UXT, a novel DNMT3b-binding protein, promotes breast cancer progression via negatively modulating lncRNA MEG3/p53 axis.

Overexpressed ubiquitously expressed transcript (UXT) in breast tumors and derived cell lines modulated the transcriptional activity of estrogen receptor alpha. However, how UXT exerts its biological functions in the tumorigenicity of breast cancer remains largely unknown. Expressions of UXT and maternally expressed gene 3 (MEG3) were examined by qRT-PCR and Western blot. The capacity of cell proliferation, apoptosis, migration, and invasion was assessed using CCK-8, flow cytometry, and transwell assays. Methylation-specific PCR (MS-PCR) was employed to evaluate the methylation of the MEG3 imprinting control region. Co-immunoprecipitation was performed to verify the UXT/DNMT3b interaction. RNA immunoprecipitation (RIP) was subjected to assess the regulation of MEG3 on p53 activity. A xenograft tumor model was further conducted to certify the molecular mechanism. UXT was upregulated, while MEG3 was downregulated in breast cancer tissues and cell lines. UXT knockdown or MEG3 overexpression inhibited cell proliferation, promoted apoptosis, and weakened cell migration and invasion. Hypermethylation of the MEG3 imprinting control region was modulated by highly expressed DNMT3b. UXT inhibited MEG3 expression via recruiting DNMT3b to its imprinting control region. MEG3 positively regulated p53 activity. UXT negatively regulated the MEG3/p53 axis in a DNMT3b-dependent manner to promote tumor growth. UXT, a novel DNMT3b-binding protein, aggravates the progression of breast cancer through MEG3/p53 axis.

Primary abdominal cocoon with cryptorchidism: a case report.

BACKGROUND: We report a rare case of primary abdominal cocoon with bilateral cryptorchidism. CASE PRESENTATION: The patient had a history of laparoscopic surgery for bilateral cryptorchidism 6 years earlier. He was admitted to the hospital again due to intestinal obstruction. Surgery was performed on the patient after the failure of conservative treatment. The patient was diagnosed with primary abdominal cocoon. Instead of the greater omentum, many cocoon-like tissues surrounding the bowel were seen during operation. Abdominal surgery can increase the risk of intestinal adhesion, which is one of the main causes of intestinal obstruction, especially in patients with abdominal cocoon. We hypothesize that the surgery 6 years earlier to address transabdominal bilateral cryptorchidism accelerated the patient's intestinal obstruction. CONCLUSION: This case implies that it is important for urologists to evaluate whether their patients exhibit abdominal cocoon before cryptorchidism surgery, to choose better surgical methods and reduce the risks of poor prognosis.

Case report: small cell transformation and metastasis to the breast in a patient with lung adenocarcinoma following maintenance treatment with epidermal growth factor receptor tyrosine kinase inhibitors.

BACKGROUND: Breast metastasis from lung cancer has been reported, but not from SCLC that is transformed from lung adenocarcinoma during maintenance treatment with epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). Transformation to small cell lung cancer(SCLC), although uncommonly seen, has been associated with resistance to EGFR-TKI therapy in lung adenocarcinomas. CASE PRESENTATION: We describe a case of a 49-year-old man with lung adenocarcinoma harboring L858R point mutation at the exon 21 of the epidermal growth factor receptor (EGFR). During the maintenance treatment with EGFR-TKI, the patient presented with a right breast mass, which was accompanied by elevated serum neuron specific enolase (NSE) level. The histological examination of biopsies from the breast mass and enlarging lung mass revealed SCLC that was less sensitive to standard SCLC treatment. The breast tumor was positive for thyroid transcription factor-1 (TTF-1), consistent with a lung primary cancer. CONCLUSION: This is the first case report of small cell transformation and metastatic to the breast in a patient with lung adenocarcinoma following EGFR-TKI treatment. Repeat biopsy is important for evaluation of evolving genetic and histologic changes and selection of appropriate treatment. and serum NSE measurement may be useful for detection of small cell transformation in cases with resistance to EGFR-TKI therapy.

Proteomics analysis of gastric epithelial AGS cells infected with Epstein-Barr virus.

Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization- time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/ differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.

[Impact of breast density on computer-aided detection (CAD) of breast cancer].

OBJECTIVE: To evaluate the impact of breast density on computer-aided detection (CAD) for breast cancer and the CAD false-positive rate of normal controls. METHODS: Two hundred and seventy-one histologically proven breast malignant lesions (from Feb. 2008 to Dec. 2009) and 238 randomly selected normal cases were classified by mammographic density according to the American College of Radiology breast imaging reporting and data system (BI-RADS). Mammograms of BI-RADS 1 or BI-RADS 2 density were categorized as non-dense breasts, and those of BI-RADS 3 or BI-RADS 4 density were categorized as dense breasts. Full-field digital mammography (GEMS Senographe) were performed in all patients and controls with craniocaudal (CC) and mediolateral oblique (MLO) views. Then the image data were transferred to review workstation (SenoAdvantage), and the lesions were marked by Second Look Digital CAD system (version 7.2, iCAD). The differences of sensitivity and false-positive rate between dense and non-dense breasts were compared. RESULTS: Overall, the sensitivity of CAD in detection of cancers was 84.1% (228/271), there was a statistically significant difference in CAD of cancers in dense versus non-dense breasts (P = 0.015). The sensitivity of CAD in detection of mass cancers was 76.5% (186/243), in detection of calcification cancers was 79.1% (125/158), there was no statistically significant difference in CAD performance for the detection of mass cancers versus calcification cancers (P = 0.547). There was a significant difference in the CAD performance for the detection of mass cancer cases in non-dense versus dense breasts (P = 0.001), but no significant difference in the CAD for the detection of calcification cancers in non-dense versus dense breasts (P = 0.216). In the controls, the distribution of mass false-positive marks did not differ significantly between non-dense and dense breast tissue cases (P = 0.207), but the distribution of calcification false-positive marks differed significantly between non-dense and dense breast tissue cases (P = 0.001). There was a statistically significant difference of false-positive marks in non-dense versus dense breasts (P = 0.043). CONCLUSIONS: The sensitivity of CAD in the detection of breast cancers is impacted by breast density. There is a statistically significant difference in the CAD performance for the detection of cancer cases in non-dense versus dense breasts. The false-positive rate of CAD is lower in dense versus non-dense breasts. It appears difficult for CAD in the early detection of breast cancer in the absence of microcalcifications, particularly in dense breasts.

Decreased topoisomerase IIα expression and altered histone and regulatory factors of topoisomerase IIα promoter in patients with chronic benzene poisoning.

Topoisomerase IIα (Topo IIα) has been implicated in the benzene-induced hemotoxicity in vitro. This study was to examine the effect of in vivo chronic benzene exposure on Topo IIα in human bone marrow mononuclear cells, and to further explore the mechanism underlying decreased Topo IIα expression in patients with chronic benzene exposure. Topo IIα activity, expression, and mRNA level assessed by DNA cleavage/relaxation assay, Western blot, and reverse transcriptase-PCR, decreased in patients with benzene exposure. These changes were accompanied by reduced histone H4 and H3 acetylation and H3K4 methylation, and increased H3K9 methylation in the Topo IIα promoter, which were evaluated by chromatin immunoprecipitation (ChIP) assay. In addition, there were alterations in mRNA levels of Topo IIα promoter regulatory factors such as SP1, ATF-2, SP3, NF-YA, NF-M, P53, C-MYB, C-JUN, and ICBP90. Our results demonstrate that Topo IIα expression was reduced in patients with chronic benzene exposure, which was accompanied by alterations in histone acetylation and methylation and regulatory factor mRNA levels of Topo IIα promoter.

[Acquired somatic mutations in non-coding region of bcl-6 gene in germinal center-type diffuse large B-cell lymphoma].

OBJECTIVE: To investigate the mutation of 5' non-coding region of bcl-6 gene in germinal center B-cell (GCB) subtype of diffuse large B-cell lymphoma (DLBCL). METHODS: t(14;18) detection and immunohistochemical staining (EnVision method) were performed in 60 cases of DLBCL, which were divided into GCB and non-GCB subtypes. Polymerase chain reaction (PCR), single-strand conformation polymorphism and direct DNA sequencing were used to identify mutations in the 5' non-coding region of the bcl-6 gene. RESULTS: Seven of 60 cases showed t(14;18) translocation in the major breakpoint region. Using minimally acceptable criteria, 18 of 60 cases were probably to be germinal centre derived. Bcl-6 mutations were detected in 12 of 60 cases (20.0%) of DLBCL, with a significantly higher frequency in the GCB subgroups (7/18) than in the non-GCB subgroups (11.9%, 5/42). Bcl-6 mutations occurred most frequently in +363 and +469 sites. An association of bcl-6 mutation and GCB subgroup was obtained. CONCLUSIONS: The 5' regulatory region of the bcl-6 gene underwent less frequent somatic hypermutation during lymphomagenesis than the results of previous reports. Bcl-6 mutation occurred mostly in the GCB subtype and detection of t(14;18) seems helpful in the classification of DLBCL.

[Construction and identification of shRNA expressing vectors targeting RhoA gene].

AIM: To construct and identify shRNA interference vector targeting RhoA gene. METHODS: Two pairs of single stranded oligonucleotides encoding RhoA siRNA sequence were designed and synthesized. After annealing, its were inserted into vector pGPU6/GFP/Neo, constructed recombinant vectors and then were identified by restrictive digestion and DNA sequencing. LoVo cells were transfected with the recombinant DNA samples by the liposome complex method, and then fluorescence photographs were taken. RESULTS: Enzyme digestion and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pGPU6/GFP/Neo vector. CONCLUSION: The RhoA gene-targeted siRNA and its vector are successfully constructed.

Clinical significance of miR-21 expression in breast cancer: SYBR-Green I-based real-time RT-PCR study of invasive ductal carcinoma.

Growing evidence suggests microRNAs (miRNAs) have an important role in tumorigenesis. MicroRNA-21 (miR-21) is up-regulated in many malignant tumors, including breast cancer. Its association with clinicopathologic features and expression of PTEN (phosphatase and tensin homolog deleted on chromosome 10), one of its target genes, in breast cancer has not been reported systematically. To further determine the potential involvement of miR-21 in breast cancer, we have evaluated the expression level of miR-21 by stem-loop real-time RT-PCR based on SYBR-Green I in human invasive ductal carcinoma of the breast, and we have correlated the results with clinicopathologic features and PTEN protein expression. Matched non-tumor and tumor tissues of 40 human invasive ductal carcinoma of the breast were analyzed for miR-21 expression by stem-loop real-time RT-PCR based on SYBR-Green I. Immunohistochemistry (IHC) was used to estimate PTEN expression in tumor tissue. The expression levels of miR-21 were correlated with PTEN and commonly used clinicopathologic features of breast cancer. The stem-loop real-time RT-PCR based on SYBR-Green I was sensitive and specific enough to detect miR-21. Expression levels of miR-21 were significantly higher in tumor tissues than the levels in matched non-tumor tissues (P=0.000). Expression of miR-21 was negatively correlated with expression of PTEN (P=0.013). Up-regulated miR-21 expression was associated with lymph node positivity (P=0.01), higher proliferation index (ki67>10%) (P=0.03) and advanced breast cancer TNM clinical stage (P=0.021). These findings suggest that PTEN is possibly one of the targets of miR-21 in breast cancer and high expression of mir-21 indicates a more aggressive phenotype.

[Expression of microRNA-21 in invasive ductal carcinoma of the breast and its association with phosphatase and tensin homolog deleted from chromosome expression and clinicopathologic features].

OBJECTIVE: To investigate the expression of microRNA-21 (mir-21) in invasive ductal carcinoma (IDC) of the breast and its association with phosphatase and tensin homologue deleted from chromosome (PTEN)-10 protein expression and the clinicopathologic features of IDC. METHODS: Specimens of IDC and normal tissues more than 5 cm away from the tumor tissues were collected from 40 IDC patients, all female, aged 53 (35-77). Stem-loop real-time RT-PCR was used to examine the mir-21 expression. Immunohistochemistry was used to examine the PTEN protein expression in the tumor tissue. The association of mir-21 expression with the PTEN expression and the clinicopathologic features of the breast IDC were analyzed. RESULTS: Compared with the nontumor control samples, the median (M) of relative expression of mir-21 (2(-DeltadeltaCt)) was 5.770 (25th-75th percentile, 3.605-7.255) in the tumor samples, significantly higher than that of the nontumor control samples (set at 1.000, P<0.001). Reduced PTEN protein expression was seen in 45% (18/22) of all cases. The expression of mir-21 was higher in the group of reduced PTEN expression (with an M of 6.800) than in the group of high PTEN expression (with an M of 4.850, P=0.013). The up-regulated expression of mir-21 was positively correlated with the TNM clinical stage, lymph node positivity, and proliferation index (P=0.021, 0.010, and 0.030 respectively). CONCLUSION: mir-21 plays an important role in the development and progression of breast cancer. PTEN is possibly one of the targets of mir-21.

[Breast intraductal lesion resection under breast fiberoptic ductoscopy].

OBJECTIVE: To explore the feasibility and to sum up the experience of breast intraductal neoplasm resection under breast fiberoptic ductoscopy (FDS). METHODS: FDS was performed on 548 patients with nipple discharge from Sep.2004 to Nov.2006. The clinical data of breast intraductal neoplasm found by FDS in patients who underwent tumor resection were analyzed, and the breast intraductal neoplasm image characteristics, diagnosis, operative type and postoperative pathological results were analyzed. RESULTS: Of the 548 patients with nipple discharge, intraductal neoplasm was found in 187 cases (34.1%), intraductal papilloma in 159 cases (29.0%), intraductal papillomatosis in 12 cases (2.2%), and breast carcinoma in 16 cases (2.9%). One hundred thirty-five patients were operated on in our hospital, of whom 91 were performed tumor resection or segmentectomy under the localization by FDS, and the other 44 were performed segmentectomy after breast duct infusion of methylene blue. The diagnostic rate under FDS in the FDS group (97.8%) was higher than that in the breast duct infusion methylene group (86.4%) (chi2=6.96, P=0.008). CONCLUSION: FDS is not only an accurate diagnosis for breast intraductal lesion, but also an assistance to localize the breast intraductal neoplasm and to remove them in the operation.

Amifostine protects bone marrow from benzene-induced hematotoxicity in mice.

Benzene is one of the most widely used industrial chemical agents. Long-term benzene exposure causes bone marrow aplasia and leads to a wide range of hematopoietic disorders including aplastic anaemia (AA). There are currently no effective approaches to protect people from benzene-induced hematotoxicity and AA. In addition, current treatments for AA have limitations with short- and long-term risks. Protective agents and new therapeutic approaches, therefore, are needed to prevent and treat the disease. Amifostine is a well-known cytoprotective agent and has been widely used in clinical for protecting normal tissues from the toxic effects of chemotherapy and radiotherapy. The authors utilized an established mouse model to determine the protective effect of amifostine on benzene-induced bone marrow hematotoxicity. Whole-blood cell count, morphological and histopathological alterations in the bone marrow and spleen, as well as the production of inducible toxic oxidative species were examined and compared among the mouse groups. Amifostine treatment in benzene-exposed mice significantly improved blood cell counts, and morphological and histopathological signs of hematotoxicity in the bone marrow as well as in the spleen. Moreover, amifostine prevented benzene-induced bone marrow and spleen cell apoptosis and rescinded the inhibition of cell proliferation induced by benzene exposure. Finally, amifostine significantly inhibited the levels of reactive oxidative species and lipid peroxidation induced by benzene exposure. These data suggest that amifostine appears to have substantial protective effect on benzene-induced bone marrow hematotoxicity.