Xiaoming Jin

Expressions of putative cancer stem cell markers ABCB1, ABCG2, and CD133 are correlated with the degree of differentiation of gastric cancer

BackgroundThe present study was carried out to determine whether a quantitative relationship exists between the expressions of 3 cancer stem cell (CSC) markers and the degree of differentiation of gastric cancer.MethodsThe expressions of 3 putative CSC markers, ABCB1, ABCG2, and CD133, were detected in 90 human gastric adenocarcinoma cases by immunofluorescence assay. The differentiation statuses of 3 gastric cancer cell lines (the undifferentiated gastric cancer cell line HGC-27, the poorly differentiated gastric cancer cell line BGC-823, and the moderately-poorly differentiated gastric adenocarcinoma cell line SGC-7901) were observed and compared by performing the 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Gastric xenotransplant cancers in nude mice were constructed to compare the malignancy of the 3 variously differentiated gastric cancer cell lines. The expressions of the 3 putative CSC markers were also detected in the 3 gastric cancer cell lines in vitro by flow cytometric analysis and in the 3 gastric xenotransplant cancers in vivo by immunofluorescence staining.ResultsThe expressions of ABCB1, ABCG2, and CD133 were generally correlated with the degree of differentiation of the gastric cancers. In the human gastric adenocarcinomas and in the cancer cell lines, the expressions of ABCB1, ABCG2, and CD133 increased with the increases in the malignancy grades of the gastric cancers. In the human gastric adenocarcinomas, poorly differentiated adenocarcinoma expressed more ABCB1, ABCG2, and CD133 than well-differentiated adenocarcinoma. In addition, the expressions of ABCB1 and CD133 were higher in the diffuse type than in the intestinal type of human gastric cancers. The undifferentiated cell line HGC-27 expressed more putative CSC markers than the moderately-poorly differentiated cell line SGC-7901. Similar results were observed in the xenotransplant tumors that arose from the 3 gastric cancer cell lines.ConclusionsThe expressions of the CSC markers ABCB1, ABCG2, and CD133 differed in the gastric cancers with various degrees of differentiation, with poorly differentiated gastric cancer expressing relatively more CSC markers.

The efficacy and safety of leflunomide therapy in lupus nephritis by repeat kidney biopsy

To evaluate the clinical and pathological efficacy, and safety of leflunomide as a new immunosuppressive medicine in lupus nephritis (LN). A total of 31 patients were all determined as LN by kidney biopsy. SLE disease activity index (SLEDAI), clinical and immunological tests of these patients were performed. Meanwhile, the pathological presentation and LN activity of before and after leflunomide therapy were evaluated by repeat biopsy. The patients of LN usually have a bit response by the first or second month visit and have a good response by the third month visit after leflunomide therapy. One year later SLEDAI scores of all patients were significantly improved and 13 patients of them were transformed from complex pathological types to simple types (the transformed ratio was 41.9%). For the other patients not transformed, the pathological presentation took a favorable turn, the pathological active index (AI) of LN were significantly improved. There was not anyone relapsed or aggravated. The side effects of leflunomide were less and mild, and could be improved by symptomatic management with or without decreasing dosage. The clinical and pathological activity of LN can be apparently inhibited and the relapse can be prevented through leflunomide therapy. The side effects of leflunomide are mild and transient. Leflunomide is now a new ideal immunosuppressive medicine in the therapy of LN.

Hepatitis B virus induces G1 phase arrest by regulating cell cycle genes in HepG2.2.15 cells

BackgroundTo investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.MethodsMTT, colony formation assay and tumourigenicity in nude mice were performed to investigate the effect of HBV on the proliferative capability of host cells. In order to explore the potential mechanism, cell cycle and apoptosis were analysed. The cell cycle genes controlling the G1/S phase transition were detected by immunohistochemistry, westernblot and RT-PCR.ResultsHepG2.2.15 cells showed decreased proliferation ability compared to HepG2 cells. G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells. The phosphorylated Rb in HepG2.2.15 cells was decreased.ConclusionsOur results suggested that HBV inhibited the capability of proliferation of HepG2.2.15 cells by regulating cell cycle genes expression and inducing G1 arrest.

Hepatoma cell line HepG2.2.15 demonstrates distinct biological features compared with parental HepG2

AIM To investigate the biological features of hepatitis B virus (HBV)-transfected HepG2.2.15 cells. METHODS The cell ultrastructure, cell cycle and apoptosis, and the abilities of proliferation and invasion of HBV-transfected HepG2.2.15 and the parent HepG2 cells were examined by electron microscopy, flow cytometry, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trans-well assay. Oncogenicity of the two cell lines was compared via subcutaneous injection and orthotopic injection or implantation in nude mice, and the pathological analysis of tumor formation was performed. Two cytoskeletal proteins were detected by Western blotting. RESULTS Compared with HepG2 cells, HepG2.2.15 cells showed organelle degeneration and filopodia disappearance under electron microscope. HepG2.2.15 cells proliferated and migrated slowly in vitro, and hardly formed tumor and lung metastasis in nude mice. Flow cytometry showed that the majority of HepG2.2.15 cells were arrested in G1 phase, and apoptosis was minor in both cell lines. Furthermore, the levels of cytoskeletal proteins F-actin and Ezrin were decreased in HepG2.2.15 cells. CONCLUSION HepG2.2.15 cells demonstrated a lower proliferation and invasion ability than the HepG2 cells due to HBV transfection.

The response to interferon is influenced by hepatitis B virus genotype in vitro and in vivo.

PURPOSE To investigate the effectiveness of an interferon administration on different genotypes of hepatitis B virus (HBV) in vitro and in vivo. METHODS In vitro, we transfected plasmids carrying different HBV genotypes including recently identified new genotype I into HepG2 and HuH7 cells, then treated with standard interferon alpha (IFN-α); in vivo, we treated mice with pegylated interferon alpha (Peg-IFN-α) after injection with HBV DNA of different genotypes. The culture supernatants from cell culture and sera from mice were collected and used in hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) assays by ELISA and HBV DNA measurement by PCR. RESULTS Both in cell culture and in mouse model, it was observed that HBV genotypes A and B exhibited significantly better response to IFN-α2a or Peg-IFN-α2a in terms of reduced expression of HBsAg, HBeAg and the HBV DNA level as compared to HBV genotypes C and D. Moreover, the inhibitory effect of IFN-α2a or Peg-IFN-α2a on HBV genotype I was greater than on genotype C or D, but less than genotype A. However, there was no significant response difference between genotypes A and B, C and D, B and I, respectively. CONCLUSION The effectiveness of IFN/Peg-IFN to suppress HBV replication is dependent on different HBV genotypes. IFN/Peg-IFN is more effective on HBV genotype A or B than on genotype C, D or I. Treatment regimens are suggested to be adapted to HBV genotype.

miR-136 suppresses tumor invasion and metastasis by targeting RASAL2 in triple-negative breast cancer

MicroRNAs play an important role in the regulation of cancer migration, invasion and metastasis. Patients with triple-negative breast cancer (TNBC) have a high incidence of early relapse and metastasis; however, the molecular basis for metastasis and recurrence in these individuals remains largely unknown. Herein, we demonstrate that miR-136 is an anti-invasive microRNA in TNBC and suppresses mesenchymal invasion and metastasis. Our results demonstrated that miR-136 was downregulated in TNBC and negative correlated with the WHO grades. However, RASAL2 was identified as a functional target of miR-136, and was overexpressed in TNBC and correlates with pathological grades. Moreover, overexpression of RASAL2 in a breast cancer cell line rescued miR-136-mediated cell migration and invasion. In conclusion, these results indicate that the miR-136/RASAL2/MET axis act as a suppressor of TNBC metastasis.

Detection of viral antigens in renal tissue of glomerulonephritis patients without serological evidence of hepatitis B virus and hepatitis C virus infection.

OBJECTIVES Glomerulonephritis is an important extrahepatic manifestation of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. HBV and HCV infection may be occult, and they are often overlooked by both patients and doctors. The aim of this study was to assess the importance of HBV and HCV infection in glomerulonephritis patients with undetectable HBV surface antigen (HBsAg) and HCV antibody in serum. METHODS The HBsAg, the HBV core antigen (HBcAg), and the HCV antigen were detected using immunohistochemistry in frozen renal tissues of 500 glomerulonephritis patients without serological evidence of HBV and HCV infection. Electron microscopy was used to trace the virus particles, and clinicopathological features were also reviewed. RESULTS HBsAg or HBcAg was positive in nine out of 500 cases (9/500, 1.8%). Three cases were HBsAg-positive and another six cases were HBcAg-positive. The HCV antigen was found in eight cases (8/500, 1.6%). There was one case of HBV and HCV co-infection (1/500, 0.2%). Under electron microscopy, virus particles were found in the base membrane and cytoplasm of endotheliocytes in the glomerulus. The most common clinical manifestation was nephrotic syndrome (9/18), followed by nephritic syndrome (7/18). Membranous nephropathy was the most common pathological diagnosis (5/18), followed by mesangioproliferative glomerulonephritis (4/18) and IgA nephropathy (4/18). CONCLUSIONS Occult HBV and HCV infection might be implicated in HBV- or HCV-associated glomerulonephritis. More attention should be focused on the underlying cause.

From cell membrane to the nucleus: an emerging role of E-cadherin in gene transcriptional regulation

E‐cadherin is a well‐known mediator of cell–cell adherens junctions. However, many other functions of E‐cadherin have been reported. Collectively, the available data suggest that E‐cadherin may also act as a gene transcriptional regulator. Here, evidence supporting this claim is reviewed, and possible mechanisms of action are discussed. E‐cadherin has been shown to modulate the activity of several notable cell signalling pathways, and given that most of these pathways in turn regulate gene expression, we proposed that E‐cadherin may regulate gene transcription by affecting these pathways. Additionally, E‐cadherin has been shown to accumulate in the nucleus where documentation of an E‐cadherin fragment bound to DNA suggests that E‐cadherin may directly regulate gene transcription. In summary, from the cell membrane to the nucleus, a role for E‐cadherin in gene transcription may be emerging. Studies specifically focused on this potential role would allow for a more thorough understanding of this transmembrane glycoprotein in mediating intra‐ and intercellular activities.

Ubiquitin-specific peptidase 22 overexpression may promote cancer progression and poor prognosis in human gastric carcinoma

Ubiquitin-specific peptidase 22 (USP22) was recently identified as a new tumor cell marker, and previous studies demonstrated its expression in a variety of tumors and its correlation with tumor progression. Because tumor progression plays an important role in cancer, researchers are paying more attention to the correlation between USP22 expression and metastatic potential, resistance to chemotherapy, and patient prognosis. This study showed that USP22 is highly expressed in gastric cancer tissues, and significant differences in USP22 expression (P < 0.01) were identified between different types of gastric cancer (the highest expression was found in poorly differentiated adenocarcinomas). In addition USP22 expression was found to be correlated with the promotion of cancer evolution, tumor invasion, and lymph node metastasis. The C-myc protein was also shown to have synergistic effects with USP22 in gastric cancer tissue. On the basis of the results, USP22 expression may play an important role in gastric carcinoma tissue, particularly in precancerous lesions during the gastric cancer evolution process.

The value of decreased plasma gelsolin levels in patients with systemic lupus erythematosus and rheumatoid arthritis in diagnosis and disease activity evaluation

Plasma gelsolin, the extracellular gelsolin isoform, circulates in the blood of healthy individuals at a concentration of 200 ± 50 mg/l and plays important roles in the extracellular actin-scavenging system during tissue damage. Decreased plasma gelsolin levels have been observed in many inflammatory diseases. In the present study, the variation and potential clinical application of plasma gelsolin levels in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) were analysed. Plasma samples and clinical data were collected from informed and consenting participants: 47 SLE patients, 60 RA patients and 50 age- and gender-matched healthy individuals. Semiquantitative western blotting was used for measuring plasma gelsolin levels. The plasma gelsolin levels in patients with SLE and RA were significantly decreased compared with healthy controls (145.3 ± 40.4 versus 182.7 ± 38.3 mg/l and 100.8 ± 36 versus 182.7 ± 38.3 mg/l, p < 0.001), and plasma gelsolin levels were especially lower in RA than in SLE patients (100.8 ± 36 versus 145.3 ± 40.4 mg/L, p < 0.001). An analysis of the clinical data showed a significant negative correlation between plasma gelsolin levels and SLE Disease Activity Index (SLEDAI) scores (r = 0.659, p < 0.001) but no correlation between plasma gelsolin levels and RA disease activity score 28 (DAS28) (r = 0.076, p = 0.569). Different clinical characteristics were also observed in SLE and RA patients with normal and decreased plasma gelsolin levels.This study found significantly lower plasma gelsolin levels in patients with SLE and RA compared with healthy controls and documented a significant negative correlation between plasma gelsolin levels and SLEDAI, which suggested the potential clinical application of plasma gelsolin in SLE diagnosis and disease activity evaluation. The different clinical characteristics in SLE and RA patients with normal and decreased plasma gelsolin levels indicate differences in the basis of the diseases.