mRNA expression analysis of human kallikrein 11 (KLK11) may be useful in the discrimination of benign prostatic hyperplasia from prostate cancer after needle prostate biopsy

Kallikrein 11 (KLK11, TLSP, hippostatin) is a member of the human kallikrein gene family, which includes PSA, KLK2 and 12 other members, all localized on chromosome 19q13.4. The aim of this study was to investigate whether KLK11 expression could be used to discriminate prostate cancer (CaP) from benign prostatic hyperplasia (BPH) in needle prostate biopsies. We analyzed the expression of the prostate-type variant of the KLK11 gene in 64 CaP and BPH tissues obtained by transrectal ultrasound-guided needle biopsy. Reverse transcription (RT), PCR and image analysis methodologies were developed and used. Of the 42 BPH tissues examined, only 10 (23.8%) were positive for prostate-type KLK11 expression, while of the 22 CaP patients, 12 (54.5%) were KLK11-positive (p=0.025). Logistic regression and receiver operating characteristic curve analyses demonstrated that KLK11 expression has a significant discriminatory value (crude odds ratio=3.84, p=0.016; area under the curve, 0.65, 95% CI 0.51-0.80) between CaP and BPH in needle prostate biopsies. Copyright © by Walter de Gruyter

Quantitative analysis of human kallikrein 5 (KLK5) expression in prostate needle biopsies: An independent cancer biomarker

BACKGROUND: Kallikrein 5 (KLK5), a recently cloned member of the kallikrein family, codes for the secreted protein KLK5. Active KLK5 protein has a trypsin activity, and the expression of KLK5 gene seems to be regulated by steroid hormones. We performed an expression analysis and clinical evaluation of the KLK5 gene, at the mRNA level, in prostate needle biopsies. METHODS: We examined KLK5 mRNA concentrations in 103 prostate tissue specimens. After testing of RNA quality, cDNA was prepared by reverse transcription.A highly sensitive quantitative real-time PCR (qRTPCR) method for KLK5 mRNA quantification was developed using the SYBR Green chemistry. GAPDH was used as a housekeeping gene. RESULTS: Specimens from patients with benign prostatic hyperplasia (BPH) showed higher levels of KLK5 mRNA expression than those from patients with prostate cancer (PCa) (P = 0.024). ROC analysis demonstrated that KLK5 expression had significant discriminatory value between BPH and PCa (AUC 0.64; P = 0.016). KLK5 mRNA expression showed a statistically significant negative correlation with the total PSA serum concentration in the PCa patients (P = 0.003). Early-stage tumors showed higher KLK5 expression than late-stage ones (P=0.014), whereas KLK5 expression was negatively correlated to Gleason score (P = 0.005). CONCLUSIONS: KLK5 mRNA, analyzed by quantitative PCR in prostate needle biopsies, could be an independent biomarker for the differential diagnosis and prognosis in prostate cancer. © 2009 American Association for Clinical Chemistry

Expression analysis and prognostic significance of human kallikrein 11 in prostate cancer

Background: Kallikrein 11 (KLK11) is a newly discovered human kallikrein gene that is mainly expressed in the central nervous system and endocrine tissues. KLK11 has two alternative splicing isoforms, known as the brain type and prostate type. Many members of the human kallikrein gene family are differentially expressed in cancer and a few have potential as diagnostic/prognostic markers. Methods: In the present study, the expression of prostate type variant of KLK11 gene was analyzed by RT-PCR in 66 prostate cancer tissues. Tumors were pulverized, total RNA was extracted, and cDNA was prepared by reverse transcription. KLK11 was amplified by PCR using gene specific primers and its identity was verified by sequencing. Prostate tissues were then classified as KLK5 positive or negative based on eithidium bromide staining in agarose gels and image analysis. Results: KLK11 was found to be highly expressed in 43/66 (65%) of prostate cancer samples. We found a significant negative relationship between KLK11 expression and Gleason score (p = 0.004) and disease stage (p = 0.038). Serum total PSA concentration was found to be lower in patients with overexpression of KLK11 (p = 0.044). Conclusions: We conclude that down-regulation of the KLK11 gene in advanced and more aggressive tumors may open the possibility of being used as a future biological marker distinguishing the tumor aggressiveness as well as a useful prognostic biomarker for prostate cancer. © 2005 Elsevier B.V. All rights reserved

Expression analysis and prognostic significance of human kallikrein 11 in prostate cancer

Background: Kallikrein 11 (KLK11) is a newly discovered human kallikrein gene that is mainly expressed in the central nervous system and endocrine tissues. KLK11 has two alternative splicing isoforms, known as the brain type and prostate type. Many members of the human kallikrein gene family are differentially expressed in cancer and a few have potential as diagnostic/prognostic markers. Methods: In the present study, the expression of prostate type variant of KLK11 gene was analyzed by RT-PCR in 66 prostate cancer tissues. Tumors were pulverized, total RNA was extracted, and cDNA was prepared by reverse transcription. KLK11 was amplified by PCR using gene specific primers and its identity was verified by sequencing. Prostate tissues were then classified as KLK5 positive or negative based on eithidium bromide staining in agarose gels and image analysis. Results: KLK11 was found to be highly expressed in 43/66 (65%) of prostate cancer samples. We found a significant negative relationship between KLK11 expression and Gleason score (p = 0.004) and disease stage (p = 0.038). Serum total PSA concentration was found to be lower in patients with overexpression of KLK11 (p = 0.044). Conclusions: We conclude that down-regulation of the KLK11 gene in advanced and more aggressive tumors may open the possibility of being used as a future biological marker distinguishing the tumor aggressiveness as well as a useful prognostic biomarker for prostate cancer. (c) 2005 Elsevier B.V. All rights reserved

Free/Total PSA (F/T ratio) kinetics in patients with clinically localized prostate cancer undergoing radical prostatectomy

Background: In this paper we study the Free/Total PSA kinetics in patients with clinically localized prostate cancer undergoing radical prostatectomy. Methods: Serum PSA, Free PSA and Free/Total Ratio were determined preoperatively, at the time of prostate removal (0 time) and then at 3, 6, 12, 24, 48, 72 and 168 h, from 9 patients with clinically localized prostate cancer who underwent radical retropubic prostatectomy. The elimination rates and half-lives of Total, Free PSA and F/T Ratio were studied applying one and two compartment models for pharmacokinetic analysis. Results: Surgical manipulations of the prostate caused a mean 2.16-fold increase of PSA, 12-fold increase of free PSA and 4.2-fold increase of F/T PSA ratio. Removal of the prostate caused a rapid biphasic, biexponential elimination of Free PSA with a mean half-life of 0.8 h for the alpha (a) phase and 32.6 h for the beta (b) phase. PSA was eliminated following a rapid exponential (a) phase with a half-life of 1.15 h and a non-exponential (b) phase with a half-life of 71.96 h. Free/Total PSA followed a biphasic kinetic, with an initial exponential elimination phase and a mean half-life of 2.6 h and a second non-exponential increase phase with a doubling time of 130.8 h. Free/Total PSA reached its nadir very soon, at the first postoperative 24 h. Conclusions: Free/Total PSA kinetic after radical prostatectomy reflects the differences of Free and Total PSA elimination kinetics. Free/Total Ratio follows a biphasic kinetic, with an initial rapid exponential elimination phase, which is affected mainly by the rapid exponential (a) phase of Free PSA elimination and a second slow increase, which is affected mainly by the terminal non-exponential (b) phase of PSA elimination. © 2005 Elsevier B.V. All rights reserved

Total and free PSA kinetics in patients without prostate cancer undergoing radical cystoprostatectomy

BACKGROUND. Radical cystoprostatectomy and radical prostatectomy are the two major operations where prostate is totally and radically removed. Radical cystoprostatectomy is usually performed in patients with invasive bladder cancer. The aim of the study was to examine Total PSA, Free PSA, and Free/Total Ratio elimination kinetics after radical cystoprostatectomy. METHODS. Serum PSA, Free PSA, and Free/Total Ratio were determined preoperatively, at the time of cystoprostatectomy specimen removal and then at 3, 6, 12, 24, 48, 72, and 168 hr, from seven patients with muscle invasive bladder cancer, who underwent radical cystoprostatectomy. Free and Total PSA concentrations were measured with non-competitive immunological procedures. The elimination rates and half-lives of Total, Free PSA and Free/Total Ratio were studied using a nonlinear regression analysis. RESULTS. Surgical manipulations caused about 1.5-fold increase of PSA, 5-fold increase in Free PSA and 3-fold increase in Free/Total Ratio. PSA and Free PSA followed a biphasic elimination pattern of a rapid exponential (a) phase with a half-life of 4.27 and 2.14 hr and a terminal, nonexponential (b) phase with a half-life of 63 and 173.2 hr, respectively. Free/Total PSA Ratio followed, also, a biphasic kinetic pattern of a rapid exponential decline with a half-life of 3.34 and a terminal non-exponential increase with a doubling time of 43 hr. CONCLUSIONS. Comparing PSA kinetics after radical cystoprostatectomy with those of radical prostatectomy, it appears that PSA follows the same elimination pattern in both models. In contrast, Free PSA and Free/Total Ratio elimination kinetics' patterns differ between the two surgical models. © 2008 Wiley-Liss, Inc

L-DOPA decarboxylase mRNA levels provide high diagnostic accuracy and discrimination between clear cell and non-clear cell subtypes in renal cell carcinoma

Objectives: Renal cell carcinoma (RCC) is the most frequent type of kidney cancer. RCC patients frequently present with arterial hypertension due to various causes, including intrarenal dopamine deficiency. L-DOPA decarboxylase (. DDC) is the gene encoding the enzyme that catalyzes the biosynthesis of dopamine in humans. Several studies have shown that the expression levels of DDC are significantly deregulated in cancer. Thus, we herein sought to analyze the mRNA levels of DDC and evaluate their clinical significance in RCC. Design and methods: DDC levels were analyzed in 58 surgically resected RCC tumors and 44 adjacent non-cancerous renal tissue specimens via real-time PCR. Relative levels of DDC were estimated by applying the 2-δδCT method, while their diagnostic accuracy and correlation with the clinicopathological features of RCC tumors were assessed by comprehensive statistical analysis. Results: DDC mRNA levels were found to be dramatically downregulated (. p<. 0.001) in RCC tumors, exhibiting remarkable diagnostic accuracy as assessed by ROC curve analysis (AUC: 0.910; p<. 0.001) and logistic regression (OR: 0.678; p=. 0.001). Likewise, DDC was found to be differentially expressed between clear cell RCC and the group of non-clear cell subtypes (. p=. 0.001) consisted of papillary and chromophobe RCC specimens. Furthermore, a statistically significant inverse correlation was also observed when the mRNA levels of DDC were analyzed in relation to tumor grade (. p=. 0.049). Conclusions: Our data showed that DDC constitutes a highly promising molecular marker for RCC, exhibiting remarkable diagnostic accuracy and potential to discriminate between clear cell and non-clear cell histological subtypes of RCC. © 2015