Infiltrating mast cells promote neuroendocrine differentiation and increase docetaxel resistance of prostate cancer cells by up-regulating p21 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of infiltrating mast cells on neuroendocrine differentiation (NED) and docetaxel sensitivity of prostate cancer (PCa) cells in vitro.</p><p><b>METHODS</b>Human PCa cell lines (LNCaP and C4-2) were co-cultured with human mast cell line (HMC-1) in Transwell chambers. Androgen receptor (AR) was silenced in C4-2 cells using sh-AR lentivirus, and p21 was knocked down and overexpressed by transfecting C4-2 cells with pLKO.1-sh-p21 and pCMV-p21, respectively. The morphological changes of LNCaP and C4-2 cells were observed. MTT assay and colony formation assay were used to assess the proliferation of LNCaP and C4-2 cells. CCK8 assay was used to detect the cell viability of C4-2 cells following docetaxel trreatment. RT-qPCR and Western blotting were performed to determine the mRNA and protein expressions of neuroendocrine markers, AR and p21 in the cells.</p><p><b>RESULTS</b>Co-culture with HMC-1 cells enhanced the neuroendocrine phenotypes, inhibited the proliferation and up-regulated the expression of p21 in LNCaP and C4-2 cells. P21 positively regulated NED through a non-AR-dependent signaling pathway, while p21 knockdown partially reversed NED promoted by the mast cells. PCa cells co-cultured with HMC-1 cells showed increased resistance to docetaxel, and silencing p21 partially reversed docetaxel resistance in PCa cells.</p><p><b>CONCLUSION</b>Infiltrating mast cells up-regulates p21 to promote NED and increase docetaxel resistance in PCa cells in vitro.</p>

Quantitative and comparative proteomics analysis in clear cell renal cell carcinoma and adjacent noncancerous tissues by 2-D DIGE / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify specific protein markers for renal cell carcinoma detection and diagnosis, as well as develop new potential therapeutic targets of the disease.</p><p><b>METHODS</b>We used two-dimensional difference in-gel electrophoresis (2-D DIGE) technique conjunction with mass spectrometry (MS) for the identification of significant differentially expressed proteins between 15cases of paired clear cell renal cell carcinoma (ccRCC) and adjacent normal renal tissues. The protein spots were considered as differentially expressed if a 1.5-fold altered expression level was observed (Student's t test, P value<0.05).</p><p><b>RESULTS</b>Of the 27 differentially expressed protein spots, 26 proteins were successfully identified. 11 proteins up-regulated in renal cell carcinoma,15 proteins down-regulated. Among them Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial (ACDSB), Aldose 1-epimerase (GALM), Peroxiredoxin-4 (PRDX4), Macrophage-capping protein (CAPG), Beta-defensin 107 (D107A), Microfibril-associated glycoprotein 4 (MFAP4) were first time screening as new differential expressed proteins by protomic study in renal cell carcinoma.</p><p><b>CONCLUSIONS</b>2-D DIGE is a useful technique for screening and analysis differential expressed proteins in renal cell carcinoma. These new differently expressed proteins may be useful for development new molecular markers for the tumor.</p>

Value of MN/CAIX in the diagnosis of renal cell carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of MN/CAIX in patients with renal cell carcinoma (RCC) and assess the value of MN/CAIX in the diagnosis of RCC.</p><p><b>METHODS</b>RT-PCR was employed to detect MN/CAIX mRNA in the carcinoma tissue and peripheral blood of 62 patients with RCC, using normal renal tissue and peripheral blood sample from 32 patients without RCC as control. Immunohistochemistry was used to detect MN/CAIX protein in the tissue specimens of clear cell RCC (n=36), non-clear cell renal neoplasm (n=17) and normal kidney (n=16).</p><p><b>RESULTS</b>The positivity rate of MN/CAIX mRNA was 82.3% (51/62) in renal carcinoma tissues and 54.8% (34/62) in the peripheral blood from patients with RCC, significantly higher than the rates in the control cases (P<0.05). In cases of clear cell renal cell carcinoma, the positivity rate of MN/CAIX mRNA was 98% (49/50) in the carcinoma tissues and 66% (33/50) in the peripheral blood, significantly higher than the rates in cases of non-clear cell type of RCC (P<0.05). Immunohistochemistry showed a significantly higher positivity rate of MN/CAIX protein in clear cell RCC tissues [97.2% (35/36)] than in non-clear cell renal neoplasm and normal renal tissues (P<0.05).</p><p><b>CONCLUSION</b>MN/CAIX is specifically overexpressed in RCC, especially in clear cell RCC, suggesting its potential in the diagnosis and prognostic and therapeutic evaluation of RCC.</p>

Synchronous squamous cell carcinoma of the renal pelvis and squamous cell carcinoma of the ureter: report of two cases and review of literature / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the clinicopathological characteristics of synchronous squamous cell carcinoma (SCC) of the renal pelvis and SCC of the ureter.</p><p><b>METHODS</b>The clinical data of two cases of synchronous SCC of the renal pelvis and SCC of the ureter were retrospectively reviewed and analyzed. In case 1, a 68-year-old man with hematuria for a month, imaging modalities revealed a right renal pelvis tumor and a right distal ureter tumor. The patient underwent nephroureterectomy and excision of the bladder cuff. Case 2, a 60-year-old man with the complaint of lower abdominal pain and left flank pain for a month, was diagnosed as left distal ureteral stone in another hospital. Ureterolithotomy was performed and a ureteral tumor was found at the lower site of the stone intraoperatively. The pathological report demonstrated SCC, and the patient was transferred to our hospital for further treatment. We found a left renal mass invading the left hemicolon during surgery, and nephroureterectomy was performed with a bladder cuff excision, left hemicolon resection, and also complete lymph node dissection. Neither of patients received adjuvant radiotherapy/chemotherapy.</p><p><b>RESULTS</b>Moderately differentiated SCC was reported in both of renal pelvis and ureter in case 1 and the tumor invaded the subepithelial connective tissue in the renal pelvis and superficial muscle in the ureter. In case 2, moderately differentiated SCC of the left renal pelvis with colon metastasis and poorly differentiated SCC of the ureter was reported with two retroperitoneal lymph node metastases. The two patients died from tumor recurrence and metastasis 5 and 6 months after the surgery, respectively.</p><p><b>CONCLUSION</b>Synchronous SCC of the renal pelvis and SCC of the ureter are rare and has high likeliness of early recurrence and metastasis, often with poor prognosis.</p>

Detection of Skp2 and p27kip1 expression in human renal cell carcinoma using tissue chip technique / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of skp2 and p27kip1 in human renal cell carcinoma (RCC) using tissue chip technique, and evaluate the relationship between the proteins and the biological behavior of RCC.</p><p><b>METHODS</b>Tissue chip technique and immunohistochemical SP method was used to detect the expression of skp2 and p27kip1 in normal and tumor tissues.</p><p><b>RESULTS</b>The positivity rate of Skp2 in RCC was significantly higher than that in normal renal tissues (P=0.025). The positivity rate of Skp2 expression in RCC was significantly correlated to poor differentiation of the tumor (P=0.002), and was not associated with the patients gender, age, tumor size, lymph node metastasis and stages of RCC (P>0.05). The positivity rate of p27kip1 in RCC was significantly lower than that in normal renal tissues (P=0.007). The positivity rate of p27kip1 expression was inversely correlated to the malignancy and stage of RCC (P<0.05), but not with the patients' age, gender, lymph node metastasis and tumor size (P>0.05). An inverse correlation was noted between Skp2 and p27kip1 expressions (r= -0.273, P=0.014).</p><p><b>CONCLUSION</b>Overexpression of Skp2 protein may lead to decreased p27kip1 level in RCC, indicating its involvement in the carcinogenesis and development of RCC.</p>

Expression of human telomerase reverse transcriptase in renal cell carcinoma and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of human telomerase reverse transcriptase (hTERT) in renal cell carcinoma (RCC) and its clinical significance.</p><p><b>METHODS</b>The expression levels of hTERT mRNA and protein were detected using RT-RCR and Western blotting in 45 RCC tissues, 45 adjacent tissues and 786-0 cell line, and the associations of hTERT expression with the tumor size, clinical stage, pathological type and grade were evaluated.</p><p><b>RESULTS</b>hTERT mRNA and protein was expressed at significantly higher levels in RCC tissues than in the adjacent tissues (P=0.000), and no correlation of hTERT expression was found with the tumor size, clinical stage, pathological type or grade.</p><p><b>CONCLUSION</b>hTERT might serve as a diagnostic and prognostic marker for RCC, and also shed light on the new clues for gene therapy of RCC.</p>

Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system / 南方医科大学学报

<p><b>OBJECTIVE</b>To achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.</p><p><b>METHODS</b>The gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.</p><p><b>RESULTS</b>The plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.</p><p><b>CONCLUSION</b>Efficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.</p>