GLI-1 is involved in EGF-regulated enhancement of the invasiveness of prostate cancer ARCaP(E) cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role of the hedgehog (HH) signaling pathway transcription factor glioma-associated oncogene hoinolog 1 (GLI-1) in EGF-regulated enhancement of the invasiveness of the prostate cancer ARCaP(E) cell line in vitro.</p><p><b>METHODS</b>The expressions of EGFR and GLI-1 in prostate cancer ARCaP(E) cells were analyzed by immunofluorescence staining. ARCaP(E) cells were treated with EGF at 100 ng/ml, followed by detection of the changes in cell morphology and invasiveness, as well as in the expressions of p-ERK, ERK and GLI-1. Migration transwell assay was used to determine the effects of 100 ng/ml EGF and GLI-1 antagonist GANT61 on the invasiveness of the ARCaP(E) cells.</p><p><b>RESULTS</b>Both EGFR and GLI-1 were expressed in the ARCaP(E) cells. EGF induced morphological transition of epithelial-like ARCaP(E) cells to mesenchymal-like cells, increased their in vitro invasiveness, and significantly upregulated the expressions of p-ERK and GLI-1 in the ARCaP(E) cells (P<0.05). GANT61 significantly inhibited the in vitro invasiveness of the ARCaP(E) cells and reduced the enhancing effect of EGF on their invasiveness (P<0.05).</p><p><b>CONCLUSION</b>The results from ARCaP(E) cells shed light on the cross-talk of the HH pathway with the EGF/ERK signaling pathway. GLI-1 might be responsible for EGF-regulated enhancement of the invasiveness of ARCaP(E) cells in vitro.</p>

Epithelial-mesenchymal transition and human fetal prostate development / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role and significance of epithelial-mesenchymal transition (EMT) and its transcriptional regulator Twist1 in the development of the human fetal prostate.</p><p><b>METHODS</b>Twenty-five human fetal prostate specimens at various developmental stages (16-39 weeks) were included in this study. EMT markers, such as E-Cadherin, N-Cadherin and Vimentin, and EMT transcriptional regulator Twist1 were determined by immunohistochemistry, and their relationship with the development of the human fetal prostate was analyzed.</p><p><b>RESULTS</b>E-Cadherin was expressed in the fetal prostate epithelium only, while Vimentin, N-Cadherin and Twist1 in both the epithelium and the stroma. The expression of E-Cadherin gradually increased, but those of Vimentin, N-Cadherin and Twist1 gradually decreased with the gestation stages. No significant changes were observed in the staining patterns of Vimentin, N-Cadherin and Twist1 in the stroma during the whole developmental process.</p><p><b>CONCLUSION</b>EMT is involved in the development of the human fetal prostate, which may promote epithelial cell motility to form prostatic bud tubules in early gestation stages and boost the differentiation of prostate epithelia in later stages.</p>

Comparison of transcription factors repressing epithelial phenotype in two different prostate cancer EMT models and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To screen and compare the specific transcription factors that repress the epithelial phenotype in epithelial-mesenchymal transition (EMT) in two different human prostate cancer models LNCaP/HIF1alpha and ARCaP.</p><p><b>METHODS</b>We established two different prostate cancer EMT models, LNCaP/HIF1alpha and ARCaP, cultured LNCaP, LNCaP/HIF1alpha, IF11 and IA8 cells in vitro, and detected the five transcription factors Snail, Slug, ZEB1, SIP1 and Twist1 in these cells by RT-PCR.</p><p><b>RESULTS</b>Different levels of Snail, Slug, ZEB1, SIP1 and Twist1 were detected in both LNCaP and LNCaP/HIF1alpha cells, with significant differences only in the expressions of Slug and Twist1 between the two cells. The expression of Slug was increased, but that of Twist1 decreased in the LNCaP/HIF1alpha cells. All the five transcription factors but Twist1 were expressed in both the IF11 and IA8 cells, but only the express- sions of ZEB1 and Slug were increased significantly in the IA8 cells.</p><p><b>CONCLUSION</b>There are different mechanisms underlying transcriptional regulation in different prostate cancer EMT models. Slug may be one of the key transcription factors involved in the HIF1alpha-induced EMT of LNCaP cells, while ZEB1 and Slug may play an important role in repressing the epithelial phenotype of the ARCaP model.</p>

High-dose estrogen inhibits prostate development by down-regulating SHH signaling in new-born SD rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of high-dose estrogen on the development of the prostate in new-born SD rats and its possible relationship with the sonic hedgehog (SHH) signaling pathway.</p><p><b>METHODS</b>One hundred new-born male SD rat pups were assigned to an experimental group, subcutaneously injected with 25 microg Estradiol + 25 microl corn oil vehicle, and a control group, given corn oil alone, on postnatal day (PD) 0, 1, 3 and 5. The animals were sacrificed by decapitation and their prostates removed on PD 1, 6, 10, 20 and 30. The morphological changes of the prostate were observed by HE staining, and SHH signaling related molecules were detected by immunohistochemistry and reverse transcription PCR.</p><p><b>RESULTS</b>High-dose estrogen significantly inhibited the organogenesis of the new-born SD rat prostate and down-regulated the expressions of SHH signaling related molecules.</p><p><b>CONCLUSION</b>High-dose estrogen may restrain the development of the prostate in new-born SD rats via inhibition of SHH signaling.</p>

Transforming growth factor beta upregulates the expression of invasion and metastasis associated proteins in prostate cancer LNCaP cell lines in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the effect of the transforming growth factor beta (TGF-beta) on the expression of invasion and metastasis associated proteins in the prostate cancer LNCaP cell line in vitro.</p><p><b>METHODS</b>The prostate cancer cell line LNCaP was treated with TGF-beta in vitro. Western blotting was used to detect the expression of the "invasion and metastasis" associated proteins E-Cadherin, N-Cadherin and Vimentin.</p><p><b>RESULTS</b>The expression of N-Cadherin and Vimentin of the LNCaP cells treated with TGF-beta for 12 hours was significantly upregulated, but not that of E-Cadherin.</p><p><b>CONCLUSION</b>TGF-beta may induce epithelial-mesenchymal transition (EMT) of LNCaP cells which might be of importance in promoting prostate cancer cells invading to ambient tissues and metastasizing to distant organs.</p>

Comparison of seven screening methods in the diagnosis of bladder cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We compared the validity (evaluated by sensitivity and specificity), reliability (evaluated by reproducibility) and yield (evaluated by predictive value, examining complexity and cost) of individual and combined tests for bladder tumour antigen stat (BTAstat), nuclear matrix protein 22 (NMP22), hyaluronic acid (HA), survivin, CD44v6, vascular endothelial growth factor (VEGF), and voided urine cytology (VUC) in detecting bladder cancer. And at the same time we evaluated the clinical value of these seven detecting methods in the diagnosis of bladder cancer.</p><p><b>METHODS</b>The six markers and VUC were detected in the urine of cancer group (151 patients with bladder cancer) and two control groups (50 patients with benign urological diseases and 50 healthy controls). The sensitivity, specificity, predictive value, reproducibility, examining complexity and checking cost of each marker and combined markers were calculated.</p><p><b>RESULTS</b>There was a significant difference between bladder cancer group and the two control groups. The sensitivity, specificity and positive predictive value were as follows: VUC (36.4%, 100.0%, 100%), BTAstat (76.8%, 87.0%, 89.9%), NMP22 (77.5%, 81.0%, 86.0%), HA (82.8%, 83.0%, 88.0%), survivin (70.2%, 85.0%, 87.6%), CD44v6 (50.3%, 79.0%, 78.4%), and VEGF (68.2%, 93.0%, 93.6%). The highest sensitivities were 91.4% for NMP22 + BTAstat and HA + NMP22, whereas the combined marker with the lowest sensitivity (62.3%) was VUC + CD44v6. The highest specificity was 93.0% for the combined use of VUC + VEGF and HA + CD44v6 had the lowest specificity (73.0%). The most convenient examining method was the detection for BTAstat, the lowest cost was the detection for HA, and the best reproducibility were the detection for BTAstat and VUC.</p><p><b>CONCLUSIONS</b>All the markers have obvious clinical value in diagnosis of bladder cancer. The use of BTAstat + HA or NMP22 + BTAstat are better examining methods in terms of validity, reliability, and yield.</p>

Expression patterns of sonic hedgehog signaling molecules in human fetal prostate development / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate whether the sonic hedgehog signaling pathway is involved in the development of human fetal prostate, and to evaluate the changing staining patterns of its molecules, sonic hedgehog (SHH), patchedl (PTC1), smoothened (SMO), and GLI1, in the human fetal prostate at various gestation stages.</p><p><b>METHODS</b>Fifteen human fetal prostate specimens at various developmental stages (10 - 39 weeks) were included in this study. SHH, PTC1, SMO and GLI1 were detected in all the specimens by immunohistochemical technique. All the slides were observed and assessed under the light microscope.</p><p><b>RESULTS</b>SHH, PTC1, SMO and GLI1 could be detected in human fetal prostate tissues, and their expression formed two surges, the former at week 16, and the latter at week 28. The staining of SHH and SMO was distributed only in the ductal epithelium but not in the stroma. The expression of PTC1 and GLI1 could be found mainly in the epithelium, with minimal staining in the stroma.</p><p><b>CONCLUSION</b>The sonic hedgehog signaling pathway is involved in the development of the human fetal prostate. The high expression of its molecules at early gestation stages might be associated with the induction of prostatic buds, while their abundant expression at later gestation stages might be related to the prostate ductal branching, growth, differentiation and morphogenesis.</p>

Type IV collagenase expression in human prostate carcinoma cell lines with different metastasis potentials / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between type IV collagenase expression and metastatic potential of human prostate carcinoma cells.</p><p><b>METHODS</b>Human prostate carcinoma cell lines (LNCaP and C4-2B) with different metastatic potentials were selected. Using SABC immunocytochemical staining and zymographic analysis, we detected the difference of type IV collagenase expression and activity between the two cell lines.</p><p><b>RESULTS</b>The expression of type IV collagenase (MMP-2, MMP-9) could be detected in both LNCaP and C4-2B, mainly in cytoplasm, much higher in C4-2B than in LNCaP (P < 0.01). The activities of type IV collagenase in the conditioned medium of prostatic carcinoma cell LNCaP was the lowest, and the quantitative estimation values of MMP-2 and MMP-9 were 0.89 +/- 0.02 and 0.86 +/- 0.01, respectively. However, the quantitative estimation values of MMP-2 and MMP-9 in the conditioned medium of C4-2B were 96.32 +/- 4.36 and 33.89 +/- 1.84, respectively. Compared with LNCaP, the activity of type IV collagenase in C4-2B was much higher (P < 0.01).</p><p><b>CONCLUSION</b>Compared with LNCaP, the higher production capability of type IV collagenase in C4-2B may possibly contribute to its invasive and metastatic potentials. The expression of type IV collagenase of human prostate carcinoma cells is closely correlated to the metastatic and invasive potential.</p>

Expression and significance of metastasis-associated proteins in prostate cancer cell lines with different metastatic potentials / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the expression profiles between two metastasis-associated proteins in different prostate cancer cell lines and explore the molecular mechanisms of bone metastatic potentials.</p><p><b>METHODS</b>Expressions of E-cadherin and vimentin in two prostate cancer cell lines (LNCaP and IA8) with different metastatic potentials were detected by Western blotting.</p><p><b>RESULTS</b>There was remarkable difference in the expressions of E-cadherin and vimentin between the highly metastatic cell line and the lowly one. As one of the adhesion associated proteins, E-cadherin was detected with high level of expression in LNCaP cell line, which was well known as low metastatic potential. However, E-cadherin did not expressed in IA8 with high metastatic potential. And as one of the cytoskeleton proteins, vimentin expression was high in IA8, but not in LNCaP.</p><p><b>CONCLUSION</b>There is definitely difference in the metastatic phenotypes (E-cadherin and vimentin) among cell lines with different metastatic potentials. The expressions of E-cadherin and vimentin proteins may play important roles in promoting and inhibiting the metastasis of prostate cancer respectively, and may be considered to be valuable in evaluating the malignant degree, predictable metastasis and prognosis of prostate cancers.</p>

Determination of ofloxacin in human fallopian tube, uterus and serum by high performance liquid chromatography / 药学学报

<p><b>AIM</b>To establish a method for determineation of the concentration of ofloxacin in human fallopian tube, uterus and serum.</p><p><b>METHODS</b>The separation was performed on a Spherisob C18 column (Hypersil, 250 mm x 4.6 mm ID, 5 microns) with a mobile phase of acetonitrile-0.01 moL.L-1 potassium dihydrogen phosphate-0.5 mol.L-1 tetrabutylammonium bromide (9:91:4, pH 2.5). The flow rate was 1.0 mL.min-1 and detection was at 294 nm. The samples were homogenated or ground to powder after freezing with liquid nitrogen. 1% triton-100 and certain volume of ethylacetate-isopropanol (10:1) were added, shaken and centrifuged. Then the entire organic layer was transferred to a tube and vacuum dried. The residue was reconstituted in the mobile phase for HPLC.</p><p><b>RESULTS</b>There was a linear relationship between the peak area ratio and the ofloxacin concentration over the range of 0.2-8.0 micrograms.mL-1. The limits of detection was 40 ng.mL-1. Using this method to determine the ofloxacin concentrations in relevant organs as well as in the plasma of patients of the Department of Gynecology, and achieved satisfactary results.</p><p><b>CONCLUSION</b>The method can be applied to assay the ofloxacin concentration in human tissues. Ofloxacin was well distributed in woman fallopian tube, uterus and serum after single oral administration.</p>