Journal: British Journal of Cancer

Article Title: Insulin-like growth factor binding protein-6 inhibits prostate cancer cell proliferation: implication for anticancer effect of diethylstilbestrol in hormone refractory prostate cancer

doi: 10.1038/sj.bjc.6602520

Figure Lengend Snippet: Inhibition of the proliferation of human prostate cancer cell lines LNCaP and PC-3 by diethylstilbestrol (DES). Cells were incubated for 24 h, and thereafter CM was aspirated away and the cells were incubated with CM containing various concentrations of DES. After 72 h, the number of viable cells was measured by MTT assay. Optical densities (OD) of cell lysates were measured at a wavelength of 540 nm. The values are expressed as means+s.d. ( n =3), and the P -values were <0.05 ( * ) and <0.01 (†).

Article Snippet: The human prostate cancer cell lines LNCaP and PC-3 were purchased from Dainippon Pharmaceutical (Tokyo, Japan) and cultured in RPMI (Sigma, St Louis, MO, USA) supplemented with 10% fetal calf serum (FCS) (Moregate, Bulimba, Australia).

Techniques: Inhibition, Incubation, MTT Assay

Journal: Toxins

Article Title: Curcumin―The Paradigm of a Multi-Target Natural Compound with Applications in Cancer Prevention and Treatment

doi: 10.3390/toxins2010128

Figure Lengend Snippet: Evaluation by gene expression profiling of the molecular targets of curcumin in cancer cells.

Article Snippet: Affymetrix, Superarray , human LNCap androgen-responsive prostate adenocarcinoma cell line, human C42B androgen non-responsive prostate adenocarcinoma cell line (derived from LNCap cell line) , Oxidative stress response was identified as the major pathway involved in curcumin induced biological responses in prostate cancer cells. Additionally curcumin suppresses androgen receptor in androgen responsive and refractory cells. , [ ] .

Techniques: Gene Expression, Expressing, Activity Assay, Knock-Out, Ubiquitin Proteomics, Quantitative Proteomics, Control, Derivative Assay

Journal: Therapeutic Advances in Medical Oncology

Article Title: Cotargeting Plk1 and androgen receptor enhances the therapeutic sensitivity of paclitaxel-resistant prostate cancer

doi: 10.1177/1758835919846375

Figure Lengend Snippet: Plk1 mRNA and protein with the active form were increased in paclitaxel-resistant cancer cells. (a–c) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to evaluate the mRNA levels of MDR1 (a), MRP1 (b), and Plk1 (c) in parental and paclitaxel-resistant LNCaP (LNCaP TXR ) or NCI-H460 (H460 TXR ) cells. Three independent experiments were performed. The relative expression of mRNA was plotted. ** p < 0.01, *** p < 0.001. (d) Cell lysates from parental and paclitaxel-resistant LNCaP (LNCaP TXR ) or NCI-H460 (H460 TXR ) cells were prepared to determine the levels of Plk1, Plk1 phosphorylated at T210, and actin proteins using specific antibodies. (e) The relative band intensity values of p-Plk1 (left panel) and Plk1 (right panel) were quantified with LI-COR Odyssey software and plotted.

Article Snippet: Human prostate cancer LNCaP (clone FGC, #21740), DU145 (#30081), and lung cancer NCI-H460 cells (#30177) were purchased from the Korean Cell Line Bank (KCLB; Seoul, Korea) and were authenticated before being frozen by KCLB using short tandem repeats (STR).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Software

Journal: Therapeutic Advances in Medical Oncology

Article Title: Cotargeting Plk1 and androgen receptor enhances the therapeutic sensitivity of paclitaxel-resistant prostate cancer

doi: 10.1177/1758835919846375

Figure Lengend Snippet: High levels of androgen receptor (AR) and prostate-specific antigen (PSA) in paclitaxel-resistant LNCaP TXR cells were markedly reduced by treatment with genistein. (a) Cells were treated with BI 2536 (left panel), volasertib (middle panel), or genistein (right panel) in a concentration-dependent manner in LNCaP, paclitaxel-resistant LNCaP (LNCaP TXR ), NCI-H460, and paclitaxel-resistant NCI-H460 (H460 TXR ) cells for 48 h. The numbers of viable cells were then measured by a cell viability assay. The bar graph presents the mean values of half maximal growth inhibitory concentration (GI 50 , nM for BI2536 and volasertib, μM for genistein). Three independent experiments were performed. * p < 0.05, ** p < 0.01, *** p < 0.001. (b) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to evaluate AR mRNA levels in LNCaP, LNCaP TXR , NCI-H460, and H460 TXR cells. Relative AR mRNA levels were evaluated in LNCaP TXR cells compared with that in LNCaP cells (left panel), and that in NCI-H460 TXR cells compared with in NCI-H460 cells (middle panel). Both AR and aldolase A mRNAs were detected in agarose gel after qRT-PCR (right panel). Three independent experiments were performed. (c) LNCaP and LNCaP TXR cells were grown for 48 h in the presence of 1, 10, 25, or 50 μM bicalutamide. The numbers of viable cells were then measured by cell viability assay. The bar graph presents the mean values of half maximal growth inhibitory concentration (GI 50 , μM) of bicalutamide in LNCaP and LNCaP TXR cells. (d-e) LNCaP and LNCaP TXR cells were grown for 48 h in the presence of 10 nM BI 2536, 15 nM volasertib, 30 µM genistein, or 14 µM bicalutamide. qRT-PCR was performed to evaluate mRNA levels of AR (d), and PSA (e) in LNCaP and LNCaP TXR cells. Three independent experiments were performed. Values are presented as the means ± standard deviations. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human prostate cancer LNCaP (clone FGC, #21740), DU145 (#30081), and lung cancer NCI-H460 cells (#30177) were purchased from the Korean Cell Line Bank (KCLB; Seoul, Korea) and were authenticated before being frozen by KCLB using short tandem repeats (STR).

Techniques: Concentration Assay, Viability Assay, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Agarose Gel Electrophoresis

Journal: Therapeutic Advances in Medical Oncology

Article Title: Cotargeting Plk1 and androgen receptor enhances the therapeutic sensitivity of paclitaxel-resistant prostate cancer

doi: 10.1177/1758835919846375

Figure Lengend Snippet: Genistein and bicalutamide are effective in paclitaxel-resistant DU145 TXR cells expressing high mRNA levels of androgen receptor (AR) and prostate-specific antigen (PSA). Paclitaxel-resistant DU145 TXR cells were developed as described in Materials and Methods. Genistein was treated in a concentration-dependent manner in DU145 and DU145 TXR cells for 48 h. The numbers of viable cells were then measured by a cell viability assay. The bar graph presents the mean values of half maximal growth inhibitory concentration (GI 50 , μM for genistein). (b) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to evaluate AR mRNA levels in DU145 and DU145 TXR cells. The mRNA levels of AR and aldolase A were detected in agarose gel after qRT-PCR (right panel) and those of LNCaP and LNCaP TXR cells were used as positive controls. (c) qRT-PCR was performed to evaluate mRNA levels of MDR1, MRP1, and Plk1 in DU145 and DU145 TXR cells. Three independent experiments were performed. (d) DU145 and DU145 TXR cells were grown for 48 h in the presence of BI 2536, volasertib, genistein, or bicalutamide. The numbers of viable cells were then measured by cell viability assay. The bar graph presents the mean values of half maximal growth inhibitory concentration (GI 50 , μM) of each compound in DU145 and DU145 TXR cells. (e) DU145 and DU145 TXR cells were grown for 48 h in the presence of 10 nM BI 2536, 15 nM volasertib, 30 µM genistein, or 14 µM bicalutamide. DU145 and DU145 TXR cells were grown for 48 h in the presence of 10 nM BI 2536, 15 nM volasertib, 30 µM genistein, or 14 µM bicalutamide. qRT-PCR was performed to evaluate mRNA levels of AR and PSA in DU145 and DU145 TXR cells. Three independent experiments were performed. Values are presented as the means ± standard deviations. ** p < 0.01, *** p < 0.001.

Article Snippet: Human prostate cancer LNCaP (clone FGC, #21740), DU145 (#30081), and lung cancer NCI-H460 cells (#30177) were purchased from the Korean Cell Line Bank (KCLB; Seoul, Korea) and were authenticated before being frozen by KCLB using short tandem repeats (STR).

Techniques: Expressing, Concentration Assay, Viability Assay, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Agarose Gel Electrophoresis

Journal: Therapeutic Advances in Medical Oncology

Article Title: Cotargeting Plk1 and androgen receptor enhances the therapeutic sensitivity of paclitaxel-resistant prostate cancer

doi: 10.1177/1758835919846375

Figure Lengend Snippet: Paclitaxel-resistant prostate cancer cells were sensitive to the treatment of genistein by reducing the activity of Plk1. (a) FACS analyses were performed on LNCaP and LNCaP TXR cells after treatment with 25 μM of genistein for 48 h. (b) The percentages of cells in the subG1 phase were measured by flow cytometry. * p < 0.05, ** p < 0.01, *** p < 0.001. (c) FACS analyses were performed on DU145 and DU145 TXR cells after treatment with 25 μM of genistein for 48 h. (d) The percentages of cells in the subG1 phase were measured by flow cytometry. * p < 0.05, ** p < 0.01, *** p < 0.001. (e) LNCaP and LNCaP TXR cells were grown for 48 h in the presence of 25 μM genistein. Cells were fixed with 4% paraformaldehyde and stained for active caspase-3 (green) and DAPI (blue). The percentage of cells that stained positive for active caspase-3 was determined. At least 1000 cells were counted, and three independent experiments were performed. * p < 0.05, ** p < 0.01, *** p < 0.001. (f) LNCaP and LNCaP TXR cells were grown for 48 h in the presence of 25 μM genistein. Lysates were analyzed by immunoblotting with anti-TCTP, anti-phospho-TCTP, anti-Plk1, anti-phospho-Plk1, and anti-actin antibodies (left panel). Ctrl, control. FACS, fluorescence-activated cell sorting; GS, genistein. The relative band intensity values of p-TCTP, TCTP, p-Plk1, and Plk1 were quantified with LI-COR Odyssey software and plotted (right panel).

Article Snippet: Human prostate cancer LNCaP (clone FGC, #21740), DU145 (#30081), and lung cancer NCI-H460 cells (#30177) were purchased from the Korean Cell Line Bank (KCLB; Seoul, Korea) and were authenticated before being frozen by KCLB using short tandem repeats (STR).

Techniques: Activity Assay, Flow Cytometry, Staining, Western Blot, Control, Fluorescence, FACS, Software