human pc cell lines in vitro du145  (ATCC)

Journal: Cancer Cell International

Article Title: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1

doi: 10.1186/s12935-020-01481-8

Figure Lengend Snippet: MiR-15a negatively regulated the expression of PD-L1. a , b The expression levels of miR-15a ( a ) and PD-L1 ( b ) were examined by RT-qPCR. c The protein level of PD-L1 was examined by western blot. DU145 and PC-3 were transfected with miR-15a mimics, miR-15a inhibitor, or the corresponding control miRNA (mimics NC or inhibitor NC). d Bioinformatic analysis using Starbase ( http://starbase.sysu.edu.cn/index.php ) revealed potential binding sites between miR-15a and PD-L1. e The relative luciferase activities were measured and compared between cells co-transfected with miR-15a mimics or miR-15a inhibitor. DU145 or PC-3 cells were transfected with luciferase reporter gene driven by either the wild type of PD-L1 (WT-PD-L1) or the mutated PD-L1 (MUT-PD-L1). *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: The human PC cell lines DU145 and PC-3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Control, Binding Assay, Luciferase

Journal: Cancer Cell International

Article Title: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1

doi: 10.1186/s12935-020-01481-8

Figure Lengend Snippet: PD-L1 played a central role in antagonizing miR-15a-induced cytotoxicity of CD8 + T cells. a , b The expression levels of miR-15a ( a ) and PD-L1 ( b ) were examined by RT-qPCR. c The protein level of PD-L1 was examined by western blot. d The cytotoxicity of CD8 + T cells was measured using LDH Kit. e The proliferation of CD8 + T cells was measured by CFSE assay followed by flow cytometry. f The apoptosis of CD8 + T cells was examined by Annexin V/PI dual staining followed by flow cytometry. The effector CD8 + T cells were isolated from peripheral blood of healthy donors and co-cultured with indicated target PC cells. DU145 and PC-3 cells were not transfected or transfected with miR-15a mimics, miR-15a mimics + pcDNA3.1-NC, or miR-15a mimics + pcDNA3.1-PD-L1, respectively. *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: The human PC cell lines DU145 and PC-3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CFSE Assay, Flow Cytometry, Staining, Isolation, Cell Culture, Transfection

Journal: Cancer Cell International

Article Title: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1

doi: 10.1186/s12935-020-01481-8

Figure Lengend Snippet: MiR-15a regulated multiple malignant phenotypes of PC cells via directly targeting PD-L1. a – d Cell viability ( a ), apoptosis ( b ), migration ( c ), and invasion ( d ) of indicated PC cells were examined by MTT assay, Annexin V/PI staining followed by flow cytometry, wound healing assay, or Transwell assay, respectively. e The protein levels of Ras, p-ERK1/2, ERK1/2, Snail, E-cadherin, N-cadherin, MMP-9 from indicated PC cells were examined by western blot. DU145 and PC-3 cells were not transfected or transfected with miR-15a mimics, miR-15a mimics + pcDNA3.1-NC, or miR-15a mimics + pcDNA3.1-PD-L1, respectively. *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: The human PC cell lines DU145 and PC-3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Migration, MTT Assay, Staining, Flow Cytometry, Wound Healing Assay, Transwell Assay, Western Blot, Transfection

Journal: Cancer Cell International

Article Title: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1

doi: 10.1186/s12935-020-01481-8

Figure Lengend Snippet: KCNQ1OT1 negatively regulated the expression of miR-15a and inhibited the cytotoxicity of CD8 + T cells. a , b The expression levels of KCNQ1OT1 ( a ) and miR-15a ( b ) were examined by RT-qPCR. DU145 or PC-3 cells were transfected pcDNA3.1-NC, pcDNA3.1-KCNQ1OT1, shNC, or shKCNQ1OT1. Non-transfected cells were used as control. c Bioinformatic analysis using Starbase ( http://starbase.sysu.edu.cn/index.php ) revealed potential binding sites between KCNQ1OT1 and miR-15a. d The relative luciferase activities were measured and compared between cells co-transfected with miR-15a mimics or miR-15a inhibitor. DU145 or PC-3 cells were transfected with luciferase reporter gene driven by either the wild type of KCNQ1OT1 (WT-KCNQ1OT1) or the mutated KCNQ1OT1 (MUT-KCNQ1OT1). e The protein level of PD-L1 was measured in indicated cells using western blot. f – h The cytotoxicity ( f ), proliferation ( g ), and apoptosis ( h ) of CD8 + T cells upon co-cultured with indicated PC cells were measured as described in Fig. . DU145 and PC-3 cells were not transfected or transfected with shNC, shKCNQ1OT1, shKCNQ1OT1 + inhibitor NC, or shKCNQ1OT1 + miR-15a inhibitor, respectively. *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: The human PC cell lines DU145 and PC-3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Binding Assay, Luciferase, Western Blot, Cell Culture

Journal: Cancer Cell International

Article Title: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1

doi: 10.1186/s12935-020-01481-8

Figure Lengend Snippet: KCNQ1OT1 essentially maintained the malignant phenotypes of PC cells by targeting miR-15a. a – c The apoptosis ( a ), migration ( b ), and invasion ( c ) of indicated PC cells were examined by Annexin V/PI dual staining followed by flow cytometry, wound healing assay, and Transwell assay, respectively. d The protein levels of Ras, p-ERK1/2, ERK1/2, Snail, E-cadherin, N-cadherin and MMP-9 were examined by western blot. DU145 and PC-3 cells were not transfected or transfected with shNC, shKCNQ1OT1, shKCNQ1OT1 + inhibitor NC, or shKCNQ1OT1 + miR-15a inhibitor, respectively. *P < 0.05 and **P < 0.01

Article Snippet: The human PC cell lines DU145 and PC-3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA) and 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37 °C.

Techniques: Migration, Staining, Flow Cytometry, Wound Healing Assay, Transwell Assay, Western Blot, Transfection

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of suberoylanilide hydroxamic acid (SAHA) and Rosmarinic acid (RA) on cell viability in PC-3 and DU145 cell lines. After PC-3 and DU145, cell lines were seeded at 1 × 10 4 cells per well in 96-well plates and treated with media containing negative control (NC; DMSO), SAHA (1, 2.5, 5, 10, 25, 50 μM) and RA (25, 50, 100, 200, 250, 300 μM), cell viability of each cell line was evaluated. The data showed ( A ) the effect of SAHA and ( B ) the effect of RA on cell viability of both cell lines. The results are expressed as means ± standard deviation (SD). * p < 0.05: a significant difference versus control.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Negative Control, Standard Deviation, Control

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on formation of colonies and tumor spheroids in PC-3 and DU145 cell lines. ( A ) After both cell lines were seeded at 5 × 10 3 cells per well in 6-well plates and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), colonogenic assay was performed to measure the colony formation. ( B ) The number of colonies was quantified by using the Image J program. ( C ) Both cell lines in media containing NC (DMSO), SAHA (1 μM) and RA (200 μM) were seeded on petri dish covers at 3 × 10 3 cells in 25 μL. ( D ) The size of tumor spheroids was measured by using the Image J program. The results are expressed as means ± SD. * p < 0.05: a significant difference versus control.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Control

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on the apoptotic events in PC-3 and DU145 cell lines. ( A ) After both cell lines were seeded at 7 × 10 5 cells in cell culture dishes and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), Annexin V assay was conducted. ( B ) The apoptotic cells at each stage were analyzed and quantified by using the flow cytometry. Q1, Q2, Q3 and Q4 indicate necrosis, late apoptosis, live and early apoptosis, respectively.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Cell Culture, Annexin V Assay, Flow Cytometry

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on DNA fragmentation in PC-3 and DU145 cell lines. After both cell lines were seeded at 3 × 10 5 cells per well in 24-well plates and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed. ( A ) The data indicated that nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), and DNA fragmentation was stained with TUNEL. DAPI and TUNEL were merged to observe apoptotic nuclei. ( B ) The apoptotic cells presenting DNA fragmentation were quantified separately. The results are expressed as means ± SD. * p < 0.05: a significant difference versus control. MERGE means the combined picture of DAPI and TUNEL pictures.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: TUNEL Assay, Staining, Control

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on expression of histone deacetylase 2 (HDAC2) and p53 in PC-3 and DU145 cell lines. After both cell lines were seeded at 1 × 10 6 in cell culture dishes and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), the western blot analysis was performed. ( A ) The expressions of HDAC2 and p53 at the protein level were confirmed by western blot analysis. ( B ) The expression levels of HDAC2 and p53 were quantified and normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The results are expressed as means ± SD. * p < 0.05: a significant difference versus control.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Histone Deacetylase Assay, Cell Culture, Western Blot, Control

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on expression of cell cycle related genes in PC-3 and DU145 cell lines. After both cell lines were seeded at 1 × 10 6 in cell culture dishes and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), the western blot analysis was performed. ( A ) The expressions of p21, proliferating cell nuclear antigen (PCNA), cyclin D1 and cyclin E1 at the protein level were confirmed by western blot analysis. ( B ) The expression levels of each gene were quantified and normalized to GAPDH. The results are expressed as means ± SD. * p < 0.05: a significant difference versus control.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: Nutrients

Article Title: Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines

doi: 10.3390/nu10111784

Figure Lengend Snippet: Effects of SAHA and RA on expression of apoptosis related genes in PC-3 and DU145 cell lines. After both cell lines were seeded at 1 × 10 6 in cell culture dishes and treated with media containing NC (DMSO), SAHA (1 μM) and RA (200 μM), the western blot analysis was performed. ( A ) The expressions of Bax, Bcl-2, caspase-3 and poly [ADP-ribose] polymerase 1 (PARP-1) (cleaved) at the protein level were confirmed by western blot analysis. ( B ) The expression levels of each gene were quantified and normalized to GAPDH. The results are expressed as means ± SD. * p < 0.05: a significant difference versus control.

Article Snippet: The human PCa cell lines, PC-3 and DU145, were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Cell Culture, Western Blot, Control