Function and regulatory mechanism of androgen receptor variant 7 in castration resistant prostate cancer / 中华泌尿外科杂志

Androgen receptor (AR) plays a key regulatory role in the development of castration resistant prostate cancer (CRPC), and the level of constitutive active variants represented by androgen receptor variant 7 (AR-V7) is increasing during the progress of CRPC, which can be used as a molecular marker of disease progress and prognosis of patients with CRPC. It is an important target to overcome castration resistance and improve the quality of life and survival of patients. In this paper, the function of AR-V7 and its molecular regulation mechanism in CRPC are reviewed. The research shows that the generation of AR-V7 is related to the structural rearrangement of AR gene, gene amplification and the selective splicing of AR gene transcripts, and it is affected by the coordinated regulation of multiple signal pathway molecules such as TGF-β; AR-V7 changes the transport and nuclear localization mechanism of AR protein, and further affects the transcriptional expression of downstream target genes. AR-V7 antagonizes AR activity and blocks the differentiation process driven by AR and androgen, and inhibits the expression of tumor suppressor genes to stimulate the proliferation of tumor cells, thus promoting the progress of Pca. Related targeting studies have revealed AR-V7 targets and CRPC treatment strategies. Currently, they mainly focus on AR-V7 protein degradation, mRNA expression inhibition and N-terminal domain targeting intervention. With the development of in-depth research, the molecular mechanism of AR-V7 in the progress of Pca will be gradually clarified, which will certainly play a greater role in the prevention and treatment of CRPC.

UHRF1/DNMT1-MZF1 axis-modulated intragenic site-specific CpGI methylation confers divergent expression and opposing functions of PRSS3 isoforms in lung cancer

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.

Late Onset Subacute Profound Biotinidase Deficiency Caused by a Novel Homozygous Variant c.466-3T>G in the BTD Gene

Biotinidase defciency (BD) is an autosomal recessive disorder caused by bi-allelic mutation in the BTD gene. Clinical manifestations in BD mainly depends on residual biotinidase enzyme activity, although there are some exceptions. Broadly BD disorders are classifed as profound BD and partial BD. Further profound BD can be early onset, late onset, and sometimes may be asymptomatic. Clinically late-onset profound BD can present with spectrum of manifestations ranging from single organ to multiple organ involvement, typically afecting function of brain, eye, ear, and skin. Here, a frst-born child to consanguineous parents with late-onset profound BD presenting with hyperventilation secondary to lactic acidosis, hypotonia, evolving spasticity, and abnormal neuroimaging fndings caused by novel homozygous variant, c.466-3T>G in the BTD gene is reported.

Exploration of Mechanism of Guben Qingyuan Prescription Combined with Androgen Deprivation Therapy Against Castration-resistant Prostate Cancer from AR/AR-V7 Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the efficacy and mechanism of Guben Qingyuan prescription combined with androgen deprivation therapy （ADT） in the treatment of castration-resistant prostate cancer （CRPC）. Method:A CRPC-bearing mouse model was established. When the tumor volume reached about 100 mm³， 50 CRPC-bearing BALB/c nude mice were randomly divided into the model group， ADT group， and ADT+low-， medium-， high-dose Guben Qingyuan prescription groups， with 10 mice in each group. After grouping， it was ensured that there was no statistically significant difference in tumor volume between groups. The mice in the model group was treated with the same amount of normal saline （10 mL·kg^-1） by gavage， twice a day， while those in the other groups were provided with bicalutamide （5 mg·kg^-1） for intragastric administration， once a day， and then with goserelin （0.36 mg·kg^-1） for intraperitoneal injection on the 10th day. On the basis of ADT， the ones in the ADT+Guben Qingyuan prescription groups further received Guben Qingyuan prescription at the low （2.5 g·kg^-1）， medium （25 g·kg^-1）， and high doses （50 g·kg^-1） by gavage， twice a day. After 25 days of continuous administration， the tumor tissue was harvested for recording the tumor growth and calculating the tumor inhibition rate. The mRNA and protein expression levels of androgen receptor （AR）， androgen receptor splice variant-7 （AR-V7）， and prostate-specific antigen （PSA） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot assay. Result:The tumor inhibition rates of the ADT+low-， medium-， and high-dose Guben Qingyuan prescription groups were 27.95%， 46.71%， and 44.46%， respectively， and the inhibition rates in the ADT+medium- and high-dose Guben Qingyuan prescription groups were significantly increased as compared with that in the ADT group （<italic>P</italic><0.05）. As revealed by comparison with the ADT group， Guben Qingyuan prescription at the medium and high doses significantly down-regulated the mRNA and protein expression levels of AR， AR-V7， and PSA （<italic>P</italic><0.05）. Conclusion:Guben Qingyuan prescription combined with ADT is efficient in controlling the tumor growth in CRPC-bearing mice， which is related to the regulation of AR/AR-V7 signaling pathway.

Conservation of gene architecture and domains amidst sequence divergence in the hsrω lncRNA gene across the Drosophila genus: an in silico analysis

The developmentally active and cell-stress responsive hsrx locus in Drosophila melanogaster carries two exons, one omega intron, one short translatable open reading frame (ORFx), long stretch of unique tandem repeats and an overlapping mir-4951 near its 30 end. It produces multiple long noncoding RNAs (lncRNAs) using two transcription start and four termination sites. Earlier cytogenetic studies revealed functional conservation of hsrx in several Drosophila species. However, sequence analysis in three species showed poor conservation for ORFx, tandem repeat and other regions while the 16 nt at 50 and 60 nt at 30 splice junctions of the omega intron, respectively, were found to be ultra-conserved. The present bioinformatic study using the splice-junction landmarks in D. melanogaster hsrx identified orthologues in publicly available 34 Drosophila species genomes. Each orthologue carries a short ORFx, ultra-conserved splice junctions of omega intron, repeat region, conserved 30 -end located at mir-4951, and syntenic neighbours. Multiple copies of conserved nonamer motifs are seen in the tandem repeat region, despite a high variability in the repeat sequences. Intriguingly, only the omega intron sequences in different species show evolutionary relationships matching the general phylogenetic history in the genus. Search in other known insect genomes did not reveal sequence homology although a locus with similar functional properties is suggested in Chironomus and Ceratitis genera. Amidst the high sequence divergence, the conserved organization of exons, ORFx and omega intron in this gene’s proximal part and tandem repeats in distal part across the Drosophila genus is remarkable and possibly reflects functional importance of higher order structure of hsrx lncRNAs and the small omega peptide.

A novel splice site mutation in OTC gene of a female with ornithine transcarbamylase deficiency and her asymptomatic mosaic father

Ornithine transcarbamylase deficiency is an X-linked disease with a wide range of clinical severity and manifestation age both in males and females. Here, we describe a case which is caused by a novel c.78-1G[A splice site mutation, which on mRNA level leads to a 1-bp deletion and a frameshift (c.78delG (p.C27Vfs*11)) in OTC exon 2 in a young girl. The same mutation has been detected in a mosaic state in her asymptomatic father.

Hereditary spherocytosis due to a novel c. 5798+ 1G>A variant of the SPTB gene / 中华医学遗传学杂志

Objective@#To explore the genetic basis of a pedigree affected with hereditary spherocytosis.@*Methods@#Peripheral blood samples were collected from 17 members of the pedigree. Genomic DNA of the proband was subjected to next generation sequencing. Candidate variant was validated by co-segregation analysis. pCAS2c.5798+ 1G and pCAS2c.5798+ 1A plasmids were constructed by homologous recombination and transfected into 293T cells. Reverse transcription PCR, TA cloning and Sanger sequencing were used to analyze the effect of candidate variant on splicing. Meanwhile, peripheral blood RNAs were extracted to analyze the effect of candidate variant on splicing in vivo.@*Results@#The proband was found to carry a c. 5798+ 1G>A variant of the SPTB gene. The variant has co-segregated with the phenotype in the pedigree. In vitro and in vivo splicing experiments confirmed that the mutation has significantly affected the splicing, resulting in shift of reading frame and produced a premature termination codon.@*Conclusion@#The novel c. 5798+ 1G>A variant of the SPTB gene probably underlies the pathogenesis of hereditary spherocytosis in this pedigree.

Novel splicing mutation of SDHB gene induced paraganglioma near inferior vena cava: A case report / 中华内分泌代谢杂志

Pheochromocytoma/paraganglioma(PPGL) was a kind of neuroendocrine tumor that derived from chromaffin tissue, which seems to be an important etiology of secondary hypertension. With the development of molecular detection technology, at least 17 kinds of pathogenic genes of PPGL has been discovered, which is related to 35%-40% PPGL, and about 40% malignant PPGL is associated with SDHB gene mutation. In this study, we reported a case with a novel splicing mutation of SDHB gene induced paraganglioma.

Impact of taxanes on androgen receptor signaling / 亚洲男科学杂志(英文版)

The development and progression of metastatic castration-resistant prostate cancer is the major challenge in the treatment of advanced prostate cancer. The androgen receptor signaling pathway remains active in metastatic castration-resistant prostate cancer. Docetaxel and cabazitaxel are the first- and second-line chemotherapy, respectively, for patients with metastatic castration-resistant prostate cancer. These two taxanes, in general, function by (i) inhibiting mitosis and inducing apoptosis and (ii) preventing microtubule-dependent cargo trafficking. In prostate cancer, taxanes have been reported to inhibit the nuclear translocation and activity of the androgen receptor. However, whether this is attainable or not clinically remains controversial. In this review, we will provide a comprehensive view of the effects of taxanes on androgen receptor signaling in prostate cancer.

Effect of ovarian granulosa cell androgen receptor splice variant on endocrine hormone levels in patients with polycystic ovary syndrome / 上海交通大学学报（医学版）

Objective • To investigate the effects of ovarian granulosa cell androgen receptor splice variant insertion isoform (AR-ins) on the levels of sex hormones and insulin in patients with polycystic ovary syndrome (PCOS). Methods • Seventy-nine control patients (control group) of normal ovulation and one hundred and fifty-four PCOS patients (PCOS group) who received in vitro fertilization embryo transfer in the Assisted Reproductive Department of the International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine from Jun. 2017 to Jun. 2018 were recruited. The sociodemographic information, serum basic hormones and insulin data were collected according to medical history record, and the hormone levels were measured by chemiluminescence. The ovarian granulosa cells and follicular fluids of all the patients were collected. Nested PCR was used to identify the androgen receptor splice variant (AR SV) in the ovarian granulosa cells from PCOS patients, and then they were divided into two groups, i.e., PCOS Wt group (n=39) and PCOS AR-ins group (n=115). The levels of sex hormones and insulin in follicular fluids of the three groups were detected by chemiluminescence. The free androgen index (FAI) was calculated. The sociodemographic information, the levels of sex hormones and insulin in serum and follicular fluid of the three groups were compared by One-way ANOVA, Welch's ANOVA, Kruskal-Wallis H test and chi-square test. Results • Compared with the control group, the levels of serum sex hormone-binding globulin (SHBG) were significantly lower in the PCOS AR-ins group (P=0.000), and the levels of serum total testosterone (TT) (P=0.000), insulin (P=0.001) and FAI (P=0.000) were significantly higher in the PCOS AR-ins group. Compared with the control group and the PCOS Wt group, the differences of SHBG, TT, insulin levels and FAI in the follicular fluid in the PCOS AR-ins group were statistically significant (all P<0.05). Conclusion • PCOS patients with AR-ins have high androgen and insulin levels in serum, and this effect is more pronounced in the ovarian local microenvironment.

Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused by mutations in the DMD gene. The aim of this study is to identify pathogenic DMD variants in probands and reduce the risk of recurrence of the disease in affected families. Variations in 100 unrelated DMD/BMD patients were detected by multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing (NGS). Pathogenic variants in DMD were successfully identified in all cases, and 11 of them were novel. The most common mutations were intragenic deletions (69%), with two hotspots located in the 5′ end (exons 2–19) and the central of the DMD gene (exons 45–55), while point mutations were observed in 22% patients. Further, c.1149+1G>A and c.1150-2A>G were confirmed by hybrid minigene splicing assay (HMSA). This two splice site mutations would lead to two aberrant DMD isoforms which give rise to severely truncated protein. Therefore, the clinical use of MLPA, NGS, and HMSA is an effective strategy to identify variants. Importantly, eight embryos were terminated pregnancies according to prenatal diagnosis and a healthy boy was successfully delivered by preimplantation genetic diagnosis (PGD). Early and accurate genetic diagnosis is essential for prenatal diagnosis/PGD to reduce the risk of recurrence of DMD in affected families.

ARV-7: A biomarker for the treatment of metastatic castration-resistant prostate cancer / 中华男科学杂志

As more and more patients with metastatic prostate cancer develop resistance to androgen-deprivation therapy (ADT) and consequently castration-resistant prostate cancer (CRPC), reasonable selection of therapies is becoming increasingly important for the prediction of the therapeutic results. Many studies show that androgen receptor splice variant 7 (AR-V7) is involved in the development and progression of CRPC and that the expression of AR-V7, absolutely higher in CRPC than in hormone-nave prostate cancer, plays a significant role in the mechanisms of resistance to abiraterone, enzalutamide and taxane chemotherapies. Further more, some clinical trials have revealed that the AR-V7 level may indicate the prognosis of different therapeutic options: AR-V7 negative in circulating tumor cells suggesting the effectiveness of a new hormonal therapy and taxane chemotherapy while AR-V7 positive indicating the poor result of a new hormonal therapy. These findings show that AR-V7 could be a biomarker for therapeutic options and the prognostic evaluation of CRPC.

Impact of taxanes on androgen receptor signaling / 亚洲男科学杂志(英文版)

The development and progression of metastatic castration-resistant prostate cancer is the major challenge in the treatment of advanced prostate cancer. The androgen receptor signaling pathway remains active in metastatic castration-resistant prostate cancer. Docetaxel and cabazitaxel are the first- and second-line chemotherapy, respectively, for patients with metastatic castration-resistant prostate cancer. These two taxanes, in general, function by (i) inhibiting mitosis and inducing apoptosis and (ii) preventing microtubule-dependent cargo trafficking. In prostate cancer, taxanes have been reported to inhibit the nuclear translocation and activity of the androgen receptor. However, whether this is attainable or not clinically remains controversial. In this review, we will provide a comprehensive view of the effects of taxanes on androgen receptor signaling in prostate cancer.

Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused by mutations in the DMD gene. The aim of this study is to identify pathogenic DMD variants in probands and reduce the risk of recurrence of the disease in affected families. Variations in 100 unrelated DMD/BMD patients were detected by multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing (NGS). Pathogenic variants in DMD were successfully identified in all cases, and 11 of them were novel. The most common mutations were intragenic deletions (69%), with two hotspots located in the 5' end (exons 2-19) and the central of the DMD gene (exons 45-55), while point mutations were observed in 22% patients. Further, c.1149+1G>A and c.1150-2A>G were confirmed by hybrid minigene splicing assay (HMSA). This two splice site mutations would lead to two aberrant DMD isoforms which give rise to severely truncated protein. Therefore, the clinical use of MLPA, NGS, and HMSA is an effective strategy to identify variants. Importantly, eight embryos were terminated pregnancies according to prenatal diagnosis and a healthy boy was successfully delivered by preimplantation genetic diagnosis (PGD). Early and accurate genetic diagnosis is essential for prenatal diagnosis/PGD to reduce the risk of recurrence of DMD in affected families.

Clinical profile in arrhythmogenic cardiomyopathy and a recessive plakophilin-2 gene mutation

Objective Arrhythmogenic cardiomyopathy (ACM) is not an uncommon cause of cardiac morbidity in Kashmir valley. This study was designed to document various clinical features and to sequence exons 11 and 12 of plakophilin 2 (PKP2) gene in these patients. Methods ACM patients who attended cardiology outpatient department of our institute from January 2014 to April 2015 were included in the study. Their records were reviewed. Controls were randomly selected, who had no history or family history of cardiac illness and had a normal cardiac examination. A blood sample was also taken from both the groups for sequencing of exon 11 and 12 of PKP2 gene. ACM patients were followed up until July 2016. Results Eleven ACM patients and seven controls were included in the study. Most common mode of presentation was ventricular tachycardia (VT). Two patients had left ventricular (LV) systolic dysfunction. One patient had a splice site mutation in exon 12 of PKP2 gene and one patient died during follow-up. One of the controls had an intronic variation that has no pathogenic significance vis-à-vis ACM. Conclusion Our study describes various clinical parameters in ACM patients and a recessive plakophilin 2 mutation after a limited PKP2 gene sequencing.

Genetic analysis of a family with Usher syndrome / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To analyze the clinical features of a Usher syndrome family and explore the pathogenic gene of the disease. <p>METHODS: A Chinese family with Usher syndrome was involved in our study. After informed consent, careful clinical examinations were taken and 4mL blood were obtained. The whole genome DNA was extracted and target-captured next generation sequencing of the proband was performed to identify suspected mutations. We used Sanger sequencing to verify the detected mutations in all the members of the family, as well as in 100 normal controls. <p>RESULTS: In addition to typical retinitis pigmentosa, the patients suffered from mild to moderate sensorineural deafness. Sequencing results revealed compound heterozygous mutations(c.2310_2311insA and c.8559-2A>G)of <i>USH2A</i> gene in the patients, and either of the mutations was found in normal relatives that had consanguinity with the patients. Both of the mutations were not found in other members of the family and normal individuals. <p>CONCLUSION: <i>USH2A</i> is the pathogenic gene of the disease in this family. The mutation c.8559-2A>G(IVS42)is a previously reported mutation, while the mutation c.2310_2311 insA(p.E771Rfs*8)is a novel mutation. The study has expanded the mutation spectrum of <i>USH2A</i> gene resulting in Usher syndrome.

The predictive role of the expression ratio of androgen receptor splice variant-7 and androgen receptor in the prognosis of prostate cancer treated by castration therapy / 中华泌尿外科杂志

Objective To evaluate the prognostic value of the ratio of AR and AR-V7 expression in prostate cancer treated by castration therapy.Methods Immunohistochemical staining was performed in biopsy specimen of 136 prostate cancer patients received hormone therapy in Tongji Hospital of Huazhong University of Science and Technology from January 2010 to December 2015.Expression was determined using modified H score method.Patients aged from 53-96,the median age was 71,median tPSA value at diagnosed was 110.00 ng/ml(2.61-4 003.4 ng/ml),median fPSA value was 14.62 ng/ml(0.12-640.19 ng/ml),median PSA density was 1.15 ng/(ml · cm3) [0.02-62.63 ng/(ml · cm3)].Among these,88 (64.7％)patients were diagnosed Gleason score≥8,39(28.7％) patients with Gleason score 7,while 7 (6.6％) patients Gleason score ＜7.There were 54(39.7％) patients diagnosed T4 stage,57(41.9％)patients T3 stage and 25 (18.4％) patients Tx stage;62 (45.6％) patients were diagnosed N 1 stage,46 (33.8％) N0 stage and 28(20.6％) patients Nx stage;97(71.3％) patients were diagnosed M1 stage,30(22.1％) M0 stage,9 (6.6％) patients Mx stage.Cause-specific Cox regression and Kaplan-Meier Analysis were used to analyze the prognosis risk.Results The median follow-up time was 44 months,ranged 15-71 months.During the surveillance,the disease progression-free survival time ranged from 5-59 month,median 19 months.The overall survival time ranged from 12-61 months,median 31 months.Among these,79(58.1％) patients were AR positive and 26(19.1％) patients were AR-V7 positive,while AR and AR-V7 expression had no significant correlation (Spearman-test r =0.042,P =0.629).The AR-V7 positive patients showed significantly lower CRPC progression free survival (10.8 months vs.25.0 months,P ＜ 0.001) and much lower overall survival (20.3 months vs.42.8 months,P ＜ 0.001).The high AR-V7/AR expression ratio group showed significantly lower CRPC progression free survival (12.0 months vs.24.8 months,P ＜ 0.001) and much lower overall survival (22.5 months vs.42.8 months,P ＜ 0.001).In univariate Cox regression analyses,Gleason score at diagnosis,T stage,tPSA,PSA density and high AR-V7/AR expression ratio could predict the prognosis of hormonal therapy.While in multivariate Cox regression analyses,T stage(HR =2.597,95％ CI 1.351-4.995,P =0.004) and high AR-V7/AR expression ratio(HR =5.788,95％ CI 2.530-13.242,P ＜ 0.001) could effectively and independently predict the prognosis of hormonal therapy.Conclusion High AR-V7/AR HS ratio is the independent predictor of the prognosis of prostate cancer hormone therapy.AR-V7 positive and high AR-V7/AR HS ratio patients may have shorter PFS and overall survival time than AR-V7 negative and low AR-V7/AR HS ratio patients.

The study of CUDC-101 inhibits the expression of AR-V7 in castration-resistant prostate cancer cells / 中华泌尿外科杂志

Objective To solve the problem of castration resistant prostate cancer (castrationresistant prostate cancer,CRPC) the problem of drug resistance by studing the expression of the CUDC-101 inhibitor of castration resistant prostate cancer androgen receptor splice variant 7.Methods In this study,the expression of AR-V7 protein in prostate cancer cell lines PC-3,VCaP,22Rv1,LNCap was detected by imnmunoblotting between April 2015 and April 2016,and the highest expression level of cell lines was selected follow-up experiments.Through the cell proliferation and activity experiments,the epigenetic inhibitors:histone deacetylase inhibitor CUDC-101,histone methylation inhibitor DZNeP,DNA methylation inhibitor gemcitabine,histone acetyltransferase inhibitor MG149 to select an inhibitor that reduces the expression of AR-V7 protein in CRPC cells.22Rv1 cells were treated with 30 nmol and 300 nmol of CUDC101 and 1,10 and 20 μmol of Enzalutamide (MDV3100),respectively,and their inhibitory effects on the growth of 22Rv1 cells were examined.Results The results of immunoblotting showed that AR-V7 protein was only expressed in CRPC cell line 22Rv1 and negative in non-CRPC cell line.The expression of AR-V7 in 22Rv1 cells treated with CUDC-101 was significantly lower than that of negative control.While other inhibitors had no effect on the expression of AR-V7.In the cell proliferation and activity assay,the inhibitory rates of 30 nmol CUDC-101 and 1,10 and 20 μmol MDV3100 were 10％,35％ and 45％,respectively,higher than that of MDV3100 alone.28％ and 42％,the difference was statistically significant (P ＜ 0.05).The inhibitory rates of 300 nmol CUDC-101 and MDV3100 were 30％,60％ and 65％,respectively,which were significantly higher than those of MDV3100 alone (P ＜ 0.05).Conclusions CUDC-101 can inhibit the castration resistant prostate cancer androgen receptor splice variant 7 expression,and solved the resistance problem of CRPC.

Effect of AR-V7 on endocrine therapy resistance of prostate cancer / 中国病理生理杂志

AIM:To explore the effect of androgen receptor splice variant 7 ( AR-V7 ) on endocrine therapy resistance of prostate cancer cells and the resistance mechanisms .METHODS:Four prostate cancer cell lines were trans-fected with AR-V7 siRNA ( siAR-V7) using Lipofectamine 2000 kit, and the transfected cells were named as PC 3-siAR-V7, DU145-siAR-V7, LNCaP-siAR-V7 and ArCaP-siAR-V7 cells.The prostate cancer cells transfected with negative con-trol ( NC) siRNA served as negative controls .The expression of AR-V7 at mRNA and protein levels was detected by real-time PCR and Western blot , respectively .The cell viability and cell migration rate were measured by MTT assay and Tran-swell method, respectively.The promoter activity of AR and the protein levels of targeted prostate-specific antigen (PSA) and FK506-binding protein 5 (FKBP5) were monitored by luciferase reporter gene assay and Western blot , respectively. The bicalutamide-resistant cell line, LNCaP-DR, was constructed, and the subcellular localization of AR and AR-V7 pro-teins in LNCaP, LNCaP-siAR-V7 and LNCaP-DR cells was observed by the method of immunofluorescence .The protein in-teraction of AR-V7 and heat shock protein 90 ( HSP90) was determined by co-immunoprecipitation .RESULTS:The mR-NA level of AR-V7 in the 4 prostate cancer cell lines was significantly higher than that in normal prostate epithelial cell line RWPE-1 (P<0.05).The AR-V7 level, cell viability and cell migration rate in the cells transfected with siAR-V7 were notablely lowered compared with the NC siRNA-transfected cells (P<0.05).The cell viability was gradually decreased following with the increase in bicalutamide dose , and down-regulation of AR-V7 expression significantly enhanced the sensi-tivity to bicalutamide (P<0.05).Down-regulation of AR-V7 expression significantly inhibited AR promoter activity and reduced the protein levels of PSA and FKBP5 (P<0.05).The results of immunofluorescence observation showed that most AR and AR-V7 were mainly located in the nucleus , a few AR was located in the cytoplasm , and down-regulation of AR-V7 expression inhibited AR nuclear transport .AR was entirely located in the nucleus and the protein levels of AR-V7 was sig-nificantly increased in the bicalutamide-resistant cells .The interaction of endogenous AR-V7 with HSP90 was found in the prostate cancer cells .CONCLUSION: High AR-V7 level is found in the prostate cancer cells , and down-regulation of AR-V7 expression inhibits the cell viability and migration .High AR-V7 level is related to the bicalutamide resistance .The possible mechanism is that AR nuclear transport is mediated by the interaction of AR -V7 with HSP90 to activate AR signal pathway and regulate targeted gene transcriptional activity , thus resulting in drug resistance .

Effect of AR-V7 on endocrine therapy resistance of prostate cancer / 中国病理生理杂志

AIM:To explore the effect of androgen receptor splice variant 7 ( AR-V7 ) on endocrine therapy resistance of prostate cancer cells and the resistance mechanisms .METHODS:Four prostate cancer cell lines were trans-fected with AR-V7 siRNA ( siAR-V7) using Lipofectamine 2000 kit, and the transfected cells were named as PC 3-siAR-V7, DU145-siAR-V7, LNCaP-siAR-V7 and ArCaP-siAR-V7 cells.The prostate cancer cells transfected with negative con-trol ( NC) siRNA served as negative controls .The expression of AR-V7 at mRNA and protein levels was detected by real-time PCR and Western blot , respectively .The cell viability and cell migration rate were measured by MTT assay and Tran-swell method, respectively.The promoter activity of AR and the protein levels of targeted prostate-specific antigen (PSA) and FK506-binding protein 5 (FKBP5) were monitored by luciferase reporter gene assay and Western blot , respectively. The bicalutamide-resistant cell line, LNCaP-DR, was constructed, and the subcellular localization of AR and AR-V7 pro-teins in LNCaP, LNCaP-siAR-V7 and LNCaP-DR cells was observed by the method of immunofluorescence .The protein in-teraction of AR-V7 and heat shock protein 90 ( HSP90) was determined by co-immunoprecipitation .RESULTS:The mR-NA level of AR-V7 in the 4 prostate cancer cell lines was significantly higher than that in normal prostate epithelial cell line RWPE-1 (P<0.05).The AR-V7 level, cell viability and cell migration rate in the cells transfected with siAR-V7 were notablely lowered compared with the NC siRNA-transfected cells (P<0.05).The cell viability was gradually decreased following with the increase in bicalutamide dose , and down-regulation of AR-V7 expression significantly enhanced the sensi-tivity to bicalutamide (P<0.05).Down-regulation of AR-V7 expression significantly inhibited AR promoter activity and reduced the protein levels of PSA and FKBP5 (P<0.05).The results of immunofluorescence observation showed that most AR and AR-V7 were mainly located in the nucleus , a few AR was located in the cytoplasm , and down-regulation of AR-V7 expression inhibited AR nuclear transport .AR was entirely located in the nucleus and the protein levels of AR-V7 was sig-nificantly increased in the bicalutamide-resistant cells .The interaction of endogenous AR-V7 with HSP90 was found in the prostate cancer cells .CONCLUSION: High AR-V7 level is found in the prostate cancer cells , and down-regulation of AR-V7 expression inhibits the cell viability and migration .High AR-V7 level is related to the bicalutamide resistance .The possible mechanism is that AR nuclear transport is mediated by the interaction of AR -V7 with HSP90 to activate AR signal pathway and regulate targeted gene transcriptional activity , thus resulting in drug resistance .