Background
Methods
Tissue samples
Immunohistochemistry
Quantitative Real-Time PCR
Gene | Forward primer | Reverse primer |
---|---|---|
GAPDH
| CCCCGGTTTCTATAAATTGAGC | CACCTTCCCCATGGTGTCT |
ACTB
| CCAACCGCGAGAAGATGA | CCAGAGGCGTACAGGGATAG |
PTGER1
| ATGGTGGTGTCGTGCATCT | CGCTGCAGGGAGGTAGAG |
PTGER2
| CCACCTCATTCTCCTGGCTA | AGGTCCCATTTTTCCTTTCG |
PTGER3
| ATCATGTGCGTGCTGTCG | TGCAGTGCTCAACTGATGTCT |
PTGER4
| CTCCCTGGTGGTGCTCAT | GGCTGATATAACTGGTTGACGA |
PTGFR
| GGGATCTACAGCCAGACCAG | GTGTCCTATGTCCTCCAAACG |
Statistical analysis
Results
Immunohistochemistry
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PTGER1 was expressed in most samples, but expression was increased in urothelial compared to smooth muscle cells. Most samples demonstrated high PTGER1 expression levels in urothelial cells, whereas muscle cells were expressing PTGER1 at moderate levels. Further analysis of PTGER1 expression profiles depicted comparable expression profiles in the renal pelvis, as well as the lower, mid and proximal ureter.
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PTGER2 expression was less prevalent: Urothelial cells demonstrated limited staining, irrespective of the localization in the examined ureter. In comparison, PTGER2 expression profiles showed weak staining of muscle cells in 15-20% of the ureter samples. PTGER2 expression was shown to be undetected in muscle cells located in the renal pelvis.
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PTGER3 was rarely detected in urothelial cells (10%) of the renal pelvis and the distal and proximal ureter. In comparison, PTGER3 was more prevalent in the smooth muscle cells, with different expression profiles, depending on the ureter localization: Expression profiles were lower in the upper parts of the examined ureters. In total, 62% of the distal ureter, 80% of the mid ureter, 46% of the proximal ureter and 23% of the renal pelvis samples expressed PTGER3 at low or moderate levels.
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PTGER4 expression was low, and detected more frequently in urothelial cells than in muscle cells. Only 13% of distal ureter muscle cells were expressing PTGER4 at low levels. PTGER4 expression was also low in urothelial cells, but detected in the distal ureter (23%), mid ureter (14%), proximal ureter (33%) and renal pelvis (31%).
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PTGFR expression was detected in most of the ureter samples: Less than 15% of the samples had undetectable PTGFR levels. The expression of PTGFR was similar in urothelial and smooth muscle cells.
Real-Time PCR
PTGER1 | PTGER2 | PTGER3 | PTGER4 | PTGFR | ||||||
---|---|---|---|---|---|---|---|---|---|---|
PCR | IHC | PCR | IHC | PCR | IHC | PCR | IHC | PCR | IHC | |
A-distal | 1.00 | +++ | 1.00 | ++ | 1.00 | - | 1.00 | + | 1.00 | +++ |
A-middle | 1.44 | ++ | 0.31 | + | 0.37 | +++ | 1.27 | - | 0.34 | ++ |
A-proximal | 3.15 | +++ | 0.62 | + | 0.42 | - | 1.09 | + | 0.64 | ++ |
A-renal pelvis | 2.73 | ++ | 0.55 | - | 0.36 | 1.63 | 0.55 | |||
B-distal | 0.61 | ++ | 0.14 | ++ | 0.69 | ++ | 0.67 | - | 0.31 | + |
B-middle | 0.41 | ++ | 0.22 | ++ | 0.95 | ++ | 0.87 | - | 0.31 | ++ |
B-proximal | 0.21 | ++ | 0.20 | + | 1.04 | ++ | 0.83 | - | 0.30 | +++ |
B-renal pelvis | 1.09 | ++ | 0.30 | + | 0.76 | + | 1.00 | 0.33 | ||
C-distal | 1.28 | +++ | 0.46 | ++ | 0.73 | ++ | 0.81 | - | 0.59 | +++ |
C-proximal | 1.11 | ++ | 0.36 | ++ | 1.23 | ++ | 0.84 | - | 0.64 | ++ |
C-renal pelvis | 3.51 | + | 0.29 | + | 2.14 | + | 1.80 | 0.49 |