Cardelli M.,Laboratory of Immuno Oncology
Methods in molecular biology (Clifton, N.J.) | Year: 2011
Alu PCR is a rapid and easy-to-perform "DNA fingerprinting" technique based on the simultaneous analysis of many genomic loci flanked by Alu repetitive elements, which allows the detection of genetic polymorphisms and mutations in human and primate genomes. In the protocol described in the present chapter, two fluorochrome-labelled primers complementary to Alu sequences are used to perform the PCR, and the amplification products are then analysed by capillary electrophoresis. The resulting -complex electrophoretic pattern may show sample-to-sample variability due to insertion, deletion, or sequence change of Alu retrotransposons, or caused by length variation of the sequence interposed between two Alus. Source
Lin G.,Fujian Medical University |
Tang Z.,Laboratory of Immuno Oncology |
Tang Z.,Fujian Provincial Key Laboratory of Translational Cancer Medicine |
Ye Y.-B.,Fujian Medical University |
And 4 more authors.
Oncology Reports | Year: 2012
The relationship between KRAS and NF-κB in colorectal cancer is not clear. Western blotting was used to determine whether KRAS knockdown in SW620 cells altered the levels of NF-κB-p65 and other molecules. Furthermore, we investigated the association between the KRAS status and NF-κB expression in 167 colorectal cancers tumor tissues and their correlation with overall survival (OS) of patients with KRAS mutations and activated NF-κB. RAS, p-ERK, p-IκBα and p65 expression was decreased in SW620 cells with KRAS knockdown. The MEK inhibitor U0126 downregulated p-ERK, p-IκBα and p65 levels in SW620 cells. p65 activation in tumors with KRAS mutations was higher (50.8%) than in tumors with the wild-type KRAS gene (30.6%) (P=0.012). Compared to patients with other types of tumors, OS was lower (median 28.4 months) in patients with KRAS mutations and NF-κB activation, vs. a median of 46.3 months in patients with other types of tumors (P=0.005). NF-κB activation was reduced in SW620 cells with KRAS knockdown, possibly via the RAS-ERK-IκBα pathway. The presence of both KRAS mutations and the active form of NF-κB in CRC tumors indicates poor patient prognosis. Source
Zhao S.,Fujian Medical University |
Zhao S.,Laboratory of Immuno Oncology |
Li C.,Fujian Medical University |
Ye Y.-B.,Laboratory of Immuno Oncology |
And 2 more authors.
Laboratory Medicine | Year: 2011
Objective: Chemerin was shown to play a role in the colocalization of natural killer (NK) cells, which have an antitumor role. We aimed to determine the expression of chemerin and the relationship of chemerin expression with prognosis in patients with non-small cell lung cancer (NSCLC). Methods: We examined chemerin expression and the infiltration number of NK cells in NSCLC patients using immunohistochemistry. The association of chemerin expression with clinicopathologic characteristics and prognosis was analyzed. Results: Of the NSCLC patients, 51.85% exhibited lower expression levels of chemerin protein. The chemerin expression was significantly correlated with histological grade and the infiltration of NK cells. Non-small cell lung cancer patients with a lower chemerin expression had poorer survival rates than those with a higher expression. Multi-variable Cox regression analysis revealed that the chemerin expression level was an independent factor for prognosis. Conclusions: A greater expression of chemerin is an independent predictor of a better prognosis for patients with NSCLC. Source
Tang W.,Fujian Medical University |
Su G.,Fujian Medical University |
Su G.,Laboratory of Immuno Oncology |
Li J.,Fujian Medical University |
And 10 more authors.
International Journal of Oncology | Year: 2014
Upregulation of nuclear factor-κB (NF-κB) in colorectal carcinoma (CRC) accelerates tumor growth, whereas, irinotecan (CPT-11)-induced NF-κB activation reduces chemosensitivity and weakens the anti-colorectal cancer function itself, while proteasome inhibitors can inhibit NF-κB and improve the effect of chemotherapy. Carfilzomib (CFZ) is a novel proteasome inhibitor that has been recently approved by the FDA and is in clinical use for the treatment of multiple myeloma, but little is known about its activity against CRC. The aim of the present study was to explore whether CFZ alone or in combination with CPT-11 is effective in CRC treatment. We evaluated the novel therapeutic ability and mechanism of action of CFZ in CRC in vitro and in vivo. SW620 cells were incubated with CFZ alone or in combination with CPT-11. Cell proliferation was assessed by WST-1 and clonogenic assays, the cytotoxic interaction was assessed with a combination index (CI). Cell cycle progression was analysed with flow cytometry. Cell apoptosis was evaluated by detecting the Annexin V/propidium iodide (PI) ratio, caspase 3 and CD95 expression, and with TUNEL staining. Cell migration and invasion was determined with a wound-healing assay and a Transwell matrix penetration assay. A CRC xenograft model was established to monitor tumor growth. EMSA was used to analyse NF-κB activation and western blot analysis was used to detect the protein levels of related signaling factors. CFZ significantly inhibited the growth of SW620 cells, and had synergistic inhibitory effects with CPT-11 on survival and colony formation; possibly by inhibition of NF-κB activation, MEK/ERK and PI3K/AKT pathway factor dephosphorylation and survivin downregulation. Co-administration of CFZ and CPT-11 induced G2/M arrest, increased p21 WAF1/CIP, and decreased mutant p53 and cdc25c expression. Induction of apoptosis was accompanied by marked increases in PARP cleavage, caspase 3 activation, an increase of CD95 and p-p38, and ATF3 activation. Combination treatment lowered the invasive and migration ability of SW620 cells, reduced MMP and increased TIMP protein expression. Finally, co-administration of CFZ and CPT-11 suppressed tumor growth and increased apoptosis compared with single-agent treatment in SW620 xenograft models correlated with NF-κB downregulation. Carfilzomib alone or in combination with CPT-11 is effective against colorectal cancer through inhibition of multiple mechanisms related to NF-κB, and could be a potential novel therapy for CRC. Source
Chen Q.,Fujian Medical University |
Lin R.-B.,Fujian Medical University |
Ye Y.-B.,Laboratory of Immuno Oncology |
Fan N.-F.,Fujian Medical University |
And 6 more authors.
Medical Oncology | Year: 2010
Renal-cell carcinoma (RCC) is susceptible to immune therapy including the use of the nonmyeloablative allogeneic transplantation (NST). However, NST can produce severe toxicity, might not be appropriate for many patients with metastatic RCC. Other novel allogeneic immunotherapies are designed to induce an autologous immune response directed against the malignancy. In single- arm phase II trials, thalidomide has demonstrated a modest activity in the treatment of advanced RCC. Here we present a case report in which a patient with advanced RCC in the absence of transplant conditioning, that was receiving thalidomide, was infused with partially HLA-matched irradiated allogeneic lymphocytes. In this patient a complete response with weak acute graft-versus-host disease (GVHD) was observed. No evidence of the disease was present over the subsequent 36 months survival of the patient, suggesting that the infusions may have played a major role in the antineoplastic effect. A potential mechanism of this protocol may involve a host-versus-graft reactions-mediated antitumor effect against the malignancy. In addition, the present results suggest that a combination protocol with alternate treatment (e.g., chemotherapy) schedules merit further investigation in the management of various malignancies. © 2009 Humana Press Inc. Source