Proteomic analysis of androgen-independent growth in low and high passage human LNCaP prostatic adenocarcinoma cells

Youm, Yun‐Hee; Kim, Seyoon; Bahk, Young Yil; Yoo, Tag Keun

doi:10.5483/bmbrep.2008.41.10.722

Cited by 7 publications

(5 citation statements)

References 26 publications

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“…The findings are intriguing and suggest that progressive sub-culturing of cell lines alters gene expression, a finding reported by others [38–39]. For example, continued passage of the LNCaP prostatic adenocarcinoma cell line has been shown to mimic the transformation of prostatic cancer from an androgen-dependent to an androgen-independent phenotype [40]. Thus, it is possible that the varying reports of Thy 1.2 expression in the RGC-5 cells reflect alterations due to continued sub-culturing of the cell line.…”

Section: Discussionmentioning

confidence: 87%

Sensitivity of Staurosporine-Induced Differentiated RGC-5 Cells to Homocysteine

Ganapathy

Dun

et al. 2009

Current Eye Research

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PURPOSE Homocysteine is implicated in ganglion cell death associated with glaucoma. To understand mechanisms of homocysteine-induced cell death, we analyzed the sensitivity of the RGC-5 cell line, differentiated using staurosporine, to physiologically-relevant levels of the excitotoxic amino acid homocysteine. METHODS RGC-5 cells were differentiated 24 h using 316 nM staurosporine and tested for expression of Thy 1.2 via immunodetection, RT-PCR and immunoblotting. The sensitivity of staurosporine-differentiated RGC-5 cells to physiological levels of homocysteine (50, 100, 250 µM) and to high levels of homocysteine (1 mM), glutamate (1 mM) and oxidative stress (25 µM:10 mU/ml xanthine:xanthine oxidase) was assessed by TUNEL assay and by immunodetection of cleaved caspase-3. The sensitivity of undifferentiated RGC-5 cells to high (1, 5, and 10 mM) homocysteine was also examined. RESULTS Undifferentiated RGC-5 cells express Thy 1.2 mRNA and protein. Staurosporine-differentiated RGC-5 cells extend neurite processes and express Thy 1.2 after 24 h differentiation; they express NF-L after 1 and 3 days differentiation. Treatment of staurosporine-differentiated RGC-5 cells with 50, 100 or 250µM homocysteine did not alter neurite processes nor induce cell death (detected by TUNEL and active caspase-3) to a level greater than that observed in non-homocysteine-treated, staurosporine-differentiated cells. The 1 mM dosage of homocysteine in staurosporine-differentiated RGC-5 cells also did not induce cell death above control levels, although 18 h treatment of non-differentiated RGC-5 cells with 5 mM homocysteine decreased survival by 50%. CONCLUSIONS RGC-5 cells differentiated for 24 h with 316 nM staurosporine project robust neurite processes and are positive for ganglion cell markers consistent with a more neuronal phenotype than non- staurosporine-differentiated RGC-5 cells. However, concentrations of homocysteine known to induce ganglion cell death in vivo and in primary ganglion cells are not sufficient to induce death of RGC-5 cells, even when they are differentiated with staurosporine.

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Section: Discussionmentioning

confidence: 87%

Sensitivity of Staurosporine-Induced Differentiated RGC-5 Cells to Homocysteine

Ganapathy

Dun

et al. 2009

Current Eye Research

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“…In case of the prostate cancer, LNCaP line (androgensensitive human prostate adenocarcinoma cells derived from the supraclavicular lymph node metastasis) proteomic characteristics of low (L-33) and high (H-81) passage numbers showed significant differences in protein expression (Youm et al 2008). Authors identified five proteins (Tim, cathepsin D, CKB, GRP78, HSP27) that exhibited significantly different changes between the low and high passage number.…”

Section: Discussionmentioning

confidence: 99%

Changes in cellular response to the damage induced in PC-3 prostate cancer cells by proton microbeam irradiation

Lipiec¹,

Wiecheć²,

Dulińska-Litewka³

et al. 2012

gpb

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The aim of this research was to find out whether the passage number effect may influence on the PC-3 cells (the human prostate cancer line derived from bone metastases) response to proton radiation. 2 MeV horizontally focused proton microbeam was used as a radiation source. The cells were treated with a counted number of H(+) ions (50-8000) corresponding to doses of 1.3-209 Gy/cell. For comparison, cell death was also induced by UVC radiation. All cells were stained with Hoechst 33342 and propidium iodide and visualized under a fluorescence microscope. Necrosis was observed at: a) 8000 protons per cell (corresponding to ∼209 Gy/cell) after 2-4 passages, b) 3200 protons per cell (corresponding to ∼84 Gy/cell) for cells after 11-14 passages and c) only 800 protons per cell (corresponding to ∼2 Gy/cell ) after 47-50 passages. Apoptosis was efficiently induced, by protons, only in cells after 50 passages. The results showed that the laboratory conditions affected cellular response of PC-3 cell line to the proton irradiation. The cellular response to the radiation treatment strongly depends on number of passages.

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“…It is also known that HSP27 plays important roles in the apoptosis signal transmission process related to caspase-3: HSP27 interrupts cytochrome c secretion of thread granules in the procaspase-9 pathway, and it inhibits apoptosis by interrupting caspase-3 activation and apoptosome formation by acting on cytochrome c or procaspase-3 [4]. One study showed that HSP expression inhibited apoptosis, and the other previous studies found that HSP27 was related to the hormone resistance acquisition of the LNCaP cell line, which is a kind of a prostate cancer [5,6]. …”

Section: Introductionmentioning

confidence: 99%

Expression of Heat Shock Protein 27 in Prostate Cancer Cell Lines According to the Extent of Malignancy and Doxazosin Treatment

Lee

Chung

et al. 2013

World J Mens Health

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PurposeHeat shock protein 27 (HSP27) is known as the material that plays a role in apoptosis control in tumor and cell protection including the immune response, drug tolerance, and so on. In this study, HSP27 expression according to the prostate cancer malignancy level was evaluated, and HSP27 expression was also analyzed after inducing apoptosis by doxazosin treatment of the prostate cancer cell lines.Materials and MethodsReverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining of the HSP27 was implemented by the culture of RWPE-1, LNCaP, androgen-independent human prostate cancer cells (PC-3), and TSU-Pr1 cell lines. Analysis was separately conducted in the control group, control vector group treated by dimethyl sulfoxide, and groups treated with 10 µM or 25 µM doxazosin. The expression of HSP27 in RT-PCR and immunofluorescence staining was observed and evaluated after conversion to numerical values.ResultsIn the RT-PCR results, depending on the cell type, LNCaP, TSU-Pr1 showed the highest HSP27 expression followed by PC-3, LNCaP and RWPE-1 in sequence. After doxazosin treatment, the expression detected by RT-PCR was stronger at a 25-µM doxazosin concentration compared to that at a 10-µM concentration, and the result was similar by immunofluorescence staining.ConclusionsHSP27 expression increased depending on the prostate cancer cell line. This meant that HSP27 expression was related to the prostate cancer malignancy level. Additionally, the higher the treatment concentration in PC-3 was, the higher the HSP27 expression was. This result showed that doxazosin induced apoptosis of prostate cancer.

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Proteomic analysis of androgen-independent growth in low and high passage human LNCaP prostatic adenocarcinoma cells

Cited by 7 publications

References 26 publications

Sensitivity of Staurosporine-Induced Differentiated RGC-5 Cells to Homocysteine

Sensitivity of Staurosporine-Induced Differentiated RGC-5 Cells to Homocysteine

Changes in cellular response to the damage induced in PC-3 prostate cancer cells by proton microbeam irradiation

Expression of Heat Shock Protein 27 in Prostate Cancer Cell Lines According to the Extent of Malignancy and Doxazosin Treatment

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