cancer therapy, a chemotherapeutic agent, doxorubicin (DXR), embedded into CM-NFBC, was examined for its efficiency to treat a peritoneally disseminated gastric cancer via intraperitoneal administration. DXR was efficiently embedded into CM-NFBC (DXR/CM-NFBC). In an in vitro release experiment, 79.5% of DXR was released linearly into the peritoneal wash fluid over a period of 24 h. In the peritoneally disseminated gastric cancer xenograft model, intraperitoneal administration of DXR/CM-NFBC induced superior tumor growth inhibition (TGI = 85.5%) by day 35 post-tumor inoculation, compared to free DXR (TGI = 62.4%). In addition, compared with free DXR, the severe side effects that cause body weight loss were lessened via treatment with DXR/CM-NFBC. These results support the feasibility of CM-NFBC as a drug-delivery vehicle for various anticancer agents. This approach may lead to improved therapeutic outcomes for the treatment of intraperitoneally disseminated cancers.

Download Full-text

An Orally Bioavailable, Indole-3-glyoxylamide Based Series of Tubulin Polymerization Inhibitors Showing Tumor Growth Inhibition in a Mouse Xenograft Model of Head and Neck Cancer

Journal of Medicinal Chemistry ◽

10.1021/acs.jmedchem.5b01312 ◽

2015 ◽

Vol 58 (23) ◽

pp. 9309-9333 ◽

Cited By ~ 35

Author(s):

Helen E. Colley ◽

Munitta Muthana ◽

Sarah J. Danson ◽

Lucinda V. Jackson ◽

Matthew L. Brett ◽

...

Keyword(s):

Head And Neck Cancer ◽

Tumor Growth ◽

Growth Inhibition ◽

Xenograft Model ◽

Tumor Growth Inhibition ◽

Tubulin Polymerization ◽

Mouse Xenograft ◽

Mouse Xenograft Model ◽

Orally Bioavailable ◽

Tubulin Polymerization Inhibitors

Download Full-text

Overexpression of Indoleamine 2,3-Dioxygenase in Human Endometrial Carcinoma Cells Induces Rapid Tumor Growth in a Mouse Xenograft Model

Clinical Cancer Research ◽

10.1158/1078-0432.ccr-08-0991 ◽

2008 ◽

Vol 14 (22) ◽

pp. 7251-7259 ◽

Cited By ~ 35

Author(s):

Norio Yoshida ◽

Kazuhiko Ino ◽

Yoshiyuki Ishida ◽

Hiroaki Kajiyama ◽

Eiko Yamamoto ◽

...

Keyword(s):

Tumor Growth ◽

Endometrial Carcinoma ◽

Xenograft Model ◽

Carcinoma Cells ◽

Mouse Xenograft ◽

Indoleamine 2,3 Dioxygenase ◽

Mouse Xenograft Model

Download Full-text

Prostate Tumor Growth and Recurrence Can Be Modulated by the ω-6:ω-3 Ratio in Diet: Athymic Mouse Xenograft Model Simulating Radical Prostatectomy

Neoplasia ◽

10.1593/neo.05637 ◽

2006 ◽

Vol 8 (2) ◽

pp. 112-124 ◽

Cited By ~ 67

Author(s):

Uddhav P. Kelavkar ◽

Justin Hutzley ◽

Rajiv Dhir ◽

Paul Kim ◽

Kenneth G.D. Allen ◽

...

Keyword(s):

Radical Prostatectomy ◽

Tumor Growth ◽

Xenograft Model ◽

Prostate Tumor ◽

Athymic Mouse ◽

Mouse Xenograft ◽

Mouse Xenograft Model

Download Full-text

Cyclopamine Reduces the Growth of Endometrial Carcinoma and Hedgehog Pathway Proteins in a Xenograft Mouse Model

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2500 ◽

2021 ◽

Vol 11 (3) ◽

pp. 420-425

Author(s):

Zhengxing He ◽

Jing Zeng ◽

Yalan Xu

Keyword(s):

Tumor Growth ◽

Receptor Antagonist ◽

Endometrial Carcinoma ◽

Mrna Expression ◽

Xenograft Model ◽

Female Reproductive Tract ◽

Tumor Growth Rate ◽

Inhibition Rate ◽

Transplanted Tumors ◽

Mouse Xenograft Model

Endometrial carcinoma is a frequently occurring malignancy of the female reproductive tract. Previous investigations have implicated the Hedgehog signaling pathway, including the smoothened (Smo) receptor, in tumor progression. Here, we established a nude mouse xenograft model of endometrial cancer to investigate the effect of the Smo receptor antagonist cyclopamine on the growth of endometrial carcinoma. Mice were randomly sorted into two groups receiving 0.2 mL/mouse every other day of either cyclopamine (20 mg/mL) or solvent (control). The growth of the mice was observed for 18 days. Then, mice were euthanized, tumors were removed and weighed, and tumor volume inhibition rate (VIR) and weight inhibition rate (WIR) were calculated. Smo, Gli1, and Gli2 mRNA expression was measured in tumor tissues by RT-PCR, and Vascular endothelial growth factor (VEGF) and CD31 were detected by immunohistochemistry. We found that cyclopamine treatment reduced the tumor growth rate and significantly reduced the volumes and weights of the transplanted tumors compared with those of the controls. Furthermore, the transplanted tumors of cyclopamine-treated mice possessed lower expression levels of Smo, Gli1, and Gli2, and displayed a lower positive expression of VEGF and CD31. Thus, the Smo receptor antagonist cyclopamine inhibited transplanted tumor growth in nude mice with endometrial carcinoma, which is likely connected with cyclopamine downregulation of Smo, Gli1, and Gli2 mRNA expression that promote angiogenesis.

Download Full-text

An Anti-PSMA Immunotoxin Reduces Mcl-1 and Bcl2A1 and Specifically Induces in Combination with the BAD-Like BH3 Mimetic ABT-737 Apoptosis in Prostate Cancer Cells

Cancers ◽

10.3390/cancers12061648 ◽

2020 ◽

Vol 12 (6) ◽

pp. 1648

Author(s):

Anie P. Masilamani ◽

Viviane Dettmer-Monaco ◽

Gianni Monaco ◽

Toni Cathomen ◽

Irina Kuckuck ◽

...

Keyword(s):

Prostate Cancer ◽

Combination Therapy ◽

Tumor Growth ◽

Cancer Cells ◽

Xenograft Model ◽

Target Cells ◽

Prostate Cancer Cells ◽

Mouse Xenograft Model ◽

Synergistic Cytotoxicity ◽

3D Spheroids

Background: Upregulation of anti-apoptotic Bcl-2 proteins in advanced prostate cancer leads to therapeutic resistance by prevention of cell death. New therapeutic approaches aim to target the Bcl-2 proteins for the restoration of apoptosis. Methods: The immunotoxin hD7-1(VL-VH)-PE40 specifically binds to the prostate specific membrane antigen (PSMA) on prostate cancer cells and inhibits protein biosynthesis. It was tested with respect to its effects on the expression of anti-apoptotic Bcl-2 proteins. Combination with the BAD-like mimetic ABT-737 was examined on prostate cancer cells and 3D spheroids and in view of tumor growth and survival in the prostate cancer SCID mouse xenograft model. Results: The immunotoxin led to a specific inhibition of Mcl-1 and Bcl2A1 expression in PSMA expressing target cells. Its combination with ABT-737, which inhibits Bcl-2, Bcl-xl, and Bcl-w, led to an induction of the intrinsic apoptotic pathway and to a synergistic cytotoxicity in prostate cancer cells and 3D spheroids. Furthermore, combination therapy led to a significantly prolonged survival of mice bearing prostate cancer xenografts based on an inhibition of tumor growth. Conclusion: The combination therapy of anti-PSMA immunotoxin plus ABT-737 represents the first tumor-specific therapeutic approach on the level of Bcl-2 proteins for the induction of apoptosis in prostate cancer.

Download Full-text

MNAT1 promotes proliferation and the chemo-resistance of osteosarcoma cell to cisplatin through regulating PI3K/Akt/mTOR pathway

BMC Cancer ◽

10.1186/s12885-020-07687-3 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Chensheng Qiu ◽

Weiliang Su ◽

Nana Shen ◽

Xiaoying Qi ◽

Xiaolin Wu ◽

...

Keyword(s):

Tumor Growth ◽

Xenograft Model ◽

Mtor Pathway ◽

Regulation Mechanism ◽

Osteosarcoma Cell ◽

Migration Ability ◽

Mouse Xenograft Model

Abstract Background MNAT1 (menage a trois 1, MAT1), a cyclin-dependent kinase-activating kinase (CAK) complex, highly expressed in diverse cancers and was involved in cancer molecular pathogenesis. However, its deliverance profile and biological function in osteosarcoma (OS) remain unclear. Methods The expression of MNAT1 in OS was detected by western blot (WB) and immunohistochemistry (IHC). The potential relationship between MNAT1 molecular level expression and OS clinical expectations were analyzed according to tissues microarray (TMA). Proliferation potential of OS cells was evaluated in vitro based on CCK8 and OS cells colony formation assays, while OS cells transwell and in situ tissue source wound healing assays were employed to analyze the OS cells invasion and migration ability in vitro. A nude mouse xenograft model was used to detect tumor growth in vivo. In addition, ordinary bioinformatics analysis and experimental correlation verification were performed to investigate the underlying regulation mechanism of OS by MNAT1. Results In this research, we found and confirmed that MNAT1 was markedly over-expressed in OS tissue derived in situ, also, highly MNAT1 expression was closely associated with bad clinical expectations. Functional studies had shown that MNAT1 silencing could weaken the invasion, migration and proliferation of OS cells in vitro, and inhibit OS tumor growth in vivo. Mechanism study indicated that MNAT1 contributed to the progression of OS via the PI3K/Akt/mTOR pathway. We further verified that the MNAT1 was required in the regulation of OS chemo-sensitivity to cisplatin (DDP). Conclusions Taken together, the data of the present study demonstrate a novel molecular mechanism of MNAT1 involved in the formation of DDP resistance of OS cells.

Download Full-text

VEGF/Flk1 Mechanism is Involved in Roxarsone Promotion of Rat Endothelial Cell Growth and B16F10 Xenograft Tumor Angiogenesis

Scientific Reports ◽

10.1038/s41598-019-53870-3 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Shihao Chen ◽

Jinge Xu ◽

Qianhan Wei ◽

Zeting Zhao ◽

Xin Chen ◽

...

Keyword(s):

Cell Growth ◽

Cell Viability ◽

Vascular Endothelial Cells ◽

Growth Promotion ◽

Xenograft Model ◽

Feed Additive ◽

Vascular Endothelial ◽

Mouse Xenograft Model ◽

Significant Difference

AbstractThe potential angiogenic effect of roxarsone, a feed additive widely used to promote animal growth worldwide, was demonstrated recently. We explored the mechanism of vascular endothelial growth factor (VEGF) and its receptor (VEGFR) in roxarsone promotion of rat vascular endothelial cells (ECs) and B16F10 mouse xenografts. ECs were treated with 0.1–50 μM roxarsone or with roxarsone plus 10 ng/mL VEGF, VEGFR1 (Flt1), or VEGFR2 (Flk1) antibodies for 12–48 h to examine their role in cell growth promotion. Small interfering RNA (siRNA) targeting Vegf, Flt1, and Flk1 were transfected in the ECs, and we measured the expression level, cell proliferation, migration, and tube formation ability. The siRNA targeting Vegf or Flk1 were injected intratumorally in the B16F10 xenografts of mice that received 25 mg/kg roxarsone orally. Cell viability and VEGF expression following roxarsone treatment were significantly higher than that of the control (P < 0.05), peaking following treatment with 1.0 μM roxarsone. Compared to roxarsone alone, the VEGF antibody decreased cell promotion by roxarsone (P < 0.05), and the Flk1 antibody greatly reduced cell viability compared to the Flt1 antibody (P < 0.01). Roxarsone and Flk1 antibody co-treatment increased supernatant VEGF significantly, while cellular VEGF was obviously decreased (P < 0.01), whereas there was no significant difference following Flt1 antibody blockade. The siRNA against Vegf or Flk1 significantly attenuated the roxarsone promotion effects on EC proliferation, migration, and tube-like formation (P < 0.01), whereas the siRNA against Flt1 effected no obvious differences. Furthermore, the RNA interference significantly weakened the roxarsone-induced increase in xenograft weight and volume, and VEGF and Flk1 expression. Roxarsone promotion of rat EC growth, migration, and tube-like formation in vitro and of B16F10 mouse xenograft model tumor growth and angiogenesis involves a VEGF/Flk1 mechanism.

Download Full-text