scholarly journals Altered staining patterns and expression level of Engrailed-2 in Benign prostatic hyperplasia and Prostate Cancer predict Prostatic disease progression

2019 ◽  
Author(s):  
Qi Li ◽  
Yibo Shi ◽  
Rigai Sa ◽  
Jun Hao ◽  
Jinhao Hu ◽  
...  

Abstract Background: Prostate cancer (PC) as a kind of malignant tumor, causes the most death of cancer among males. Successful curing of PC greatly relies on its diagnose in the early stage. Engrailed-2 (EN2), which has been confirmed being existed in the high level in the urine of PC patients, has not been reported as a histochemical diagnostic biomarker of PC. In this study, we analyzed the EN2 expression level and staining patterns in PC and benign prostatic hyperplasia (BPH) samples for seeing the change of EN2 in PC early stage. Methods: EN2 monoclonal antibody was generated and the specificity of this antibody was validated with different maneuvers. Endogenic and exogenous of EN2 in three PC cell lines (LNCap, PC3, and DU145) was detected by immunofluorescence. The expression level and staining patterns of EN2 in 25 of PC and 25 of BPH tissues were detected by immunohistochemistry. RT-PCR was done for further conforming whether EN2 is overexpressed in PC and BPH tissues. Finally, a relationship of EN2 expression and PC occurrence was obtained by case analysis of PC. Results: The results of WB and immunofluorescence showed the monoclonal antibody of EN2 we made could specifically bind endogenic and ectogenic EN2 protein in three different PC cell lines. Results of immunofluorescence showed the endogenic EN2 was generally expressed in the cytoplasm and ectogenic EN2 has mostly existed in the nucleus. Immunohistochemical staining of EN2 in PC was extremely higher than in BPH confirmed by RT-PCR. The staining areas were mostly nucleus and cytoplasm in BPH tissues but cytomembrane in PC tissues. The expression level of EN2 was positively correlated with the PC clinical stage. Conclusion: The EN2 monoclonal antibody we made could distinguish BPH and PC by immunohistochemistry. The expression level and tissue distribution pattern of EN2 can help with the determination of PC’s progression.

2020 ◽  
Author(s):  
Qi Li ◽  
Yibo Shi ◽  
Rigai Sa ◽  
Jun Hao ◽  
Jinhao Hu ◽  
...  

Abstract Background: Prostate cancer (PC) as a kind of malignant tumor, causes the most death of cancer among males. Successful curing of PC greatly relies on its diagnose in the early stage. Engrailed-2 (EN2), which has been confirmed being existed in the high level in the urine of PC patients. In this study, we determine if there were differences in the staining patterns and expression level of EN2 in benign prostatic hyperplasia (BPH) and PC.Methods: Immunohistochemical and RT-PCR analysis of the expression of EN2 was conducted in 25 PC and 25 BPH cases. EN2 monoclonal antibody against EN2 helix 3 was developed and its specificity was identified. The subcellular localization of endogenic and exogenous EN2 in three PC cell lines (LNCap, PC3, and DU145) was detected by immunofluorescence. Correlation among clinical indicators and EN2 immunohistochemical scores of these 25 PC and 25 BPH cases were analyzed and two representative PC cases with different EN2 expression were used to vividly illustrate the correlation between EN2 expression and PC clinical stage. Results: The results of western-blotting (WB) and immunofluorescence showed homemade EN2 monoclonal antibody could specifically bind endogenic and ectogenic EN2 protein in three different PC cell lines. Results of immunofluorescence showed the endogenic EN2 was generally expressed in the cytoplasm and ectogenic EN2 has mostly existed in the nucleus of three PC cell lines. Immunohistochemical staining of EN2 in PC was extremely higher than in BPH confirmed by RT-PCR. The staining areas were mostly nucleus and cytoplasm in BPH tissues but cytomembrane in PC tissues. The expression level of EN2 was positively correlated with the PC clinical stage. Conclusion: The EN2 monoclonal antibody we made could be used in immunohistochemistry to display the expression pattern of EN2 in BPH and PC. The staining patterns and expression level of EN2 in BPH and PC are different.


2020 ◽  
Author(s):  
Qi Li ◽  
Yibo Shi ◽  
Rigai Sa ◽  
Jun Hao ◽  
Jinhao Hu ◽  
...  

Abstract Background: Prostate cancer (PC) , a common malignant tumor, is the second-leading cause of cancer death among American men. Its successful treatment greatly relies on the early diagnose. Engrailed-2 (EN2) has been confirmed being existed with a high level in the urine of PC patients. In this study, to explore the application of EN2 in PC, we detected the immunohistochemical staining difference and EN2 expression level between benign prostatic hyperplasia (BPH) and PC. Methods: We developed a monoclonal antibody against the helix 3 in EN2 and confirmed its specificity with Western blotting (WB) and immunofluorescence detecting the subcellular localization of endogenous and exogenous EN2 in three PC cell lines (LNCap, PC3, and DU145). We conducted immunohistochemical staining using this homemade antibody, and RT-PCR to detect the expression of EN2 in 25 PC and 25 BPH cases , and analyzed the correlation of EN2 expression and PC clinical staging. Results: The results of WB and immunofluorescence showed our homemade EN2 monoclonal antibody could specifically bind endogenous and exogenous EN2 protein in three different PC cell lines. Endogenous EN2 was generally expressed in the cytoplasm and exogenous EN2 mostly existed in the nucleus of these cell lines. Immunohistochemical staining in PC had extremely stronger signals than that in BPH, suggesting a higher EN2 expression level in PC, which was confirmed by RT-PCR. Interestingly, the stained areas in BPH tissues were mainly in nucleus and cytoplasm, while in PC tissues were mainly on cytomembrane. Moreover, the expression level of EN2 was positively correlated with the PC clinical staging. Conclusion: Using our homemade EN2 antibody, we have found different staining patterns and expression level of EN2 in BPH and PC,which may be helpful to predict prostatic disease progression.


Author(s):  
Niloofar Dehghani ◽  
Masoud Salehipour ◽  
Babak Javanmard

Introduction: Prostate cancer is the second most common cancer and the leading cause of cancer-related deaths worldwide. In the present study, the expression level of glycine N-methyl transferase gene (GNMT) was investigated in prostate cancer tissue. The GNMT enzyme is encoded by the GNMT gene. Increased GNMT gene expression increases the conversion of glycine to sarcosine and results in the elevated levels of sarcosine in blood and urine. Methods: The expression level of GNMT gene in tissue samples of patients with prostate cancer was compared with those with benign prostatic hyperplasia using Real-Time PCR technique. Results: The GNMT gene expression level increased significantly in prostate cancer patients compared with those with benign prostatic hyperplasia (p-value <0.001). In addition, the expression level of GNMT gene was stage-dependent and  significant increases were observed in all stages of prostate cancer compared with those with benign prostatic hyperplasia (p-value <0.001). Conclusion: The concentration of sarcosine is controlled by GNMT and it seems that increasing the expression level of GNMT gene increases the level of sarcosine concentration. Thus, it appears that increased levels of GNMT expression occur in the early stages of prostate cancer. Therefore, periodic measurement of GNMT expression levels can detect prostate cancer before it forms a cancer cell and invades other tissues.


2008 ◽  
pp. 451-458
Author(s):  
D Housa ◽  
Z Vernerová ◽  
J Heráček ◽  
B Procházka ◽  
P Čechák ◽  
...  

Serum levels of adiponectin were measured in patients with benign prostatic hyperplasia and prostate cancer of pT2 and pT3 stage. Adiponectin ELISA assay, immunohistochemistry, and selected metabolic and biochemical parameters measurement was performed in 25 patients with benign prostatic hyperplasia and 43 with prostate cancer (17 patients with organ-confined and 26 patients with locally advanced disease). Serum adiponectin levels did not differ between prostate benign hyperplasia and cancer clinical stage T2, but was significantly higher in pT3 relative to pT2 group (14.51±4.92 vs. 21.41±8.12, P = 0.003). Tissue immunohistochemistry showed enhanced staining in neoplastic prostate glands and intraepithelial neoplasia relative to benign prostatic hyperplasia without distinction between disease grade and stage. Serum adiponectin levels are higher in locally advanced relative to organ-confined prostate cancer and may thus serve as an auxiliary marker providing further improvement for discrimination between pT2 and pT3 stages.


2004 ◽  
Vol 171 (4S) ◽  
pp. 125-125
Author(s):  
Lizhong Wang ◽  
Kazunari Sato ◽  
Norihiko Tsuchiya ◽  
Chikara Ohyama ◽  
Shigeru Satoh ◽  
...  

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.


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