A Novel KIF4A-Related Model for Predicting Immunotherapy Response and Prognosis in Kidney Renal Clear Cell Carcinoma.

BACKGROUND: The efficacy of chemotherapy in treating Kidney Renal Clear Cell Carcinoma (KIRC) is limited, whereas immunotherapy has shown some promising clinical outcomes. In this context, KIF4A is considered a potential therapeutic target for various cancers. Therefore, identifying the mechanism of KIF4A that can predict the prognosis and immunotherapy response of KIRC would be of significant importance. METHODS: Based on the TCGA Pan-Cancer dataset, the prognostic significance of the KIF4A expression across 33 cancer types was analyzed by univariate Cox algorithm. Furthermore, overlapping differentially expressed genes (DEGs1) between the KIF4A high- and lowexpression groups and DEGs2 between the KIRC and normal groups were also analyzed. Machine learning and Cox regression algorithms were performed to obtain biomarkers and construct a prognostic model. Finally, the role of KIF4A in KIRC was analyzed using quantitative real-time PCR, transwell assay, and EdU experiment. RESULTS: Our analysis revealed that KIF4A was significant for the prognosis of 13 cancer types. The highest correlation with KIF4A was found for KICH among the tumour mutation burden (TMB) indicators. Subsequently, a prognostic model developed with UBE2C, OTX1, PPP2R2C, and RFLNA was obtained and verified with the Renal Cell Cancer-EU/FR dataset. There was a positive correlation between risk score and immunotherapy. Furthermore, the experiment results indicated that KIF4A expression was considerably increased in the KIRC group. Besides, the proliferation, migration, and invasion abilities of KIRC tumor cells were significantly weakened after KIF4A was knocked out. CONCLUSION: We identified four KIF4A-related biomarkers that hold potential for prognostic assessment in KIRC. Specifically, early implementation of immunotherapy targeting these biomarkers may yield improved outcomes for patients with KIRC.

SAMD5 mRNA was overexpressed in prostate cancer and can predict biochemical recurrence after radical prostatectomy.

PURPOSE: To identify a novel biomarker that can predict biochemical recurrence (BCR) after radical prostatectomy. METHODS: The gene expression profile of SAMD5 in prostate cancer was explored based on the oncomine database and The Cancer Genomic Atlas (TCGA). The follow-up information and clinical pathologic variables were extracted from the following cohort study: TCGA_prostate carcinoma. And then, survival analysis was conducted using the Kaplan-Meier plot and Cox's proportional hazard regression model. Furthermore, another independent cohort study: Taylor prostate, was also acquired to validate the predictive effect of SAMD5 on BCR. In addition, the expression profile of SAMD5 in other cancer types was investigated using TCGA dataset. RESULTS: SAMD5 mRNA was shown to be up-regulated in multiple microarray datasets of prostate cancer with the strict statistic criteria: p < 0.01 and fold change ≥ 2. In TCGA_PCa cohort study, high expression of SAMD5 was a risk factor for patients on post-operative BCR (HR 2.181, 95%CI 1.199-3.966, p = 0.011) and this predictive ability was independent of Gleason score and pathologic T stage (HR 2.018, 95%CI 1.102-3.698, p = 0.023). In another validating cohort study, the statistic trend was similar, and the pooled analysis by combining the two cohort study further confirmed its prognostic effect. CONCLUSION: SAMD5 mRNA was overexpressed in prostate cancer and had powerful prognostic ability on predicting post-operative BCR, independent of Gleason score and pathologic T stage. Its high expression was associated with poor prognosis after RP.

Analysis of immune status after iodine-125 permanent brachytherapy in prostate cancer.

BACKGROUND: Permanent prostate brachytherapy (PPB) is an effective treatment choice for low and intermediate risk prostate cancer (PCa). However, the impact of PPB on tumor immune status is still poorly understood. This study aimed to assess the immune status in PCa patients before and at different time points after PPB (1, 3, 6, and 12 months). METHODS: Blood was collected from 32 patients with low and intermediate risk PCa and 12 healthy volunteers. The frequency of immunocompetent cells was identified by flow cytometry. The concentration of immunoglobulins and complements was detected by ELISA. RESULTS: Various immunocompetent cells were dysregulated in PCa patients compared with healthy volunteers. Peripheral serum prostate-specific antigen (PSA) decreased rapidly at the first month after PPB treatment, and the peripheral serum PSA became very low at 6 months after PPB treatment. CD3+ T cells, CD4+ T cells, CD3-CD16+/56+ natural killer (NK) cells were increased significantly at certain time points after PPB. Although the percentage of the CD8+ T cells did not change markedly, the ratio of CD4/CD8 increased significantly at 3 months after PPB (P=0.0196). There was no influence of PPB on B cells number, but the concentration of immunoglobulins IgM, IgG, and IgA, and complements C3 and C4 in patients increased at some time points after PPB. CONCLUSION: The immunocompetent cells are dysregulated in PCa patients. PPB treatment could effectively kill tumor cells and then stimulate cellular immunity and humoral immunity in PCa patients.

[Apoptosis induced by transferring cytosine deaminase suicide gene into kinds of cancer cells with novel polyamidoamine dendrimers].

This study demonstrates efficacy of a novel polyamidoamine dendrimers(PAMAM dendrimers) with pentaerythritol derivatives as the core(G5 PD dendrimer) in deliver of the cytosine deaminase(CD) gene and EGFP gene fusion plasmid into different tumor cell lines to induce apoptosis. The physical and chemical properties of G5 PD dendrimers in terms of DNA complexation, particulate properties and transfection efficiencies were investigated and compared with commercial gene vectors PEI 25 k Da. The optimum ratio of G5 PD dendrimer complexed with plasmid DNA was 0.2∶1, and the particle size of the complex was(100 ± 5) nm. Compared with the commercial gene carriers PEI, G5 PD dendrimer exhibited a higher transfection efficiency at the weight ratio of 1∶1 in three different cell lines, given the fact that PEI are different from PAMAM dendrimers in terms of molecular structure. Furthermore, the cytotoxicity assays of the cell lines transfected with G5 PD dendrimer/p CD-EGFP-N1 followed by exposure to various concentrations of 5-fluorocytosine(5-FC) also showed that the transfected cell lines could generate a very low amount of 5-FC to 5-fluorouracil(5-FU) in a short period of time, which indicating the high expression level of CD gene in the cell line. These results indicate that the CD/5FC system of G5 PD dendrimer has an excellent efficacy in gene delivery.

Formation of vasculogenic mimicry in bone metastasis of prostate cancer: correlation with cell apoptosis and senescence regulation pathways.

Vasculogenic mimicry (VM) has been found in prostate cancer (PCa) as an independent marker of poor prognosis. To investigate the correlation between VM and bone metastasis in PCa, a total of 80 cases were analyzed by CD31 and PAS dual-staining as well as the follow-up data. All cases were divided into two groups: VM-positive and VM-negative (VM-pos/VM-neg). Immunohistochemical staining for investigating the expression of Casepase-3, Bcl-2/Bax, and SA-ß-gal was performed. 28 of the 80 PCa cases exhibited VM structure (35.0%). The incidence of bone metastasis in the VM-pos and VM-neg was 67.9% (19/28) and 38.5% (20/52), respectively. The positive rate of Casepase-3 and Bcl-2 expression was significantly different of the two groups (Caspases-3: VM-pos 71.4%, 20/28 vs VM-neg 42.3%, 22/52; Bcl-2: VM-pos 35.7%, 10/28 vs VM-neg 65.4%, 34/52). Bcl-2/Bax ratio of the VM-pos (0.71±0.22) was lower than that of the VM-pos (0.89±0.13). In addition, a higher frequency of SA-ß-gal was detected in VM-pos (64.29±86.42) than in VM-neg (25.37±72.21). Taken together, our findings demonstrate that PCa with VM has the tendency to develop bone metastasis. Activations of cell apoptosis and senescence regulation pathways may play important roles in the formation process of VM structure.

[Clinical analysis of percutaneous nephrolithotomy for staghorn calculi with different stone branch number].

OBJECTIVE: To investigate the impact of staghorn stone branch number on outcomes of percutaneous nephrolithotomy (PNL). METHODS: From January 2009 to January 2013, the 371 patients with staghorn stones who were referred to our hospital for PNL were considered for this study. All calculi were showed with CT 3-dimentional reconstruction (3-DR) imaging. The computerized database of the patients had been reviewed. Our exclusion criterion was patients with congenital renal anomalies, such as horse-shoe and ectopic kidneys. And borderline stones that branched to one major calyx only were also not included. From 3-DR images, the number of stone branching into minor renal calices was recorded. We made "3" as the branch breakdown between groups. And the patients were divided into four groups. The number of percutaneous tract, operative time, staged PNL, intra-operative blood loss, complications, stone clearance rate, and postoperative hospital day were compared. RESULTS: The 371 patients (386 renal units) underwent PNL successfully, included 144 single-tract PNL, 242 multi-tract PNL, 97 staged PNL. The average operative time was (100 ± 50) minutes; the average intra-operative blood loss was (83 ± 67) ml. The stone clearance rate were 61.7% (3 days) and 79.5% (3 months). The postoperative hospital stay was (6.9 ± 3.4) days. A significantly higher ratio of multi-tract (χ(2) = 212.220, P < 0.01) and staged PNL (χ(2) = 49.679, P < 0.01), longer operative time (F = 4.652, P < 0.01) and postoperative hospital day (F = 2.067, P = 0.043) and lower rate of stone clearance (χ(2) = 10.691 and 47.369, P < 0.05) were found in PNL for calculi with stone branch number ≥ 5. There was no statistically meaningful difference among the 4 groups based on Clavien complication system (P = 0.460). CONCLUSION: The possibility of multi-tract and staged PNL, lower rate of stone clearance and longer postoperative hospital day increase for staghorn calculi with stone branch number more than 5.

[Expression of PIM-1 in prostate cancer tissue and its relationship with PSA recurrence].

OBJECTIVE: To explore the expression of the PIM-1 protein in prostate cancer tissue and its relationship with PSA recurrence. METHODS: We used the immunohistochemical SP method to detect the expression of the PIM-1 protein in the prostate tissues of 68 cases of prostate cancer (PCa) and 37 cases of benign prostatic hyperplasia (BPH). RESULTS: The positive rate of the PIM-1 protein expression was 67.65% (46/68) in the PCa tissue, significantly higher than 40.54% (15/37) in the BPH tissue (P<0.05). Its positive rates in PCa Gleason scores 6, 7 and 8-10 were 33.33% (7/21), 77.5% (21/28) and 94.74% (18/19), respectively, with significant between-group differences (P<0.05), and those in stages I , II, III and IV of PCa were 47.62%, 53.85%, 73.33% and 94.74%, respectively. Kaplan-Meier analysis of the results of a 36-month follow-up showed the ratios of PIM-1 expression to PSA recurrence and non-recurrence were 10/22 (45.45%) and 36/46 (78.26%), respectively, with statistically significant differences (P<0.05). CONCLUSION: PIM-1 protein expression in PCa tissue is closely related to the Gleason score and clinical stage of PCa and PSA recurrence, which suggests that the PIM-1 gene plays an important role in PCa evolution and progression, and may be an indicator for the prognosis of PCa.

[Correlation of histological prostatitis with PSA, prostate volume, PSAD, IPSS, Qmax and PVR in BPH patients].

OBJECTIVE: To explore the correlation of histologically proven prostatitis with the level of prostate specific antigen (PSA), prostate volume, PSA density (PSAD), international prostate symptom score (IPSS), maximum flow rate (Qmax) and post-void residual volume (PVR) in men with symptoms of benign prostate hyperplasia (BPH). METHODS: Totally 673 patients surgically treated for BPH were divided into Groups A and B in accordance with histological findings, the former including those with histological prostatitis, and the latter without it. Comparisons were made between the two groups in the PSA level, prostate volume, PSAD, IPSS, Qmax and PVR. RESULTS: The PSA level, prostate volume, IPSS and PVR were significantly higher in Group A ([5.64 +/- 2.48] microg/L, [43.66 +/- 13.11] ml, 24.72 +/- 5.39 and [124.90 +/- 49.80] ml) than in B ([4.97 +/- 1.99] microg/L, [40.41 +/- 11.44] ml, 23.40 +/- 6.21 and [112.73 +/- 50.03] ml) (P<0.05), while Qmax markedly lower in the former ([6.94 +/- 3.23] ml/s) than in the latter ([7.75 +/- 3.52] ml/s) (P<0.05), but PSAD showed no statistically significant difference between the two groups (0.129 +/- 0.048 vs 0.123 +/- 0.034, P>0.05). CONCLUSION: Histological prostatitis can significantly increase the PSA level, prostate volume, IPSS and PVR, and reduce the Qmax of the patient, but is not correlated with PSAD. It is an important factor influencing the clinical progression of BPH.

[Silencing effect of cell-specific RNA interference plasmid pPSMAe/p-shNS-ploy(A) loaded by transgenic vector Tf-PEG-PEI targeting nucleostemin on prostate cancer cells in vitro].

OBJECTIVE: To explore the transgenic efficiency of non-viral vector Tf-PEG-PEI and the cell specific silencing effect of plasmid pPSMAe/p-shNS-ploy(A) on prostate cancer cells. METHODS: Polyethyleneimine (PEI) was modified by using polyethylene glycol and transferrin to synthesize the non-viral vector Tf-PEG-PEI. NS-specific plasmids pPSMAe/p-shNS-ploy(A) and Tf-PEG-PEI were used to transfect prostate cancer LNCap and PC-3 cells. The changes of cell morphology, proliferation ability and cell cycle were studied after down-regulating the NS gene level. RESULTS: Tf-PEG-PEI was successfully modified. After transfection, the PSMA-expressing LNCaP cells became larger and showed more pseudopodia, having a tendency to differentiate. Their cell proliferation ability was reduced, and the detection of cell cycle showed a decrease of S phase and an increase of G(1) phase after knocking down NS gene. These targets were not changed in non-PSMA-expresing PC-3 cells. CONCLUSIONS: The non-viral vector Tf-PEG-PEI has a high ability to transfer targeted gene into target cells. The cellular specificity of short-hairpin RNA transcription driven by PSMAe/p is confirmed by silencing NS gene. The use of cell specific promoter may be an effective strategy of gene therapy for prostate cancer.

[Gene profiling after knocking-down the expression of NS gene in prostate cancer PC-3 cells].

OBJECTIVE: To screen the genes and possible signal transduction pathways involved in the mechanism of nucleostemin (NS) in the proliferation of prostate cancer. METHODS: Oligonucleotide DNA microarray was used to screen the genome changes after knocking-down expression of NS in PC-3 cells and quantitative real-time PCR was used to further confirm the important differentially expressed genes. RESULTS: 219 differentially expressed genes were found and theses genes were involved in cell cycle, cell proliferation, signal transduction, cell apoptosis and cell differentiation, etc. INK4 family genes (p15, p16, p18) were up-regulated and cyclin D1, HDAC1 were down-regulated, the main action points were CDK4/6-cyclin D and pRb-E2F1 complexes. CONCLUSION: NS may promote the progression of prostate cancer by inhibiting the expression of p15, p16, and p18 in PC-3 cells. NS is an important G(1)/S checkpoint regulator and its regulatory activity has been certified at gene level.

[Silencing nucleostemin expression reduces the proliferation of PC-3 cells].

OBJECTIVE: To detect the expression of the nucleostemin (NS) gene in prostate cancer PC-3, LNCaP and DU145 cells, and to study the effect of the NS gene on the proliferation of PC-3 cells after its silencing. METHODS: The protein and mRNA expressions of NS in PC-3, LNCaP and DU145 cells were respectively detected by immunohistochemical staining and RT-PCR. An NS-specific short-hairpin RNA (shRNA) expression plasmid was used to transfect the PC-3 cells (NS-shRNA-PC-3), followed by observation of the changes of the NS gene and the proliferation and apoptosis of the cells. RESULTS: The NS gene was highly expressed in the three types of cells. After the transfection, the NS expression and the proliferation of the NS-siRNA-PC-3 cells were remarkably reduced, while the percentage of the GO/G1 cells and the early apoptosis of the PC-3 cells obviously increased. A marked decrease was observed in the neoplasm forming ability of the NS-siRNA-PC-3 cells in the nude mice. CONCLUSION: NS is highly expressed in prostate cancer cells. The proliferation of PC-3 cells is remarkably reduced and the early apoptosis of PC-3 cells increased after silencing the NS gene by NS-specific shRNA.

[Expression of nucleostemin in prostate cancer tissues and its clinical significance].

OBJECTIVE: To explore the expression of the nucleostemin (NS) gene in prostate cancer (PCa) tissues and its clinical significance. METHODS: We detected the NS expression in PCa, benign prostatic hyperplasia (BPH) and high grade prostatic intraepithelial neoplasia (HGPIN) tissues by RT-PCR and immunohistochemistry, and analyzed the correlation between the expression of the NS protein and the clinical variables of PCa. RESULTS: The NS mRNA level was markedly higher in the PCa than in the BPH tissues. The rates of strongly positive, positive and weakly positive expressions of the NS protein were 48.8%, 36.6% and 12.2% in PCa, 4.0%, 32.0% and 56.0% in BPH, and 5.0%, 25.0% and 60.0% in HGPIN, respectively. The expression level of the NS protein was significantly higher in PCa than in BPH and HGPIN (P < 0.05). The expression of the NS gene was negatively correlated with the degree of cell differentiation in the PCa tissues, the worse the differentiation, the higher the NS expression level. CONCLUSION: The NS gene is highly expressed in PCa tissues and may have an important role in the adverse differentiation and malignant proliferation of prostate cancer.

Expression of nucleostemin in prostate cancer and its effect on the proliferation of PC-3 cells.

BACKGROUND: Nucleostemin is essential for the proliferation and survival of stem and cancer cells, but it is unknown whether this newly identified molecule is involved in prostate cancer pathogenesis. METHODS: Total RNA and protein were extracted from prostate cancer tissues and PC-3, LNCap and DU145 cell lines. The nucleostemin mRNA and protein expression were measured by RT-PCR and Western blot. Immunohistochemistry was also used to detect the nucleostemin protein expression in prostate cancer tissues and PC-3 cells. A nucleostemin specific, short hairpin RNA, expression plasmid was used to transfect PC-3 cells. The changes of nucleostemin gene were detected and the proliferative capacity of the cells was determined. RESULTS: Nucleostemin was highly expressed in prostate cancer tissues and cell lines. Nucleostemin expression level in the silencer group PC-3 cells remarkably reduced. The proliferation rate of silencer group PC-3 cells decreased and the percentage of G1 stage cells increased. The neoplasm forming capacity in nude mice of the silencer group PC-3 cells decreased significantly. CONCLUSIONS: Nucleostemin is highly expressed in prostate cancer tissues and cell lines. The proliferative capacity of PC-3 cells is remarkably reduced after silencing nucleostemin gene expression.

[Expression of prostate cancer antigen-1 in prostate cancer and its clinical significance].

OBJECTIVE: To investigate the expression of prostate cancer antigen-1 (PCA-1) in different prostate tissues and analyze its correlation with the clinical parameters of prostate cancer (PCa). METHODS: The expression of PCA-1 mRNA was detected by RT-PCR in the samples from 45 cases of PCa with various clinico-pathologic characteristics, 30 cases of high-grade prostatic intraepithelial neoplasia (HG-PIN), 43 cases of BPH and 39 cases of other carcinoma tissues. The correlation of PCA-1 mRNA expression with the clinical parameters of PCa was statistically analyzed and the PCA-1 expression was examined in different samples by immunohistochemistry. RESULTS: The positive expression rate of PCA-1 mRNA was 88.9% and 60.0% and that of PCA-1 protein was 84.4% and 50.0% in the patients with PCa and HG-PIN, respectively. PCA-1 mRNA and PCA-1 proteins were not expressed in the BPH and other carcinoma tissues. The expression of PCA-1 mRNA was unrelated with the clinical parameters of PCa (P > 0.05). CONCLUSION: It is suggested that PCA-1 is a PCa-specific gene and its expression is unrelated to the clinical parameters of PCa. It might serve as a specific biomarker for the early diagnosis of PCa.

Prostate cancer antigen-1 as a potential novel marker for prostate cancer.

AIM: To examine the expression of prostate cancer antigen-1 (PCA-1) in prostate cancer (PCa) and to validate it as a potential marker for diagnosis of PCa. METHODS: In situ hybridization analysis of PCA-1 mRNA expression was performed on 40 benign prostate hyperplasia (BPH), 16 high-grade prostatic intraepithelial neoplasm (HG-PIN), 74 PCa and 34 other malignant carcinoma specimens. The level of PCA-1 expression was semiquantitatively scored by assessing both the percentage and intensity of PCA-1 positive staining cells in the specimens. We then compared the PCA-1 expression between BPH, HG-PIN and PCa and evaluated the correlation of PCA-1 expression level with clinical parameters of PCa. RESULTS: PCA-1 mRNA was expressed in the majority of both PCa and HG-PIN specimens but not in BPH and other malignant carcinoma. The expression level of PCA-1 increased along with a high Gleason score (P < 0.05), and was unrelated to other clinical parameters of PCa (all P > 0.05). CONCLUSION: The data suggest that PCA-1 might be a novel diagnostic marker for PCa, and that increased PCA-1 expression might denote more aggressive variants of PCa.