Up-Regulation of miR-26a-5p Inhibits E2F7 to Regulate the Progression of Renal Carcinoma Cells.

BACKGROUND: Metastasis is the main cause of renal cell carcinoma (RCC) tumor death, and effective inhibition of RCC metastasis is an essential means to meliorate the prognosis of RCC patients. MicroRNAs (miRs) have been proved to be stable and important biomarkers for several malignancies. This study is therefore set out to explore the metastasis-related miR and its mechanism in RCC. METHODS: The expression of miR- 26a -5p in RCC was analyzed using the expression profile in the Cancer Genome Atlas (TCGA). MiR-26a-5p and E2F transcription factor 7 (E2F7) in RCC patients were detected by qRT-PCR. Cell Counting Kit-8 (CCK-8) was adopted to assess cell proliferation, Transwell was utilized to evaluate migration and invasion, and flow cytometry (FC) was used to determine apoptosis. Mouse cell-derived and patient-derived xenotransplantation models were established to evaluate the effect of miR-26a-5p on tumor growth and metastasis in vivo. The molecular mechanism of miR-26a-5p was analyzed by dual-luciferase reporter (DLR) gene analysis, qRT-PCR, and Western blot (WB) both in vivo and in vitro. RESULTS: MiR-26a-5p was reduced in renal carcinoma cells and may serve as a biomarker for renal cancer metastasis and prognosis. MiR-26a-5p up-regulation inhibited migration and invasion in renal cell lines and tumor metastasis in vivo. Bioinformatics target prediction and RNA-seq results showed that E2F7 was among the targets of miR-26a-5p and was significantly inhibited by miR-26a-5p in vivo and in vitro. CONCLUSION: MiR-26a-5p presents low expression in RCC and promotes RCC cell apoptosis and prevents cells from proliferating and invading by targeting E2F7, which is a promising therapeutic target for RCC.

Effectiveness of allogeneic CD3AK cells on transplanted human renal cell cancer in mice with severe combined immune deficiency.

PURPOSE: To assess the activity of allogeneic anti-CD3 antibody induced activated killer (CD3AK) cells on transplanted human renal cell cancer (RCC) in mice with severe combined immune deficiency (SCID), thus to provide theoretical and experimental support for clinical application of allogeneic CD3AK cells in the treatment of RCC. METHODS: A culture system which can massively increase allogeneic CD3AK cells was constructed. CCK-8 method was used to detect lethal effect of allogeneic CD3AK cells on human OS-RC-2 renal cancer cell line. Then, tumor-bearing mice models were constructed. SCID mice were randomly divided into four groups: group A (caudal vein was injected with allogeneic CD3AK cells before tumor bearing), group B (the control group of group A: caudal vein was injected with PBS before tumor bearing), group C (caudal vein was injected with allogeneic CD3AK cells after tumor bearing) and group D (the control group of group C: caudal vein was injected with PBS after tumor bearing), and spleen parameters were calculated to observe any inhibitory effect of allogeneic CD3AK cells on the growth of renal cancer cells, as well as their effect on the immune system of mice. RESULTS: Compared with the control groups B and D, spleen parameters of groups A and C increased significantly (p<0.05); compared with group C, spleen parameters of group A showed no obvious difference (p>0.05); compared with the control groups B and D, tumor weight of groups A and C decreased significantly and tumors grew slowly (p<0.05); the weight of mice in all groups increased while the weight of mice in the experimental groups increased less than the weight of mice in the control groups (p<0.05); compared with group C, tumor weight of group A reduced and grew slowly (p<0.05). Immunohistochemistry revealed that CD3+ cells showed positive expression in tumor tissues of mice in group C, while no positive expression was found in tumor tissues of mice in other groups. CONCLUSION: Allogeneic CD3AK cells exerted lethal effect on OS-RC-2 cell line and significant inhibitory activity on the growth of transplanted cancer in mice with SCID. Also CD3AK cells expressed certain preventive effect on the development of implanted cancer in SCID mice; allogeneic CD3AK cells possessed antitumor activity and could enhance the immunologic functions of SCID mice with human renal cell-bearing cancer.

Bioinformatic analysis of c-Myc target from laryngeal cancer cell gene of laryngeal cancer.

AIM OF STUDY: The aim was to explore the structure and functions of new target spot c-Myc target from laryngeal cancer cell. (MTLC) of c-Myc gene. METHODS: This study adopted bioinformatic methods to analyze the physicochemical property, secondary structure, hydrophobic region, a transmembrane region, and prediction of functions. RESULTS: The results showed that the whole length of the open reading frames was 708 bp, coding was 235 amino acids. This protein was a basic protein possessed two transmembrane structures and weight was about 26592.5 Da. The main elements of secondary structure were alpha-helix and random coil. MTLC was a membrane constitutive protein that possessed signal transduction and regulation may locate on karyotheca as results of subcellular localization and function prediction. CONCLUSION: This study has provided the theoretical basis for the further discussion of the effect and mechanism of action of MTLC in the occurrence of laryngeal cancer.

Relationship between gene polymorphisms and prostate cancer risk.

OBJECTIVE: To investigate the relationship between genetic factor and prostate cancer (Pca) risk and the possible cause in it. METHODS: The polymorphisms of cytochrome P450 family 17 (CYPl7) rs743572, p27 V109G and androgen receptor (AR) gene CAG repeat length in peripheral blood from 70 cases and 70 controls were detected through the polymerase chain reaction-restriction fragment length polymorphism technique or short tandem repeat-polymerase chain reaction technique. Then, according to the results of case-control study, the recombinant plasmids containing the wild/mutant p27 gene were constructed and transfected Pca LNcap cells. After 24 and 72 h of transfection, the cell proliferative activity was determined by MTT method, cell cycle distribution and apoptosis was detected by flow cytometry, and the expression level of bcl-2, caspase-3 and p27 protein was determined by Western-blot. RESULTS: In three target polymorphisms, only p27 V109G polymorphism was related to Pca risk (P = 0.030, OR = 0.202, 95% CI = 0.042-0.973). Pca risk of p27-109G allele was lower than -109V allele (P = 0.006, OR = 0.285, 95% CI = 0.110-0.737). Cells transfected with wild/mutant p27 gene both showed the higher cells apoptosis rate and the lower cell proliferative activity than mock cells (P < 0.05 or 0.01), the regulatory effect of mutant p27 on cell proliferation and apoptosis was stronger than the wild p27 (P < 0.05). CONCLUSIONS: p27-109G allele that could cause higher p27 protein expression than -109V allele in LNcap cells, maybe is the protective factor of Pca.

Effects of different interchain linkers on biological activity of an anti-prostate cancer single-chain bispecific antibody.

BACKGROUND: A single-chain bispecific antibody (scBsAb; an engineered antibody), has promising clinical applications. Nonetheless, the effect of different interchain linkers on its activity is poorly understood. METHODS: Gene synthesis was used to splice the anti-γ-seminoprotein single-chain antibody (anti-γ-Sm scFv) gene with the anti-CD3 single-chain antibody (anti-CD3 scFv) gene via different interchain peptide linkers. The Phyre2 software was used to predict spatial configuration of different scBsAbs. Eukaryotic expression vectors carrying scBsAbs were constructed by molecular cloning techniques and these plasmids were transfected into HeLa cells with liposomes. scBsAbs were purified by Ni(2+)-NTA agarose and analysed for antigen binding by an enzyme-linked immunosorbent assay (ELISA). Blood pharmacokinetics and inhibition of prostate tumour growth in nude mice were analysed in in vivo experiments. RESULTS: Bioinformatics analysis and prediction showed that none of the three linkers, Fc, 205C', and HSA, had a significant effect on protein folding of anti-γ-Sm scFv or anti-CD3 scFv. Nevertheless, the spatial structures of the three linkers were noticeably different. Anti-γ-Sm × anti-CD3 scBsAb with an Fc, 205C', or HSA linker was successfully constructed, and these antibodies had similar protein expression levels. ELISA showed that all the three scBsAbs bound to Jurkat cells and the LNCaP membrane antigen, although binding of (205C')scBsAb was weaker than that of the two parental scFvs (P < 0.05). In contrast, binding strength of (HSA)scBsAb and (Fc)scBsAb was close to that of the parental scFvs (P > 0.05). Pharmacokinetic analysis showed that the half-clearance time of the elimination phase (T(1/2ß)) for (HSA)scBsAb was the longest: up to 4.4 h. Compared with γ-Sm ScFv, the three scBsAbs all had a much stronger inhibitory effect on the growth of prostate cancer (P < 0.05), but there were no significant differences among the three scBsAbs (P > 0.05). CONCLUSIONS: HSA is the optimal linker for the anti-γ-Sm × anti-CD3 scBsAb and may improve antigen-binding affinity of antibodies and prolong physiological retention time. Interchain linkers affect the function of scBsAbs; these effects may have important implications for construction of antibodies.

Dual specificity phosphatase 6 suppresses the growth and metastasis of prostate cancer cells.

Dual specificity phosphatase 6 (DUSP6) is expressed at low levels in numerous types of human cancer. The loss of DUSP6 plays a pivotal role in tumor progression; however, the role of DUSP6 in prostate cancer remains unclear. In this study, in vitro invasion assays and in vivo metastasis experiments were used to investigate the effects of DUSP6 on prostate cancer cell invasion and metastasis. Furthermore, in vitro growth and soft agar assays and in vivo growth experiments were performed to determine the function of DUSP6 in cell proliferation. The results showed that the overexpression of DUSP6 suppressed the invasion and growth of DU­145 human prostate cancer cells, whereas knockdown of DUSP6 promoted the invasion and proliferation of LNCap human prostate adenocarcinoma cells. Further experiments demonstrated that the overexpression of DUSP6 inhibited the proliferation and liver metastasis of DU­145 cells in mice. In addition, DUSP6 downregulated the expression of matrix metallopeptidase 3 and interleukin 8 in prostate cancer cells. Taken together, these findings indicate that DUSP6 may act as a negative mediator in the regulation of prostate cancer cell growth and metastasis.

Overexpression of oxidored-nitro domain containing protein 1 induces growth inhibition and apoptosis in human prostate cancer PC3 cells.

Previous studies have reported that oxidored-nitro domain containing protein 1 (NOR1) is a novel tumor suppressor gene identified in various types of cancer, such as nasopharyngeal carcinoma and cervical cancer. Recent studies have shown that NOR1 expression is lower in prostate cancer compared with normal prostate tissue. However, the specific function and exact mechanism of NOR1 in prostate cancer remains to be clarified. The present study aimed to investigate the function and mechanism of NOR1 in prostate cancer PC3 cells. DU145 and PC3 cells were transduced with a vector and cell viability, proliferation and apoptosis were determined. As predicted, NOR1 overexpression significantly inhibited growth and apoptosis in PC3 cells. NOR1 overexpression decreased the expression of the anti-apoptotic genes Bcl-2 and Bcl-xl and increased the level of the pro-apoptotic genes Bax and Bak in PC3 cells. Further investigation demonstrated that NOR1 overexpression activates caspase-3. Silencing of NOR1 did not inhibit growth or induce apoptosis in PC3 cells. Moreover, NOR1 inhibited proliferation and induced apoptosis via the activation of MAPK. The overexpression of NOR1 significantly inhibited tumor growth in PC3 tumor-bearing nude mice. The results suggest that the upregulated NOR1 expression was able to inhibit the progression of prostate cancer. Thus, NOR1 may be an ideal target for the treatment of prostate cancer.

[Expression and clinical significance of transcription factors in prostate cancer].

OBJECTIVE: To explore the roles of various transcription factors in the occurrence and development of prostate cancer. METHODS: A total of 139 specimens with prostate cancer (PCa) and 83 specimens with benign prostatic hyperplasia (BPH) from hospitalized patients at our hospital from 2008 to 2011 were enrolled. The mRNA expressions of c-Myc, Klf4, Nanog, Oct4A and Sox2 were determined by quantitative real-time polymerase chain reaction (PCR) and the expressions of Klf4 isoforms by conventional PCR. Immunohistochemical method was used for the detection of Klf4 protein via tissue microarray in 404 prostate samples. RESULTS: No significant difference existed in the expressions of Nanog, Oct4A and Sox2 genes between BPH and PCa samples. And the expressions of c-Myc and Klf4 genes were significantly higher in PCa than those in BPH specimens. Immunohistochemical results showed that Klf4 protein could be detected in a large majority of epithelial prostatic cells irrespective of malignant transformation. However, it was predominantly located cytoplasmically in PCa tissues and remained consistent with the expression of a differentially spliced Klf4α isoform. CONCLUSION: Klf4 is highly expressed in both BPH and PCa tissues. But in malignant cells, a specific gene product Klf4α is predominantly detectable in cytoplasm. The positioning of Klf4 protein may have an important relationship with its role in the tumorigenic process.

[Clinical significance of serum telomerase mRNA in patients with prostate cancer].

OBJECTIVE: To explore the clinical significance of detecting serum mRNA of telomerase in the diagnosis of prostate cancer. METHODS: Serum samples were collected from 29 patients with prostate cancer and 29 age-matched patients with cardiovascular or metabolic disease as non-tumor controls. Sera samples from 15 healthy age-matched subjects were used as healthy control. Detection of serum telomerase mRNA was performed with real-time reverse transcriptase quantitative polymerase chain reaction (PCR). RESULTS: Serum telomerase mRNA was detectable in 89.7% (26/29) patients with prostate cancer, but almost undetectable in non-tumor (6.9%, 2/29) and healthy control groups(1/14). CONCLUSION: Using real-time PCR for detecting serum telomerase mRNA may be an auxiliary method for diagnosing and monitoring of prostate cancer.

Study and clinical application of the anatomy of renal pedicles in retroperitoneoscopic nephrectomy / Estudio y aplicación clínica de la anatomía de los pedículos renales en la nefrectomía retroperitoneoscópica

This work aims to investigate the anatomical basis and clinical application value of renal pedicle locating in retroperitoneoscopic nephrectomy. To summarize the anatomical basis of renal pedicle locating through retrospective analysis of 278 cases of retroperitoneoscopic nephrectomy from July 2007 to September 2009, during which renal pedicle was located at about 2-4 cm below the medial arcuate ligament of the diaphragm in the space between the psoas major muscle and inferior vena cava (abdominal aorta) in the anatomical level of space before psoas. The operation of 278 patients was all successfully completed, where renal pedicle was quickly found. It took 3.5+/-1.3 min to locate the renal pedicle, and 95.6+/-23.8 min to operate. In retroperitoneoscopic nephrectomy, it is most preferable to locate renal pedicle in the space before psoas. The renal pedicle is located exactly at about 2-4 cm below the medial arcuate ligament of the diaphragm in the space between the psoas major muscle and inferior vena cava (abdominal aorta). The time for locating the renal pedicle can be shortened if the surgeon is familiar with the anatomic features of renal pedicle in retroperitoneoscopy, thereby saving the operation time.

El objetivo del estudio fue investigar las bases anatómicas y el valor de la aplicación clínica de la localización del pedículo renal en la nefrectomía retroperitoneoscópica. Para resumir las bases anatómicas de la localizacion del pedículo renal se realizó el análisis retrospectivo de 278 casos de nefrectomía retroperitoneoscópica desde Julio del 2007 a Septiembre del 2009. El pedículo renal se encontró a unos 2-4 cm por debajo del ligamento arqueado medial del diafragma en el espacio entre el músculo psoas mayor y vena cava inferior (o parte abdominal de la aorta) en el nivel anatómico del espacio anterior al músculo psoas mayor. La cirugía de los 278 pacientes fue completada exitosamente, encontrándose rápidamente el pedículo renal. El procedimiento para localizar el pedículo renal tomó 3,5+/-1,3 minutos y la cirugía completa 95.6+/-23.8 minutos. En la nefrectomía retroperitoneoscópica, es preferible localizar el pedículo renal en el espacio anterior al músculo psoas mayor. El pedículo renal se encuentra alrededor de 2-4 cm por debajo del ligamento arqueado medial de la membrana en el espacio entre el músculo psoas mayor y vena cava inferior (parte abdominal de la aorta). El tiempo para localizar el pedículo renal se puede disminuir si el cirujano está familiarizado con las características anatómicas del pedículo renal en la retroperitoneoscopía, ahorrando así el tiempo total de la cirugía.

[Effects of Qiangjing Capsule on the oxidative and antioxidative system in the epididymis of varicocele rats].

OBJECTIVE: To observe the effects of Qiangjing Capsule (QC) on the oxidative amd antioxidative system in the epididymis of varicocele rats in comparison with those of Shaofuzhuyu Capsule (SC) and Wuziyanzong Capsule (WC), and to explore its possible mechanism of enhancing epididymal sperm maturation. METHODS: Ten of 100 adolescent male SD rats were randomized to a sham-operation group, and varicocele models were successfully established in 72 of the other 90 by narrowing of the left renal vein. Then the model rats were equally assigned to 6 groups: model control, high-dose QC (0.216 g/ml), medium-dose QC (0.108 g/ml), low-dose QC (0.054 g/ml), SC (0.146 g/ml), and WC (0.130 g/ml). After 4 weeks of treatment, we determined the activity of glutathione peroxidase (GPx) and the level of malondialdehyde (MDA) in the left epididymis of different groups of rats. RESULTS: Compared with the sham-operation group, the model group showed a significant decrease in GPx activity (P < 0.01) and a marked increase in the MDA level (P < 0.05). And the high-dose QC group exhibited a significantly hither GPx activity and lower MDA level than all the other groups (P < 0.01 and P < 0.05). CONCLUSION: Varicocele can reduce the activity of GPx and elevate the level of MDA in the epididymis of rats, while Qiangjing Capsule can increase the former and decrease the latter, and thereby may improve epididymal microenvironment, enhance epididymal sperm maturation and promote fertility.

[Anatomical path of intraperitoneal radical nephrectomy: clinical experience of 60 cases].

OBJECTIVE: To evaluate the safety and efficacy of intraperitoneal anatomical radical nephrectomy (IARN). METHODS: A retrospective analysis was performed for 60 consecutive patients undergoing IARN at our hospital from March 2007 to December 2009. Various clinical parameters were collected and analyzed statistically. RESULTS: Sixty operations were performed successfully. There was neither conversion into open surgery nor blood transfusion. The mean operative time was (106 ± 23) min, mean intraoperative estimated blood loss (112 ± 37) ml, mean time of resuming oral intake (2.1 ± 0.7) d, mean time to ambulation (1.9 ± 1.1) d, mean postoperative analgesics (pethidine) dosage (65 ± 25) mg, average drainage volume 100 (50 - 300) ml, mean time of extracting drainage tube (3.6 ± 1.3) d and mean postoperative hospital stay (9.4 ± 2.1) d. CONCLUSION: IARN offers the advantages of distinct anatomical level, shorter operative time, less hemorrhage, less damage, faster postoperative recovery and a lower rate of complications.