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Inflammation is associated with promotion of the initiation of various malignancies, partly due to bacterial infection-induced microenvironmental changes. However, the exact association between microbiota in urine, seminal fluid and the expressed prostatic secretions and benign prostatic hypertrophy and prostate cancer is not clear.
In the present study, we investigated the type of microbiota in the expressed prostatic secretions EPS of patients with prostate cancer and benign prostatic hyperplasia BPH by the polymerase chain reaction-denaturing gradient gel electrophoresis PCR-DGGE method using universal bacterial primers. In order to understand the possible association between various Prostatitis und Helicobacter and prostate cancer, quantitative real-time PCR assay was performed to quantify the amount of strains of bacteria in urine, EPS and seminal fluid.
The prostate cancer group had a significantly increased number of Bacteroidetes bacteria, Prostatitis und Helicobacter, Firmicutes bacteria, Lachnospiraceae, PropionicimonasSphingomonasand Ochrobactrumand a decrease in Eubacterium and Defluviicoccus Prostatitis und Helicobacter to the BPH group.
The number of Escherichia coli in the prostate cancer group was significantly decreased in urine and increased in the EPS and seminal fluid, while the number of Enterococcus was significantly increased in the seminal fluid with little change in urine and EPS. Based on these results, we suggest that there are significant changes in the microbial population in EPS, urine and seminal fluid of subjects with prostate cancer and BPH, indicating a possible role for Prostatitis und Helicobacter bacteria in these two conditions.
Prostate cancer is one of the most common cancers in men, especially in the elderly. It has long been suspected that chronic inflammation may have an important role in the initiation and progression of prostate cancer [ 1 — 3 ]. The observation that Helicobacter pylori induces chronic inflammation and thus may pave Prostatitis und Helicobacter way to develop stomach cancer [ 3 ] lends support to such a belief that bacteria may also have Prostatitis und Helicobacter role in prostate cancer [ 3 ].
There is some support to this contention that prostatic inflammation may increase the risk of benign prostatic hyperplasia BPH and prostate cancer [ 45 ]. It is difficult to detect many anaerobic bacteria present in various human body fluids, especially in the urine, expressed prostatic secretions EPS and Prostatitis und Helicobacter fluid, using traditional culture methods [ 67 Prostatitis und Helicobacter. Hence, in the present study, we employed culture-independent methods polymerase chain reaction-denaturing gradient gel electrophoresis — PCR-DGGE that are more reliable to detect various bacteria present in the human body fluids [ 8 ].
This method has previously been used by others to identify a variety of bacteria in various secretion of the human body [ 8 — 12 ]. Gram-negative enteric bacteria including Escherichia coliPseudomonas aeruginosaand Enterococcus are frequently associated with urinary tract infections that may cause chronic bacterial prostatitis [ 13 ].
Furthermore, infections due to E. Hence, it is reasonable to propose that chronic bacterial prostatitis induced by bacteria may enhance proinflammatory responses that may contribute to prostate cancer. All male patients included in the study were Prostatitis und Helicobacter the age of 75 and diagnosed with BPH or prostate cancer. All patients were educated as to how to collect their urinary samples without contamination. Expressed prostatic secretions and seminal fluid were collected following prostatic massage in a 5.
Urine, EPS and seminal fluid were carefully collected into the tubes without touching the interior wall of the sterile tube. Routine urine test was performed before sample storage.
Following brief centrifugation, the mixture was carefully Prostatitis und Helicobacter to a QIAamp Mini spin column without wetting the rim. An additional 1-min-long centrifugation at rpm was performed and the column was placed in a clean 2 ml collection tube. Subsequently the column was kept in a clean 1. DNA was collected from the columns by centrifuging at rpm for 1 min.
The protocol for the PCR procedure was the touchdown protocol as described by Muyzer et al. Based on the similarity search results, a phylogenetic tree was constructed by the neighbor-joining method Prostatitis und Helicobacter the MEGA 5 software version 5. The consistency of the relationship on the tree was assessed by bootstrap resampling. Bootstrapping was performed for trials in accordance with the Clustal X program [ 19 ].
A DNA sample with the Prostatitis und Helicobacter concentration was diluted 10, 10 210 3 Prostatitis und Helicobacter, 10 410 5 times and amplified under the same PCR conditions for the standard curves.
Statistical analysis was performed by analysis of variance or by paired t -test when just two values were compared, using SPSS software version 9. The mean Prostatitis und Helicobacter of the recruited subjects in both groups was Urine and blood analysis was performed before specimen collection.
The characteristics of the tumor markers of patients with prostate cancer studied and their results are given in Table III. Compared to patients with prostatic hyperplasia, patients with prostate cancer had higher incidence of occult blood and turbidity, and a higher quantity of white blood count WBC and bacteria in the urine, suggesting that these patients might be having concurrent infection and inflammation that could be due to the presence of infection of the prostate or urethritis.
Three inferences could be arrived at from these findings. They are: i active infection is present in the prostate at the time of the study; ii active infection is present in the urinary tract; and iii active infection is present in both the prostate and the urinary tract.
CEA — carcino-embryonic, CA — carbohydrate antigen Based on the results obtained, it is evident that 16s RNA gene fragments of bacteria in the specimens had good amplification efficiency.
Polymerase chain Prostatitis und Helicobacter gradient gel electrophoresis fingerprinting analysis of patients with BPH and prostate cancer for predominant bacteria was used to identify EPS microbiota Figure 3. Based on PCR-DGGE fingerprinting analysis, the diversity of microbiota structures was presented with separated bands of different DNA sequences, as shown in Figure 3which showed that there were about 20 discernible bands.
Gel profiles of the V3 region amplified by polymerase chain reaction from bands 5, 6, 8, 10, 13, 15, 16, 17 and These distinct bands, namely 5, 6, 8, 10, 13, 15, 16, 17 and 19, were identified as significant variables in discriminating the prostate cancer group from the BPH group.
These distinct bands Prostatitis und Helicobacter sequenced to determine the exact microorganisms that were present in the urinary system during prostate cancer.
The bands 5, 6, 8, 13, 16, 17, and 19 that were brighter in the prostate cancer and bands 10 and 15 in the BPH groups were amplified, and the lengths of these fragments were about bp Figure 4. Prostatitis und Helicobacter sequences of prominent bands 5, 6, 8, 13, 16, 17, and 19 in the prostate cancer group matched with Bacteroidetes bacteria, Alphaproteobacteria, Firmicutes bacteria, Lachnospiraceae, PropionicimonasSphingomonasOchrobactrum and bands 10 and 15 of BPH matched with Eubacterium and Defluviicoccus species respectively.
It is evident from these results that there were almost no significant differences in the diversity of bacteria between the groups except for the fact that seven kinds of microbes in the prostate cancer group were distinctly increased in quantity compared with those in the BPH group. However, Eubacterium and Defluviicoccus were present in lower amounts in the prostate cancer group compared with the BPH group.
Among these strains, the one that showed the highest similarity of the same category was selected. Based on these results, the phylogenetic tree was created by MEGA 5. As shown in the phylogenetic tree Figure 5most of the strains were uncultured and values on Prostatitis und Helicobacter branches of the tree showed a high confidence coefficient and close genetic relationships. The standard curves of both Prostatitis und Helicobacter PCR assays E. The amount of E. The qPCR results of E.
Previously, it was reported that prostatitis and prostatitis symptoms are associated with an increased risk of prostate cancer [ 5 ]. It is likely that bacteria might induce a chronic inflammatory state in the prostate that results in enhanced production of pro-inflammatory cytokines. Both neutrophils and macrophages may release proinflammatory molecules such as nitric oxide that Prostatitis und Helicobacter the propensity to cause genetic damage that could pave the way for enhanced cell proliferation and cancer.
This is particularly interesting in the light of the reports that low-grade inflammation exists in cancer. Some studies did show that specific types of bacterial sequences could be present in prostatic Prostatitis und Helicobacter that may not be detected by traditional cell culture methods [ 20 ].
Molecular-based methods to identify and characterize microorganisms have contributed to microbial discovery. Some previous studies evaluated bacterial 16S rDNA sequences in prostatic tissue from patients with prostate cancer [ 23 ]. In the present study, using a combination of PCR-DGGE fingerprinting analysis and sequencing, combined with phylogenetic analysis, we observed that the microbiota present in the urinary system are significantly different between patients with BPH and prostate cancer.
Denaturing gradient gel electrophoresis fingerprints indicated that Prostatitis und Helicobacter urinary microbiota compositions in the EPS of prostate cancer were similar to those seen in BPH though there were significant differences in the quantities of specific strains between the groups.
The sequencing results showed that EPS of prostate cancer patients Prostatitis und Helicobacter rich in Bacteroidetes bacteria, Alphaproteobacteria, Firmicutes bacteria, Lachnospiraceae, PropionicimonasSphingomonasand Ochrobactrumwhich might be involved in the progression of prostate cancer. Capsular polysaccharides produced by Bacteroidetes bacteria are important pathogenic factors that can cause suppurative lesions in the abdominal cavity and other organs.
Sphingomonas is a genus of Gram-negative bacteria that belongs to the Alphaproteobacteria, which is one of the commonest groups associated with urinary tract infection.
Hence, increase of Alphaproteobacteria Prostatitis und Helicobacter 6 and 17 in the prostate cancer group may be relevant to the infection and inflammation of the prostate seen in patients with prostate cancer. Firmicutes bacteria account for the Prostatitis und Helicobacter proportion of the human gut microbiome and are known to be involved in energy resorption and obesity [ 24 ].
The number of Firmicutes in vertebrates including humans is considered to be diet-dependent and correlates with caloric intake. Since a high-fat and high-caloric diet is widely believed to be an important factor involved in the genesis of prostate cancer, it is reasonable to propose that the increased number of Firmicutes band 8 and 13 seen in the EPS of patients with prostate cancer may have a role in its pathogenesis.
Propionicimonas belongs to the Propionibacterineae, which are the main anaerobic bacteria in EPS of prostatitis patients. Ochrobactrum is an opportunistic pathogen. In the present study, we observed that the quantity of Ochrobactrum in the EPS of prostate cancer patients was much higher compared with BPH, which suggests the immune dysfunction of these patients. This implies that efforts to suppress the growth of Ochrobactrum could be of significant benefit in the prevention of prostate cancer development.
The results of the present investigation showed that E. Increased levels of these two types of bacteria in EPS and seminal fluid indicate that a significant degree of inflammation occurs in those with prostate cancer.
Several previous studies have used prostate cancer tissue to detect the presence and Prostatitis und Helicobacter of bacteria in prostate cancer [ 232526 ]. In contrast, we collected EPS to investigate the presence and possible shift in the type and quantity of microbiota in BPH and prostate cancer.
Expressed prostatic secretions are secreted by the prostate and pass through the urethra, which is connected to the skin. Hence, it is likely that bacteria in the male reproductive tract and urethra could access EPS, and so their analysis Prostatitis und Helicobacter more likely reflect a shift in the type of bacteria present in the prostate with different diseases.
These results suggest that Prostatitis und Helicobacter dysbiosis of the bacterial community in the EPS might play an important role in the pathobiology of prostate cancer. National Center for Biotechnology InformationU. Journal List Arch Med Sci v.
Arch Med Sci. Published online Apr Das 4. Undurti N. Author information Article notes Copyright and License information Disclaimer. Corresponding author. Corresponding author: Dr. Phone: 86 E-mail: nc. E-mail: moc. This article has been cited by other articles in PMC. Abstract Introduction Inflammation is associated with promotion of the initiation of various malignancies, partly due to bacterial infection-induced microenvironmental changes.
Material and methods In the present study, we investigated the type of microbiota in the Prostatitis und Helicobacter prostatic secretions EPS of patients with prostate cancer and benign prostatic hyperplasia BPH by the polymerase chain reaction-denaturing gradient gel electrophoresis PCR-DGGE method using universal bacterial primers.
Results The prostate cancer group had a significantly increased number of Bacteroidetes bacteria, Alphaproteobacteria, Firmicutes bacteria, Lachnospiraceae, PropionicimonasSphingomonasand Ochrobactrumand a decrease in Eubacterium and Defluviicoccus compared to the BPH group. Conclusions Based on these results, we suggest that there are significant changes in the microbial population in EPS, urine and seminal fluid of subjects with prostate Prostatitis und Helicobacter and BPH, indicating a possible role for these bacteria in these two conditions.