human prostate cancer cell line lncap  (ATCC)

Journal: International Journal of Biological Sciences

Article Title: TRPV1 Downregulation Impairs Prostate Cancer Growth: Functional and Translational Insights from Cellular and In Vivo Models

doi: 10.7150/ijbs.125429

Figure Lengend Snippet: TRPV1 expression in prostate cell lines. (A) TRPV1 expression levels were determined by RT-qPCR. Data represent relative expression normalized to actin, used as the housekeeping gene. Results are presented as mean ± SD of three independent experiments. (B) TRPV1 protein expression was analyzed by Western blotting, with GAPDH as a loading control. The healthy prostate cell line PNT2 was used for comparison. A representative blot from three independent experiments is shown. (C) Fluorescence microscopy images showing TRPV1 labeling (green) in PNT2, LNCaP, LN-FLU, PC3, and DU-145 cells. Nuclei were stained with DAPI (blue). (D) Top row: lysosomes labeled with LysoTracker (red) and nuclei stained with DAPI (blue). Bottom row: PC3 cells transfected with CellLight ER-RFP BacMam to visualize the endoplasmic reticulum. A representative image from two independent experiments is shown. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****) indicate significant differences via one-way ANOVA.

Article Snippet: The human prostate cancer cell lines LNCaP (CRL-1740), PC3 (CRL-1435), and DU-145 (HTB-81) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Comparison, Fluorescence, Microscopy, Labeling, Staining, Transfection

Journal: International Journal of Biological Sciences

Article Title: TRPV1 Downregulation Impairs Prostate Cancer Growth: Functional and Translational Insights from Cellular and In Vivo Models

doi: 10.7150/ijbs.125429

Figure Lengend Snippet: Effect of TRPV1 receptor silencing on cell viability and proliferation. LNCaP, PC3, and DU-145 cells were infected with either an empty vector (EV) or a shTRPV1 vector via lentivirus-mediated transduction. (A) Cell viability of prostate cancer cells. Following infection, the cells were seeded at equal densities, and MTT assays were performed at 12, 24, and 48 hours post-seeding to evaluate the impact of TRPV1 silencing on cell proliferation. Data from non-silenced and silenced cells were normalized to 1 to facilitate the comparison. The mean ± SD of three independent experiments is shown. (B) Protein expression levels of proteins involved in proliferation, cell cycle regulation and mitosis. The levels of the proteins were determined by Western blotting, and β-actin served as a loading control. The densitometric analyses of the bands represent the mean ± SD of three different experiments. (C) Relative mRNA expression levels of the stemness markers Oct4, Nanog and ABCB1A. mRNA levels were quantified via RT-qPCR, normalized to actin (housekeeping gene) and presented as mean ± SD of six independent experiments. (D) Correlation analysis between TRPV1 and the stemness markers Oct4, Nanog and ABCB1A in TRPV1-silenced cells. Top: Scatter plots showing Pearson's correlation coefficients (r) for TRPV1 vs. Oct4, TRPV1 vs. Nanog and TRPV1 vs. ABCB1A. Bottom: Heatmap displaying Pearson's correlation coefficients for TRPV1, Oct4, Nanog and ABCB1A expression. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****) indicate significant differences according to two-way ANOVA and Sidak's multiple comparisons test.

Article Snippet: The human prostate cancer cell lines LNCaP (CRL-1740), PC3 (CRL-1435), and DU-145 (HTB-81) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Infection, Plasmid Preparation, Transduction, Comparison, Expressing, Western Blot, Control, Quantitative RT-PCR