Phenotypic and genotypic characterization of commonly used human prostatic cell lines

Abstract

Objective To investigate and catalogue systematically the phenotypic and genotypic characteristics of the commonly used prostatic cell lines using immunocytochemistry and polymerase chain reaction (PCR) of hypervariable sequences within the genome to provide a ‘fingerprint’ characteristic of each cell line. Materials and methods Malignant (LNCaP, LNCaP‐r, PC‐3, DU‐145) and benign immortalized prostatic cell lines (PNT‐1A, PNT‐1B, BPH‐1) were grown on four‐well slides, fixed and subjected to indirect streptavidin‐biotin immunocytochemistry. Twenty‐three antibodies were used in the following groups: cytoskeletal elements: cytokeratins (CK)‐5, ‐7, ‐8, ‐14 (two), ‐16, ‐18, ‐19 (three), ‐20, vimentin and desmin; MUC1 (three); cell adhesion molecules (E‐cadherin, α‐β‐and γ‐catenin); and prostatic associated proteins: prostate specific antigen (PSA), prostatic acid phosphatase (PAP) and androgen receptor (AR). For the PCR, genomic DNA was extracted from the cell lines and from SKOV3 and MCF7 (positive controls). PCR was performed on three variable regions which were then sequenced: AR exon 1 (CAG repeat polymorphism), and two areas of microsatellite instability (MSI): AR exon 8 and hypoxanthine‐guanine phosphoribosyl transferase (HPRT) exon 3. Results All cell lines were CK‐8/18 positive and most also expressed CK‐7 and ‐19. Heterogeneous CK‐20 expression was detected for the first time in prostatic cell lines. All lines were positive for vimentin and negative for desmin. MUC1 was expressed in one malignant (DU‐145) and all immortalized cell lines. E‐cadherin expression was low or absent in three lines: PNT1A, 1B and PC‐3. Only PC‐3 failed to express α‐catenin; β‐ and γ‐catenin were expressed by all lines. PSA, PAP and AR were only expressed by LNCaP and LNCaP‐r. On PCR, the CAG repeat lengths in exon 1 of the AR ranged from 19 to 27. Three pairs of cell lines had the same exon 1 CAG repeat length: LNCaP/PC‐3 (26 repeats), BPH‐1/DU‐145 (19 repeats) and PNT1 A/1B (20 repeats). Exon 8 sequences were identical except for LNCaP, which showed a single base mutation, and HPRT exon 3 sequences were all identical. There was no evidence of generalized MSI in any of the cell lines examined. Conclusions The cell lines studied fell into three broad groups according to their phenotypic characteristics: (i) prostatic marker positive (LNCaP and LNCaP‐r); (ii) high expression of most antigens (DU‐145, PC‐3 and BPH‐1); and (iii) low or absent expression of most antigens (PNT1 A and 1B). Each of the cell lines derived from PC could be identified on the basis of exon 1 and 8 AR sequence variability. DU145 and BPH‐1 had identical profiles of the three areas studied, but these cell lines are easily distinguished by their different phenotypic characteristics. PNT1A and 1B had identical genetic and similar phenotypic profiles, which is unsurprising given that they are subclones derived from the same parental line. Even so, these were separable on the basis of CK‐19 immunostaining. Using a combination of geno‐ and phenotypic markers it was...

MUC1 expression in human prostate cancer cell lines and primary tumors

“…Of the five prostate cancer cell lines evaluated, DU-145 cells had the highest basal expression of MUC1. Our results with 28 who reported that unstimulated PC-3 cells were negative for MUC1 protein expression, whereas we found low basal MUC1 core protein expression in PC-3 cells. The most likely explanation for this discrepancy is that our use of MUC1 antibodies (214D4 and CT-1) not sensitive to glycoform variations, that is, more broadly recognizing MUC1, provides a more reliable method of revealing MUC1 protein expression.…”

“…[5][6][7] In this study, basal MUC1 protein expression was evaluated in normal prostate epithelial cells (PrEC) and a series of metastasis-derived prostate cancer cell lines, and was consistent with one previous report. 28 LNCaP cells and sublines, representing a model for progression to androgen independence and skeletal metastasis, 29 were negative for MUC1 expression, suggesting that the lack of MUC1 expression is a characteristic of LNCaP cells passed onto the sublines. It should be noted that genetic changes known to occur in LNCaP and sublines did not include the locus for MUC1.…”

Membrane androgen receptor activation in prostate and breast tumor cells: Molecular signaling and clinical impact

“…Interestingly, while LNCaP prostate cancer cells express functional intracellular androgen receptors (iAR), the DU145 cell line expresses either nonfunctional iAR (35), or is iAR-deficient (36,37). Therefore, DU145 cells fail to respond to iAR-regulated androgen treatment (35).…”

Rapid Molecular and Morphological Responses of Prostate Cell Lines to Cell–Cell Contact

“…α-, β-, and γ -catenins: α-catenin in PZ-HPV-7 cells was distributed around the rim of the cells, resulting in what Mitchell et al (2000) has described as a perinuclear halo, while less staining of the cytoplasm was observed (Figure 3i and ii c, arrow). β-and γ -catenins were distributed in small discrete intracellular regions 1-2 µm from the cell surface, in a pattern that led to the formation of an intracellular ring (Figure 3i and ii d, e, arrows).…”

“…The temporal development of the distribution of the cadherin/catenin complex in prostate or other cancer cell lines where that complex is impaired has not previously been studied. A genotypic and phenotypic analysis has been previously conducted on many prostate cell lines, with the purpose of providing meaningful comparison between similar cell lines held in different laboratories (Mitchell et al 2000). Of these, epithelial 'normal' (PZ-HPV-7) and cancer (DU-145) cell lines were selected for study in the present work.…”