Establishment of An Orthotopic Prostate Cancer Xenograft Mouse Model Using Microscope-Guided Orthotopic Injection of LNCaP Cells Into The Dorsal Lobe of The Mouse Prostate

Background: Orthotopic LNCaP xenograft mouse models closely mimic the progression of androgen-dependent prostate cancer in humans; however, orthotopic injection of LNCaP cells into the mouse prostate remains a challenge.Methods: Under the guidance of a stereoscopic microscope, the anatomy of the individual prostate lobes in male Balb/c athymic nude mice was investigated, and LNCaP cells were inoculated into the mouse dorsal prostate (DP) to generate orthotopic tumors that mimicked the pathophysiological process of prostate cancer in humans. Real-time ultrasound imaging was used to monitor orthotopic prostate tumorigenesis, contrast-enhanced ultrasonography (CEUS) was used to characterize tumor angiogenesis, and macroscopic and microscopic characteristics of tumors were described.Results: The DP had a trigonal bipyramid-shape and were located at the base of the seminal vesicles. After orthotopic inoculation, gray scale ultrasound imaging showed progressive changes in tumor echotexture, shape and location, and tumors tended to protrude into the bladder. After 8 weeks, the tumor take rate was 65% (n= 13/20 mice). On CEUS, signal intensity increased rapidly, peaked, and decreased gradually. Observations of gross specimens showed orthotopic prostate tumors were well circumscribed, round, dark brown, and soft, with a smooth outer surface and a glossy appearance. Microscopically, tumor cells were arranged in acini encircled by fibrous septa with variably thickened walls, mimicking human adenocarcinoma.Conclusions: This study describes a successful approach to establishing an orthotopic LNCaP xenograft Balb/c athymic nude mouse model. The model requires a thorough understanding of mouse prostate anatomy and proper technique. The model represents a valuable tool for the in vivo study of the biological processes involved in angiogenesis in prostate cancer and preclinical evaluations of novel anti-angiogenic therapies.

In Vivo Phytotherapy in BALB/c Athymic Nude Mice: Hair Growth Promotion using Ficus religiosa L. and Morus alba L.

<em>Ficus religiosa</em> L. (FR) and <em>Morus alba</em> L. (ML) belonging to the family Moraceae have been tested as novel herbal agents for hair growth promotion and Hair Follicles (HFs) regeneration in BALB/c athymic nude mouse model. Current study tested different mixtures of 5% aqueous fractions: Test 1 (ML2+ML3+ML4+FR4), Test 2 (FR1+FR2+FR4), or Test 3 (ML2+ML3+ML4+FR1+FR2+FR4) from leaves of both plants including standard of care 2% minoxidil. Control mice were untreated. Animals were treated for 33 days by topical application on the back skin and changes in hair growth patterns were evaluated. Histology was performed to assess the HFs morphology, and modulation of hair cycle phases. Gene expression analysis was performed to understand potential mechanisms of action. All treatment groups had significantly higher anagen phase HFs compared with untreated control group based on histology analysis. Also, expressions of CD34, CD200R and Oct4 genes were upregulated in all treatment groups compared with untreated control. Present study demonstrated that a combinatorial therapy using either fractions of FR or FR and ML promoted hair growth and HFs regeneration through induction of anagen phase in conjunction with stem cells associated genes upregulation in BALB/c athymic nude mouse model of hair loss.

Histone Deacetylase Inhibitor SAHA Is a Promising Treatment of Cushing Disease

Context Remission failure following transsphenoidal surgery in Cushing disease (CD) from pituitary corticotroph tumors (CtTs) remains clinically challenging. Histone deacetylase inhibitors (HDACis) are antitumor drugs approved for clinical use, with the potential to affect adrenocorticotropin hormone (ACTH) hypersecretion by inhibiting pro-opiomelanocortin (POMC) transcription. Objective Testing the efficacy of suberoylanilide hydroxamic acid (SAHA) on human and murine ACTH-secreting tumor (AtT-20) cells. Design Cell viability, ACTH secretion (enzyme-linked immunosorbent assay), apoptosis, and gene expression profile were investigated on AtT-20 cells. In vivo efficacy was examined in an athymic nude mouse AtT-20 xenograft model. SAHA efficacy against human-derived corticotroph tumor (hCtT) (n = 8) was tested in vitro. Setting National Institutes of Health. Intervention SAHA (0.5 to 8 µM). Main Outcome Measures AtT-20 and hCtT cell survival, in vitro/invivo ACTH measurements. Results SAHA (1 µM) reduced AtT-20 viability to 75% at 24 hours, 43% at 48 hours (analysis of variance; P = 0.002). Apoptosis was confirmed with elevated BAX/Bcl2 ratio and FACS. Intriguingly, early (3-hour) significant decline (70%; P &lt; 0.0001) of secreted ACTH and diminished POMC transcription was observed with SAHA (1 µM). Microarray analysis revealed a direct association between liver X receptor alpha (LXRα) and POMC expression. Accordingly, SAHA reduced LXRα in AtT-20 cells but not in normal murine corticotrophs. Xenografted nude-mice tumor involution (126 ± 33/160 ± 35 vs 337 ± 49 mm3; P = 0.0005) was observed with 5-day intraperitoneal SAHA, with reversal of elevated ACTH (P &lt; 0.0001). SAHA did not affect serum ACTH in nontumor mice. Lastly, we confirmed that SAHA (1 µM/24 h) decreased hCtT survival (78.92%; P = 0.0007) and ACTH secretion (83.64%; P = 0.03). Conclusion Our findings demonstrate SAHA’s efficacy in reducing survival and ACTH secretion in AtT-20 and hCtT cells, providing a potential intervention for recurrent/unremitting CD.