Apalutamide for patients with metastatic castrationsensitive prostate cancer in East Asia: a subgroup analysis of the TITAN trial / 亚洲男科学杂志(英文版)

Ethnicity might be associated with treatment outcomes in advanced prostate cancer. This study aimed to evaluate the efficacy and safety of androgen deprivation therapy (ADT) combined with apalutamide in East Asians with metastatic castration-sensitive prostate cancer (mCSPC). The original phase 3 Targeted Investigational Treatment Analysis of Novel Anti-androgen (TITAN) trial was conducted at 260 sites in 23 countries. This subgroup analysis included patients enrolled in 62 participating centers in China, Japan, and Korea. Radiographic progression-free survival (PFS), time to prostate-specific antigen (PSA) progression, and PSA changes from baseline were compared between groups in the East Asian population. The intent-to-treat East Asian population included 111 and 110 participants in the apalutamide and placebo groups, respectively. The 24-month radiographic PFS rates were 76.1% and 52.3% in the apalutamide and placebo groups, respectively (apalutamide vs placebo: hazard ratio [HR] = 0.506; 95% confidence interval [CI], 0.302-0.849; P = 0.009). Median time to PSA progression was more favorable with apalutamide than placebo (HR = 0.210; 95% CI, 0.124-0.357; P < 0.001). Median maximum percentages of PSA decline from baseline were 99.0% and 73.9% in the apalutamide and placebo groups, respectively. The most common adverse event (AE) was rash in the apalutamide group, with a higher rate than that in the placebo group (37.3% vs 9.1%). The most common grade 3 or 4 AEs were rash (12 [10.9%]) and hypertension (12 [10.9%]) for apalutamide. The efficacy and safety of apalutamide in the East Asian subgroup of the TITAN trial are consistent with the global results.

Efficacy and tolerability of dapoxetine in the treatment of premature ejaculation / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and adverse effects of dapoxetine in the treatment of premature ejaculation.</p><p><b>METHODS</b>We randomly assigned outpatients with premature ejaculation in the proportion of 2:1 to receive 30 mg dapoxetine on demand (n =78) or 50 mg sertraline qd for one month (n = 39). Follow-up was accomplished in 95 cases, 63 in the dapoxetine group and 32 in the sertraline group. We recorded the intravaginal ejaculatory latency time (IELT), clinical global impression of change (CGIC) score, and adverse reactions of the patients and compared them between the two groups.</p><p><b>RESULTS</b>IELT was significantly increased in both the dapoxetine (from [0.87 ± 0.31] to [2.84 ± 0.68] min, P < 0.05) and the sertraline group (from [0.84 ± 0.28] to [2.71 ± 0.92] min, P < 0.05) after medication. Based on the CGIC scores in premature ejaculation, the rate of excellence or effectiveness was 36.5% in the dapoxetine and 37. 5% in the sertraline group, and the rate of improvement was 63.5% in the former and 71.9% in the latter. The incidence rates of dizziness, nausea, headache, and diarrhea were slightly higher (P > 0.05) while those of fatigue, somnolence, and dry mouth significantly higher (P < 0.05) in the sertraline than in the dapoxetine group.</p><p><b>CONCLUSION</b>On-demand oral medication of dapoxetine is effective and well-tolerated for the treatment of premature ejaculation.</p>

Changes of intracellular cholesterol metabolism in neuroendocrine differentiation of prostate cancer and their significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the roles of intracellular cholesterol metabolism in neuroendocrine (NE) differentiation of prostate cancer based on an androgen-independent prostate cancer NE cell model induced by androgen deprivation.</p><p><b>METHODS</b>LNCaP cells were cultured in androgen-depleted medium, and NE phenotypes were identified by observing the changes in cell morphology, molecular markers (SgIII, NSE and CgA) and cell proliferation. The expression and distribution of cholesterol and Sg III were determined by immunofluorescence staining. The expressions of the key genes LDL-R, SREBP-1 and SREBP-2 involved in cholesterol synthesis and uptake were detected by semi-quantitative RT-PCR.</p><p><b>RESULTS</b>The LNCaP cells showed shrinking bodies and extending axons after androgen deprivation, and all the molecular markers, such as Sg III, NSE and CgA, significantly increased in a time-dependent manner, while the cell proliferation was obviously inhibited (P < 0.05). The cholesterol distribution in the LNCaP cells after NE differentiation presented remarkable aggregation at the axon terminals. However, there were no significant differences in the expression of cholesterol between the two types of cells, nor in the changes of the expressions of key genes LDL-R, SREBP-1 and SREBP-2 involved in cholesterol synthesis and uptake (P > 0.05).</p><p><b>CONCLUSION</b>Transient androgen depletion could successfully induce NE differentiation of LNCaP cells, and the intracellular cholesterol could re-distribute into axon terminals to enhance the formation of neurosecretory granules.</p>

Advances in the studies of androgen metabolism and de novo androgen synthesis in castration resistant prostate cancer / 中华男科学杂志

Prostate cancer generally relapses into castration resistant prostate cancer (CRPC) after androgen deprivation therapy, which may be associated with androgen metabolism, particularly de novo androgen synthesis apart from the amplification and mutation of androgen receptor and the activation of its signaling pathways. This article focuses on the advances in the studies of the changes in androgen metabolism and de novo androgen synthesis in CRPC as well as their possible mechanisms and clinical significance.

Establishment of an animal model of prostate cancer metastasis to the lung traced by AsRed2 / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish an animal model of prostate cancer (PCa) metastasis to the lung using PCa PR7 (PCa PC-3 cells stably expressing red fluorescent protein AsRed2) cell lines that can be monitored by in vivo fluorescence imaging technology.</p><p><b>METHODS</b>MTT and Transwell assay were used to compare the abilities of proliferation, migration and invasion of PC-3 and PR7 cells. Twenty BALB/c nude mice were equally randomized to 4 groups to receive tail vein injection of PR7 cell suspension at the concentration of 1 x 107/ml (group A), 2.5 x 107/ml (group B), 5 x 107/ml (group C) and 2.5 x 107/ml followed by the same dose 1 week later (group D). PCa metastasis to the lung was then monitored by in vivo fluorescence imaging technology at the end of 2, 4, 6 and 8 weeks.</p><p><b>RESULTS</b>There were no statistically significant differences between PC-3 and PR7 cells in their abilities of proliferation, migration and invasion (P > 0.05). At the end of 4 weeks, lung metastasis was observed in 40% of the mice in group D, and at the end of 8 weeks, it was detected in 20% in group A, 60% in group B, 100% in group C, and 100% in group D, all confirmed by pathological examination.</p><p><b>CONCLUSION</b>The animal model of PCa metastasis to the lung that can be monitored by in vivo fluorescence imaging technology was established successfully by tail vein injection of PR7 cells carrying red fluorescent protein.</p>

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>

Over-expression of hypoxia-inducible factor-1alpha induces epithelial-mesenchymal transition in LNCaP cells in vitro and in vivo / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the over-expression of hypoxia-inducible factor-1alpha (HIF-1alpha) on the epithelial-mesenchymal transition (EMT) and invasive potency of LNCaP cells in vitro and in vivo.</p><p><b>METHODS</b>We cultured LNCaP cells stably expressing HIF-1alpha (LNCaP/HIF-1alpha) and LNCaP cells, identified the over-expression of HIF-1alpha, determined the proliferation of the two cell lines by MTT assay and the level of PSA in the supernatant of culture medium, and detected the anchorage independent growth by soft-agar colony formation assay. A subcutaneous tumor model was established in nude mice by injecting LNCaP/HIF-1alpha and LNCaP cells followed by observation of the tumor growth. Tumor specimens were obtained for immunohistochemistry.</p><p><b>RESULTS</b>The over-expression of HIF-1alpha was confirmed in the LNCaP/HIF-1alpha cells by immunofluorescence staining and Western blotting. The level of PSA was obviously decreased in LNCaP/HIF-1alpha as compared with that in LNCaP cells. MTT assay identified the increased proliferation of LNCaP/HIF-1alpha cells. The cell colony forming ability of the LNCaP cells was significantly lower than that of the LNCaP/HIF-1alpha cells. The rate of tumorigenesis was increased and its time shortened in the LNCaP/HIF-1alpha group. Immunohistochemistry revealed an up-regulated expression of vimentin and a down-regulated expression of E-cadherin in the tumor specimens.</p><p><b>CONCLUSION</b>The overexpression of HIF-1alpha can up-regulate the expression of vimentin and down-regulate the expression of E-cadherin, which may enhance the invasive potency of LNCaP cells by inducing EMT.</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>

Expression of COX-2 in different human prostate cancer cell lines and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the expression of cyclooxygenase-2 (COX-2) in different prostate cancer (PCa) cell lines and its role in the acquisition of invasive and metastatic potentials of PCa.</p><p><b>METHODS</b>We detected the expressions of COX-2 in prostate cancer cell lines LNCaP, C4-2, IF11, IA8 and PC-3 with different metastatic potentials by Western blotting and RT-PCR, and analyzed their roles in the invasion and metastasis of different PCa cell lines.</p><p><b>RESULTS</b>Western blotting and RT-PCR showed that the expression of the COX-2 protein was high in PC-3, but absent in IF11, IA8, LNCaP and C4-2 (P < 0.05), and it was consistent with the expression of COX-2 mRNA.</p><p><b>CONCLUSION</b>COX-2 expresses differently in PCa cell lines with different metastatic potentials. The overexpression of COX-2 may be associated with the high invasion and metastasis of PC-3, but not with the metastasis of other cell lines.</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>

Expression of Smad4 in prostate cancer LNCaP and ARCaP cell lines with different metastatic potentials and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the expression of Smad4, the core of TGF-beta/Smads signal transduction pathway in different prostate cancer cell lines, and explore their molecular mechanism of bone metastatic potential.</p><p><b>METHODS</b>The Millicell polycarbonate filter coated with matrigel was used to confirm the invasive potency of LNCaP and ARCaP cell lines (IF11 and IA8). The expressions of the Smad4 protein and mRNA in these prostate cancer cells with different metastatic potentials were detected by Western blotting and RT-PCR, respectively.</p><p><b>RESULTS</b>ARCaP cell lines (IF11 and IA8) exhibited a stronger potency of invasion than LNCaP (P < 0.01). The Smad4 protein and mRNA highly expressed in the LNCaP cell line that was well-known with a low metastatic potential, but not in the ARCaP (IF11 or IA8) cells with high metastatic potentials (P < 0.01).</p><p><b>CONCLUSION</b>Smad4 expresses differently in LNCaP and ARCaP cell lines with different metastatic potentials and, as a tumor suppressive gene, its deficient expression may play an important role in the invasion and metastasis of advanced prostate cancer.</p>

Identification of Wnt/beta-catenin signal pathway in human prostate cancer cell lines and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the Wnt/beta-catenin signal pathway in different human prostate cancer cell lines and explore its role in epithelial-mesenchymal transition (EMT) in human prostate cancer.</p><p><b>METHODS</b>We detected the expressions of beta-catenin, t-GSK3beta and p-GSK3beta in several prostate cancer cell lines (LNCaP, C4, C4-2, C4-2B, IF11, IA8, PC-3 and DU145) with different characteristics of epithelial-mesenchymal transition (EMT) by Western blotting.</p><p><b>RESULTS</b>There were remarkable differences in the expressions of beta-catenin and p-GSK3beta among the cell lines, with beta-catenin and p-GSK3beta highly expressed in LNCaP, C4, C4-2, C4-2B, IF11 and IA8, lowly expressed in PC-3 and DU145, but no difference was observed in the expressions of t-GSK3beta in all the cell lines.</p><p><b>CONCLUSION</b>There are differences in the state of the Wnt/beta-catenin signal pathway among the cell lines with different characters of EMT.</p>

Two mutations of the KRT6A gene in Chinese patients with pachyonychia congenita type I / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the gene mutation in a Chinese pedigree and one sporadic case with pachyonychia congenita type I(PC-1), as well as to explore the relationship between the genotype and phenotype.</p><p><b>METHODS</b>The whole coding region of the KRT16 and KRT6A genes were amplified by long-range polymerase chain reaction (PCR). Six patients with PC-1 were studied, five of them were from a pedigree and the other one was sporadic. One unaffected member in the pedigree and 100 unrelated healthy individuals were also studied in order to exclude polymorphism. PCR products were directly sequenced to detect the mutation.</p><p><b>RESULTS</b>No mutations in the KRT16 gene were observed. All patients harbored a mutation in the KRT6A gene. All five patients in the pedigree had a mutation at codon 465 (TAC to CAC) which substitutes tyrosine (Y) by histidine (H). In the sporadic patient, codon 171 (AAC) was mutated to GAC, which changes the asparagines (N) to aspartic acid (D). No such mutations were found in the unaffected member of the pedigree and the 100 unrelated controls. The mutation of Y465H is located at the end of 2B and N171D at the beginning of 1A domain of KRT6A, both are hotspots for pathogenic keratin mutations.</p><p><b>CONCLUSION</b>The mutations Y465H and N171D of the KRT16A gene were detected in the pedigree and the sporadic case respectively. The Y465H mutation was a novel mutation, and the N171D mutation was reported recently.</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>

Expressions of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines and their significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the expressions of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines and explore their relationship with the metastasis of human prostate cancer.</p><p><b>METHODS</b>The expressions and distribution of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines (IF11 and IA8) were detected by Western blot and immunofluorescent staining.</p><p><b>RESULTS</b>The expression of E-cadherin was high in LNCaP, but absent in IF11 and IA8, while beta-catenin was expressed highly in IF11 and LA8, but lowly in LNCaP. Immunofluorescent staining showed that E-cadherin was mainly in the membrane of LNCaP, while beta-catenin both in the membrane of LNCaP and in the nuclei of IF11 and IA8.</p><p><b>CONCLUSION</b>E-cadherin and beta-catenin are differently expressed and distributed in prostate cancer cell lines with different characteristics of epithelial-mesenchymal transition (EMT), and the abnormal activation of the beta-catenin signal pathway may be involved in the EMT of prostate cancer cells.</p>

Identification of TGF-beta/Smads pathway in human prostate cancer cell lines and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the TGF-beta/Smads signal pathway in different human prostate cancer cell lines, and to explore the role of the TGF-beta/Smads pathway in the progression and metastasis of prostate cancer and its possible mechanisms.</p><p><b>METHODS</b>We detected the expressions of the key proteins involved in the TGF-beta/Smads pathway, TbetaR II, Smad2/3, p-Smad2 and Smad4, in prostate cancer cell lines LNCaP, PC-3, DU145, IF11 and IA8 with different metastatic potentials by Western blotting.</p><p><b>RESULTS</b>TbetaR II was expressed highly in PC-3, DU145, IF11 and IA8, but extremely lowly in LNCaP. Smad2/3 was highly expressed in all the cell lines with different intensity, while p-Smad2 only in PC-3 and DU145. The expression of Smad4 was detected in LNCaP, PC-3 and DU145, but not in IF11 and IA8.</p><p><b>CONCLUSION</b>The status of the TGF-beta/Smads pathway differs in the cell lines with different metastatic potentials, only active in PC-3 and DU145. The TGF-beta/Smads pathway may be involved in the invasion and metastasis of prostate cancer through altering the expressions of the key proteins in it.</p>

Relationship between lower urinary tract symptoms and objective measures of benign prostatic hyperplasia: a Chinese survey / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The enlarged prostate leads to obstruction and lower urinary tract symptoms (LUTS), which comprise frequency, urgency, weak stream, straining and nocturia. This study was conducted in a large series of patients to evaluate the relationship between LUTS as stipulated in the International Prostate Symptom Score (IPSS) and the objective parameters related to benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>We enrolled 1295 BPH patients from seven centers. The patients were either at first diagnosis of BPH or had discontinued medical treatment for at least 3 months. Those with several other diseases that may be potential risk factors affecting urinary symptoms were excluded from the study. Age, IPSS, prostate volume, peak flow rate, urine volume and post-voiding residual urine volume were measured. The relationship between IPSS and objective parameters were quantified by means of Spearman correlation coefficients. The differences in these parameters between the groups with mild, moderate or severe symptoms were also evaluated.</p><p><b>RESULTS</b>Statistically significant correlations were found between IPSS and objective parameters by means of Spearman correlation coefficients. When the patients were divided into three groups with different severities of symptoms, there were significant differences in peak flow rate, urine volume, prostate volume, residue urine volume and quality of life, whereas average age and prostate-specific antigen levels were similar. However, there was evident overlap of these parameters between the groups. The same results were found when the irritative or obstructive subscore of IPSS was considered.</p><p><b>CONCLUSIONS</b>The correlation between objective parameters of BPH and LUTS is significant. However, it is hard to predict the severity of symptoms by these parameters.</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>

Over-expression of hypoxia-inducible factor 1 alpha increases angiogenesis of LNCaP cells in vivo / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the influence of hypoxia-inducible factor 1 alpha (HIF-1alpha) on angiogenesis in prostate carcinoma in vivo and to investigate its molecular mechanism.</p><p><b>METHODS</b>LNCaP/HIF-1alpha and LNCaP cells were cultured, the level of PSA in the supernatant of the culture medium detected by ELISA assay before and after the transfection, and the cellular cycle measured by flow cytometry. Nude mouse models of subcutaneous tumor were established with LNCaP/HIF-1alpha and LNCaP cells, the tumor growth observed, and tumor specimens collected for immunohistochemical staining.</p><p><b>RESULTS</b>Compared with the LNCaP cells, LNCaP/HIF-1alpha cells showed an obviously decreased PSA level (t = 8.243, P < 0.05) and enhanced proliferous activity. The tumorigenesis rate increased and the tumorigenesis time advanced in the LNCaP/HIF-1alpha group of the nude mice. Immunohistochemistry displayed higher expressions of VEGF, iNOS and Ang-2 in the LNCaP/HIF-1alpha than in the LNCaP group.</p><p><b>CONCLUSION</b>The over-expression of HIF-1alpha can up-regulate VEGF and iNOS involved in angiogenesis in vivo and contribute to the invasive potency of LNCaP cells. HIF-1alpha may have no influence on Ang-2 either in vitro or in vivo, while the expression of Ang-2 is regulated by other factors in vivo.</p>

Over-expression of HIF-1alpha induces EMT of human prostate cancer cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine whether human prostate cancer cell lines undergo epithelial-mesenchymal transition (EMT) and become more invasive when induced by HIF-1alpha, and to explore the underlying molecular mechanism.</p><p><b>METHODS</b>The cell line LNCaP, appropriate for the HIF-1alpha induction test, was screened out from 4 different EMT-negative prostate cell lines through vimentin gene detection by RT-PCR. The recombinant plasmid pCDNA3. 1(-)/HIF-1alpha was constructed and transfected into LNCaP with the Lipofectamine 2000 system. The control plasmid pCDNA3.1 (-) was transfected by the same method. The positive clone cells were selected by G418 and confirmed by Western blot and immunofluorescence staining. Then a Transwell polycarbonate filter, coated with 100 micol Matrigel at 1:20 dilution in the serum-free medium, was used to analyze the invasive potency. The expression of E-cadherin and vimentin was detected by Western blot.</p><p><b>RESULTS</b>Among the 4 different EMT-negative cell lines, LNCaP was the only one that expressed the vimentin gene but not protein. The expression of HIF1alpha was obviously higher in LNCaP/HIF1alpha than in LNCaP/pCDNA3. 1 (- an LNCaP. The number of the LNCaP/HIF1alpha cells that penetrated through the Transwell polycarbonate filter was significantly larger than that of the LNCaP and LNCaP/pCDNA3. 1(-) cells. Compared with the LNCaP/pCDNA3.1(-) and LNCaP cells, the expression of vimentin was up-regulated, while that of E-cadherin down-regulated, in LNCaP/HIF1alpha.</p><p><b>CONCLUSION</b>The over-expression of HIF-1alpha could induce EMT in the human prostate carcinoma cell line LNCaP and enhance its invasiveness through E-cadherin and vimentin regulation.</p>