Journal: BioMed Research International

Article Title: Curcumin Targets Both Apoptosis and Necroptosis in Acidity-Tolerant Prostate Carcinoma Cells

doi: 10.1155/2021/8859181

Figure Lengend Snippet: Effects of docetaxel and curcumin on cell viability, ROS level, and mitochondrial membrane potential. PC-3 and PC-3AcT cells were treated with the indicated concentrations of DTX or CCM in DMEM containing 3.8 μ M lactic acid for 48 h. Two human prostate epithelial cell lines (RWPE-1 and HPrEC) were incubated in lactic acid-free DMEM. (a) Cell viability was measured by MTT assay. (b) Cellular ROS levels were measured by staining cells with DCF-DA (10 μ M). (c) ΔΨ m was measured by staining cells with Rhodamine 123 (30 nM). ∗ P < 0.05 vs. respective PC-3 cells. DTX: docetaxel; CCM: curcumin; ΔΨ m: mitochondrial membrane potential.

Article Snippet: The human PC cell line PC-3 and two human prostate epithelial cell lines, RWPE-1 and HPrEC, were obtained from the American Type Culture Collection (ATCC).

Techniques: Membrane, Incubation, MTT Assay, Staining

Journal: BioMed Research International

Article Title: Curcumin Targets Both Apoptosis and Necroptosis in Acidity-Tolerant Prostate Carcinoma Cells

doi: 10.1155/2021/8859181

Figure Lengend Snippet: Apoptosis- and necroptosis-inducing effects of curcumin. PC-3 and PC-3AcT cells were treated with the indicated concentrations of CCM for 48 h. Two human prostate epithelial cell lines (RWPE-1 and HPrEC) were incubated in lactic acid-free DMEM. (a) The levels of apoptosis- and necroptosis-mediated proteins were assessed by Western blotting. (b) Nuclear morphology was assessed by staining with DAPI (scale bar = 5 μ m). (c) Cells were pretreated with necrostatin-1 (25 μ M) and Q-VD-Oph-1 (10 μ M) 2 h prior to treatment with CCM (40 μ M) for 48 h in DMEM containing lactic acid (3.8 μ M). Cell viability was measured by MTT assay. ∗ P < 0.05 vs. respective control cells. # P < 0.05 vs. respective PC-3 cells. CCM: curcumin.

Article Snippet: The human PC cell line PC-3 and two human prostate epithelial cell lines, RWPE-1 and HPrEC, were obtained from the American Type Culture Collection (ATCC).

Techniques: Incubation, Western Blot, Staining, MTT Assay, Control

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Western blotting to detect the expression levels of LMNA and LMNB1 in cells. (A) The protein signal bands for LMNA, LMNB1, and GAPDH. (B, C) The relative expression levels of LMNB1 and LMNA (normalized to GAPDH expression) in different cell lines. The mean ± SEM of two independent experiments are shown (P = 0.0005 and P < 0.0001 for LMNB1 and LMNA, respectively; ANOVA). The results show that LMNB1-overexpressing EP156T (LMN-EP156T) cells express significantly higher amounts of LMNB1 than the parental and Mock EP156T cells. All three prostate cancer cell lines express high levels of LMNB1. All cells express LMNA, with PC-3 and DU145 expressing higher amounts of LMNA than the other cells.

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Western Blot, Expressing

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Immunohistochemical (IHC) staining of LMNB1 expression. Staining results were shown for (A) parental EP156T, (B) Mock EP156T, (C) LMNB1-overexpressing EP156T (LMN-EP156T), and (D) PC-3 cells (positive control). Both LMN-EP156T and PC-3 cells expressed strong nuclear staining of LMNB1, which was higher than either parental EP156T or Mock EP156T cells.

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Staining, Positive Control

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: MTS assay to measure the rate of cell growth. A total of 6.5 × 103 cells in 0.1 ml of complete medium were seeded onto each well of 96 well-plates in triplicate for each cell line. Five repeated plates were prepared for measurements at 5 time points. At each time point, one plate was used for this assay. MTS reagent (20 μl) was added to each well, and the plates were incubated at 37°C for 1 h. The absorbance at 490 nm is shown as the mean ± SEM of three independent experiments. LMNB1-overexpressing EP156T (LMN-EP156T) cells show significantly faster growth compared to the other two cell lines (P < 0.01; ANOVA).

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: MTS Assay, Incubation

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Matrigel-coated transwell assay (37°C for 48 h, 40 × magnification). A total of 5 × 104 cells of each cell line in 0.1 ml medium + 0.1% FBS were seeded onto an insert well (pore size, 8 µm) for a 24-well plate pre-coated with Matrigel. The insert wells were placed on receiver wells with 0.65 ml of medium + 10% FBS per well. After incubation at 37°C for 48 h, the cells on the apical side of the insert wells were removed with swabs. Only cells that had passed the membrane could be visualized after staining in 20% methanol with 0.25% crystal violet for 10 min. The results are shown for (A) EP156T, (B) Mock EP156T, and (C) LMN-EP156T cells. (D) The number of cells that penetrated the wells are shown as the mean ± SEM of three independent experiments. Statistical significance was determined by ANOVA (P < 0.0001). Significantly more LMNB1-overexpressing EP156T (LMN-EP156T) cells passed through the 8-µm pores than parental and Mock EP156T cells.

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Transwell Assay, Pore Size, Incubation, Membrane, Staining

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Colony-formation assay. Results are shown for (A) EP156T, (B) Mock EP156T, (C) LMNB1-overexpressing EP156T (LMN-EP156T), and (D) PC-3 cells. A total of 5 × 103 cells of each cell line were mixed in 0.3% soft agar with complete growth medium, and plated onto a culture dish (diameter, 3.5 cm). Numerous colonies can be seen in the plates inoculated with PC-3 after 2 weeks of incubation (magnification, 100 ×) at 37°C. However, no colonies have formed in the plates inoculated with the other cell lines. White arrows indicate PC-3 cell colonies.

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Colony Assay, Incubation

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Xenograft mouse model showing evident subcutaneous tumors in the right chest 7 weeks after the injection of PC-3 cells. (A-D) Four of the five mice injected with PC-3 cells formed subcutaneous tumors (red arrows). The tumor sizes range between 13 mm and 19 mm in diameter. No tumors were seen in mice injected with EP156T, Mock EP156T, or LMNB1-overexpressing EP156T cells (photos not shown).

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Injection

Journal: American Journal of Cancer Research

Article Title: LMNB1, a potential marker for early prostate cancer progression

doi:

Figure Lengend Snippet: Hematoxylin and eosin-stained tissue sections from xenografted mice. (A) An enlarged and metastasized lymph node from the PC-3-injected mouse. (B-E) Lung sections from mice injected with PC-3, EP156T, Mock EP156T, and LMNB1-overexpressing EP156T cells, respectively. Each row shows the same section with increasing magnification. Red arrows indicate PC-3 metastatic cell nests with clear nucleoli in either the lymph node (A) or lung (B). There are no subcutaneous growths or metastatic lesions in mice injected with the parental, Mock, or LMNB1-overexpressing EP156T cells.

Article Snippet: EP156T prostate epithelial cells and transfection with lentiviral vectors harboring LMNB1 A human TERT -immortalized EP156T prostatic epithelial cell line was purchased from the American Type Culture Collection (CRL3289TM, ATCC, Manassas, VA, USA) and grown in MCDB-153 medium supplemented with bovine pituitary extract (25 mg/500 ml medium), hEGF (5 ng/ml medium), 1% fetal bovine serum (FBS), and 0.5 μg/ml puromycin.

Techniques: Staining, Injection