Expression of phosphoglycerate kinase 1 in endometrial carcinoma and its association with patients' outcome / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of phosphoglycerate kinase 1 (PGK1) and its prognostic value in endometrial carcinoma (EC).</p><p><b>METHODS</b>The expression of PGK1 was detected immunohistochemically in 30 normal endometrium and 130 EC specimens. The relationship between PGK1 protein expression and the clinicopathological features of the patients was evaluated.</p><p><b>RESULTS</b>Immunohistochemical analysis revealed low PGK1 expression in 55.4% (72/130) and high PGK1 expression in 44.6% (58/130) of the EC specimens, as compared with the rates of 90% (27/30) and 10% (3/30) in normal endometrium, respectively (P<0.001). PGK1 expression was significantly correlated with FIGO stage (P<0.001), histological grade (P=0.002) and lymph node metastasis (P<0.001). Kaplan-Meier survival analysis indicated that patients with a high PGK1 expression had a shorter overall survival rate than those with a low PGK1 expression (P<0.001). Multivariate analysis showed that a high PGK1 expression was not the independent predictor of the prognosis of EC (P=0.077).</p><p><b>CONCLUSION</b>A high expression of PGK1 is associated with aggressive and metastatic behaviors of EC, and detection of PGK1 provides assistance in evaluating the prognosis of patients with EC.</p>

Coexpression of MAP2K4 and vimentin proteins in human endometrial carcinoma and its clinicopathological significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the expression of MAP2K4 and vimentin in human endometrial carcinoma (EC) and their association with the clinicopathological features and prognosis of the patients.</p><p><b>METHODS</b>MAP2K4 and vimentin expressions were detected immunohistochemically in paraffin-embedded tissue sections from 128 patients with EC, and the correlation of MAP2K4 and vimentin expressions with the clinicopathological factors of the patients was analyzed.</p><p><b>RESULTS</b>MAP2K4 and vimentin proteins were positively expressed in 49 (38.3%) and 83 (64.8%) of the patients, respectively. A positive expression of MAP2K4 was negatively correlated with FIGO stage of the tumor (P=0.010) and lymph node status (P=0.016); a positive expression of vimentin was positively correlated with FIGO stage of the tumor (P=0.025), histological grades (P=0.017), depth of myometrial invasion (P=0.044) and lymph node status (P=0.032). MAP2K4 was inversely associated with vimentin expression in EC(r=-0.598, P<0.001). Patients positive for MAP2K4 tended to have a higher overall survival rate (P=0.002), and those positive for vimentin tended to have a lower overall survival rate (P=0.007); patients positive for MAP2K4 but negative for vimentin had the longest survival time, while those negative for MAP2K4 and positive for vimentin had lowest survival rate (P=0.004).</p><p><b>CONCLUSION</b>Detection of MAP2K4 and vimentin might help in early diagnosis and prognostic evaluation of patients with EC.</p>

Construction of a lentiviral vector containing human NESG1 gene and its expression in 293FT cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector carrying human NESG1-EGFP gene and observe its expression in 293FT cells.</p><p><b>METHODS</b>The CDS region of NESG1 gene was amplified from a plasmid containing the full-length NESG1 sequence and cloned into the lentiviral vector pGC-FU-EGFP by restriction endonuclease AgeI digestion and T(4) DNA ligase ligation. After transformation into competent E. coli cells, the candidate clones were identified by PCR and sequencing. The recombinant plasmid and the two packaging plasmids were co-transfected into human embryonic kidney cell line 293FT cells by lipofectamine 2000 to produce the lentiviral particles, and the viral titer was determined. The 293FT cells were infected by the lentiviral particles obtained and the transfection efficiency was assessed under fluorescent microscope. Western blotting was used to detect the expression of NESG1 protein in the transfected cells.</p><p><b>RESULTS</b>The lentiviral vector pGC-FU-NESG1-EGFP for NESG1 gene was constructed successfully. Strong green fluorescence was observed in 293FT cells under fluorescent microscope after co-transfection of the cells with the 3 plasmids of lentiviral vector. The virus in the supernatant reached a titer of 2×10(7) TU/ml. The transfection efficiency of the collected virus exceeded 90% in 293FT cells with a multiplicity of infection of 1. Western blotting identified the presence of NESG1 expression in the transfected 293FT cells.</p><p><b>CONCLUSION</b>The lentiviral vector for NESG1 has been successfully constructed with a high yield of lentivirus, which facilitate further investigation of the roles of NESG1 gene in the development and progression of nasopharyngeal carcinoma.</p>

Relationship between MAP3K5 and Epstein-Barr virus-encoded miR-BART22 expression in nasopharyngeal carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of MAP3K5 and miR-BART22 encoded by Epstein-Barr virus and explore their relationship in nasopharyngeal carcinomas (NPCs).</p><p><b>METHODS</b>Fifty-three archived specimens of NPCs and 30 nasopharyngitis specimens were collected for detecting the expression of EBERs and miR-BART22 by in situ hybridization, and the expression of MAP3K5 was detected using immunohistochemistry. Ten fresh NPC and 10 fresh nasopharyngitis specimens were also obtained for determining the protein expression of MAP3K5 by Western blotting.</p><p><b>RESULTS</b>EBERs were positive in all the 53 NPC specimens, and miR-BART22 was positive in 49 specimens; all the 30 nasopharyngitis specimens were negative for EBER or miR-BART22. In the 53 NPC tissues, 50 were negative for MAP3K5 expression in the cancer areas but positive in the adjacent mucosal areas, with the other 3 specimens showing a weak positivity (+). In the 30 nasopharyngitis specimens, 25 showed strong MAP3K5 positivity, 3 showed weak positivity and 2 were negative for MAP3K5 (P<0.001). Western blotting showed that the expression of MAP3K5 protein was significantly higher in nasopharyngitis than in NPC tissues (P=0.029). The expression of MAP3K5 and miR-BART22 was inversely correlated (P<0.001).</p><p><b>CONCLUSION</b>Compared with the adjacent mucosal tissues, NPC tissues have a lower expression of MAP3K5 but a higher expression of miR-BART22. The expression of MAP3K5 and miR-BART22 is inversely correlated, suggesting the possibility of MAP3K5 to serve as target gene of EBV miR-BART22. miR-BART22 may inhibit the expression of MAP3K5, thus reducing the protein phosphorylation of MAPK pathway downstream genes, inhibiting NPC cell apoptosis, preventing their differentiation and promoting their escape from immune surveillance.</p>

Isolation and detection of label-retaining cells in a nasopharyngeal carcinoma cell line / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Detection of label retaining cells (LRCs) has been a method to confirm existence of stem cells, and bromodeoxyuridine (BrdU) has commonly been used for labeling. In this study, to verify stem cells in nasopharyngeal carcinoma (NPC), LRCs were established and detected in NPC cell line 5-8F.</p><p><b>METHODS</b>The 5-8F cells were cultured with BrdU and inoculated subcutaneously into nude mice. By immunohistochemistry, immunocytochemistry, and immunofluorescence, BrdU was detected in 5-8F cells and xenograft tumors.</p><p><b>RESULTS</b>BrdU was strongly positive in cells on the 2nd and the 7th day after being added BrdU, while negative when cells were cultured without BrdU. However, only sporadic cells were positive on the 14th day after BrdU being washed out, and these cells were thought to be LRCs. The average percentage of LRCs was (0.67 +/- 0.32)%. After being cultivated with BrdU for 48 h, 5-8F cells were inoculated into nude mice subcutaneously. After chasing 8 weeks, only sporadic LRCs were detected in xenograft tumors, with a proportion of (0.55 +/- 0.36)%, and these LRCs were located at cancer margin.</p><p><b>CONCLUSION</b>The existence of LRCs in 5-8F cells indicates the existence of cancer stem cells in NPC.</p>

Dynamic contrast-enhanced CT patterns and pathologies of noncalcified pulmonary tuberculomas / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze characteristic CT enhancement patterns of noncalcified pulmonary tuberculomas and their pathological basis.</p><p><b>METHOD</b>Fifty-six patients with noncalcified pulmonary tuberculomas underwent surgical resection of the tuberculomas. Enhanced CT images of these tuberculomas were reviewed and analyzed in relation to the histological findings.</p><p><b>RESULTS</b>Of the 56 patients, 45 showed no enhancement in the tuberculomas, which were histologically characterized by central caseous necrosis and a poorly vascularized peripheral fibrotic zone. Eleven patients showed ring-like or eggshell enhancement, and the central low density region was histologically confirmed to be caused by caseous or liquefied necrosis, while the ring enhancement resulted pathologically from moderately or well vascularized peripheral fibrotic or granulomatous tissues.</p><p><b>CONCLUSIONS</b>Pulmonary tuberculomas consists mainly of caseous necrotic tissues characterized by no enhancement and ring or eggshell enhancement on dynamic contrast-enhanced CT.</p>

Construction and identification of a recombinant lentivirus vector of latent membrane protein 1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentivirus vector of latent membrane protein 1 (LMP1) and detect the expression of LMP1 in vitro.</p><p><b>METHODS</b>The LMP1 fragment including all the exons was amplified by PCR and inserted to the downstream of CMV promoter in the lentivirus vector pCDF. The three plasmids (packaging plasmid pFIV-34N, envelope plasmid pVSV-G and target plasmid pCDF-LMP1) were packaged into 293FT cells via liposome. The virus supernatant was harvested, concentrated and titrated. Mouse B lymphoma cell line A20 was transfected with the recombinant lentivirus vector of LMP1, and the expression of LMP1 in A20 cells was detected by RT-PCR and Western blotting.</p><p><b>RESULTS</b>DNA sequencing confirmed that the sequence of PCR-amplified LMP1 was consistent with the GenBank data. The LMP1 gene fragment was cloned into pCDF in the right direction, and the open reading frame of LMP1 was maintained. The 3 plasmids were effectively transferred into 293FT cells, which emitted green fluorescence in the cytoplasm and on the cell membrane under fluorescence microscope. The titer of the lentivirus vector reached 10(7) Tu/ml with a transfection efficiency 90% in A20 cells. LMP1 expression was detected by RT-PCR and Western blotting in transfected A20 cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector of LMP1 constructed can be effectively transfected into A20 cells, which provides a basis for exploring the role of LMP1 in the pathogenesis of lymphoma.</p>