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YAP is closely correlated with castration-resistant prostate cancer, and downregulation of YAP reduces proliferation and induces apoptosis of PC-3 cells.

Sheng X, Li WB, Wang DL, Chen KH, Cao JJ, Luo Z, He J, Li MC, Liu WJ, Yu C - Mol Med Rep (2015)

Bottom Line: The frequency of cells that were positive for YAP protein in PCa (78.13%) was significantly higher, compared with para‑PCa (26.67%; P=0.007) and benign prostatic hyperplasia (0%; P=0.002).The proliferative capacity of the transfected group was significantly lower, compared with the negative control group (P=0.022).Therefore, it was concluded that gene transcription and protein expression of YAP may be involved in the development of PCa, particularly CRPC, and may be a novel biomarker for investigation of the occurrence and progression of CRPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT
Yes-associated protein 65 (YAP65) has been implicated as an oncogene, and its expression is increased in human cancer. Previous studies have demonstrated that alterations in YAP activity may result in tumourigenesis of the prostate. With androgen deprivation therapies becoming progressively ineffective, often leading to life‑threatening androgen‑resistant prostate cancer (CRPC). The present study aimed to analyse the role of YAP in prostate cancer (PCa), particularly in CRPC. YAP protein was detected using immunohistochemistry and western blot analysis in different prostatic tissues. In addition, three specific RNA interference vectors targeting the human YAP gene were synthesised, and PC‑3 cells with a stable inhibition of YAP were obtained by transfection. MTT, flow cytometry, reverse transcription‑quantitative polymerase chain reaction and western blot assays were used to analyse the effects of YAP inhibition on the proliferation and apoptosis of PC‑3 cells. The frequency of cells that were positive for YAP protein in PCa (78.13%) was significantly higher, compared with para‑PCa (26.67%; P=0.007) and benign prostatic hyperplasia (0%; P=0.002). The frequency of cells, which were positive for the expression of YAP exhibited a positive correlation (P=0.008) with the Gleason score, the tumour‑node‑metastasis staging (P=0.033) and the level of prostate specific antigens (P=0.0032) in PCa. The proliferative capacity of the transfected group was significantly lower, compared with the negative control group (P=0.022). The cell‑cycle of the transfected group was arrested in the G1 stage, which was detected using flow cytometry, and there was a significant increase in the apoptosis of cells in the transfected group (P=0.002). The mRNA and protein levels of TEA domain family member 1 were inhibited in the transfected group (P=0.001 and P=0.00, respectively). Therefore, it was concluded that gene transcription and protein expression of YAP may be involved in the development of PCa, particularly CRPC, and may be a novel biomarker for investigation of the occurrence and progression of CRPC. However, the mechanism underlying the modulation of YAP in CRPC remains to be fully elucidated.

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Related in: MedlinePlus

Transfection assay using a fluorescent plasmid. The expression of green fluorescent protein with interference plasmid transfected PC-3 cells was assessed. (A) Imaging under an inverted microscope (magnification, ×200) following transfection for 48 h; (B) Cells following transfection for 48 h under an inverted fluorescence microscope (magnification, ×200); (C) Fluorescent cell screening 2 weeks following treatment with puromycin (magnification, ×200); (D) Fluorescent cell screening after 4 weeks (magnification, ×200), which were considered to be stably expressing green fluorescence.
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f3-mmr-12-04-4867: Transfection assay using a fluorescent plasmid. The expression of green fluorescent protein with interference plasmid transfected PC-3 cells was assessed. (A) Imaging under an inverted microscope (magnification, ×200) following transfection for 48 h; (B) Cells following transfection for 48 h under an inverted fluorescence microscope (magnification, ×200); (C) Fluorescent cell screening 2 weeks following treatment with puromycin (magnification, ×200); (D) Fluorescent cell screening after 4 weeks (magnification, ×200), which were considered to be stably expressing green fluorescence.

Mentions: GFP was expressed in the PC-3 cells 48 h after transfection with the pMagic7.1-Puro/GFP-RNAi-1, 2, and 3 plasmids, which suggested that each group was transfected successfully (Fig. 3A and B). After 2 weeks of selection with 1 mg/l puromycin, small clusters of green fluorescent cells were observed (Fig. 3C). Subsequently, three groups of stable YAP interference cell lines, as well as one non-targeted knockdown control cell line, were established from the PC-3 cells (Fig. 3D).


YAP is closely correlated with castration-resistant prostate cancer, and downregulation of YAP reduces proliferation and induces apoptosis of PC-3 cells.

Sheng X, Li WB, Wang DL, Chen KH, Cao JJ, Luo Z, He J, Li MC, Liu WJ, Yu C - Mol Med Rep (2015)

Transfection assay using a fluorescent plasmid. The expression of green fluorescent protein with interference plasmid transfected PC-3 cells was assessed. (A) Imaging under an inverted microscope (magnification, ×200) following transfection for 48 h; (B) Cells following transfection for 48 h under an inverted fluorescence microscope (magnification, ×200); (C) Fluorescent cell screening 2 weeks following treatment with puromycin (magnification, ×200); (D) Fluorescent cell screening after 4 weeks (magnification, ×200), which were considered to be stably expressing green fluorescence.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581809&req=5

f3-mmr-12-04-4867: Transfection assay using a fluorescent plasmid. The expression of green fluorescent protein with interference plasmid transfected PC-3 cells was assessed. (A) Imaging under an inverted microscope (magnification, ×200) following transfection for 48 h; (B) Cells following transfection for 48 h under an inverted fluorescence microscope (magnification, ×200); (C) Fluorescent cell screening 2 weeks following treatment with puromycin (magnification, ×200); (D) Fluorescent cell screening after 4 weeks (magnification, ×200), which were considered to be stably expressing green fluorescence.
Mentions: GFP was expressed in the PC-3 cells 48 h after transfection with the pMagic7.1-Puro/GFP-RNAi-1, 2, and 3 plasmids, which suggested that each group was transfected successfully (Fig. 3A and B). After 2 weeks of selection with 1 mg/l puromycin, small clusters of green fluorescent cells were observed (Fig. 3C). Subsequently, three groups of stable YAP interference cell lines, as well as one non-targeted knockdown control cell line, were established from the PC-3 cells (Fig. 3D).

Bottom Line: The frequency of cells that were positive for YAP protein in PCa (78.13%) was significantly higher, compared with para‑PCa (26.67%; P=0.007) and benign prostatic hyperplasia (0%; P=0.002).The proliferative capacity of the transfected group was significantly lower, compared with the negative control group (P=0.022).Therefore, it was concluded that gene transcription and protein expression of YAP may be involved in the development of PCa, particularly CRPC, and may be a novel biomarker for investigation of the occurrence and progression of CRPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT
Yes-associated protein 65 (YAP65) has been implicated as an oncogene, and its expression is increased in human cancer. Previous studies have demonstrated that alterations in YAP activity may result in tumourigenesis of the prostate. With androgen deprivation therapies becoming progressively ineffective, often leading to life‑threatening androgen‑resistant prostate cancer (CRPC). The present study aimed to analyse the role of YAP in prostate cancer (PCa), particularly in CRPC. YAP protein was detected using immunohistochemistry and western blot analysis in different prostatic tissues. In addition, three specific RNA interference vectors targeting the human YAP gene were synthesised, and PC‑3 cells with a stable inhibition of YAP were obtained by transfection. MTT, flow cytometry, reverse transcription‑quantitative polymerase chain reaction and western blot assays were used to analyse the effects of YAP inhibition on the proliferation and apoptosis of PC‑3 cells. The frequency of cells that were positive for YAP protein in PCa (78.13%) was significantly higher, compared with para‑PCa (26.67%; P=0.007) and benign prostatic hyperplasia (0%; P=0.002). The frequency of cells, which were positive for the expression of YAP exhibited a positive correlation (P=0.008) with the Gleason score, the tumour‑node‑metastasis staging (P=0.033) and the level of prostate specific antigens (P=0.0032) in PCa. The proliferative capacity of the transfected group was significantly lower, compared with the negative control group (P=0.022). The cell‑cycle of the transfected group was arrested in the G1 stage, which was detected using flow cytometry, and there was a significant increase in the apoptosis of cells in the transfected group (P=0.002). The mRNA and protein levels of TEA domain family member 1 were inhibited in the transfected group (P=0.001 and P=0.00, respectively). Therefore, it was concluded that gene transcription and protein expression of YAP may be involved in the development of PCa, particularly CRPC, and may be a novel biomarker for investigation of the occurrence and progression of CRPC. However, the mechanism underlying the modulation of YAP in CRPC remains to be fully elucidated.

Show MeSH
Related in: MedlinePlus