jcru
Editor’s Choice : Factors Influencing Percentage Yield of Side Population Isolated in Ovarian Cancer Cell LineSK-OV-3
|
|
Abstract: Isolation of side population (SP) cells has been recognized as a useful technique for the isolation and identification of hematopoietic stem cells or cancer stem cells (CSCs). Thus the yield and purity of isolated SP cells would have a profound influence on the research outcomes in these two important areas. Hoechst 33342 exclusion assay technique has been used for the identification of SP cells. However, diverse Hoechst staining protocols giving different SP yields even from the same tissue type or same cell line have been reported in different laboratories. In this study we systematically investigated the underlying factors influencing the SP yield using Hoechst dye staining and a robust platform of flow cytometric analysis of the human ovarian cancer cell line SK-OV-3. Our study revealed that SP yield was not only affected by the Hoechst 33342 concentration, staining cell density, staining cell viability, staining duration, staining medium, flow cytometric setting and SP gating strategy, but was also affected by the cell passage number in SK-OV-3. This is the first systematic study on the factors affecting SP yield in adherent cells that mimic many solid tumour tissues. Our results provide important technical guidelines to help ensure reproducible and comparable results in SP and CSCs study. Keywords: Side population (SP), SK-OV-3, cancer stem cells (CSCs), flow cytometry (FCM), Hoechst 33342.Download Full Article |
Editor’s Choice : Is the Neoadjuvant Docetaxel, Cisplatin and 5-Fluorouracil Regimen Superior to Classic Cisplatin and 5-Fluorouracil for Locoregionally Advanced Nasopharyngeal Carcinoma?
|
|
Abstract: Objectives: We retrospectively compared the toxicity and efficacy of two neoadjuvant chemotherapy regimens (docetaxel+cisplatin+5-fluorouracil vs. cisplatin+5-fluorouracil) followed by chemoradiotherapy for locoregionally advanced nasopharyngeal carcinoma. Patients and Materials: We analysed 135 patients with stage III and IVA-B nasopharyngeal carcinoma. Forty-four patients were treated with docetaxel+cisplatin+5-fluorouracil and chemoradiotherapy (TPF group), and 91 were treated with cisplatin+5-fluorouracil and chemoradiotherapy (PF group). Chemoradiotherapy was administered with weekly cisplatin. Radical radiotherapy with total doses of 70–74Gy was administered using a conventional technique, over 7 weeks in 2.0Gy/fraction; boost doses of 6–10Gy were administered in 55.6% patients (n=75) with locally advanced cancer. Results: The median follow-up was 46.5 months (range, 9.8–62.8 months), and the follow-up rate was 95%. The TPF group had better 5-year estimated progression-free survival (77.0% vs. 73.5%; P = 0.510) and overall survival than the PF group (80.7% vs. 77.9%, P = 0.446); however, there was no statistically significant difference between the groups. Toxicities in the two groups were similar; grade 3/4 oral mucositis was more common in the TPF group (27.3%) than in the PF group (15.3%) during chemoradiotherapy. Conclusions: The neoadjuvant docetaxel+cisplatin+5-fluorouracil chemotherapy led to satisfactory long-term survival and slight improvement in progression-free survival and overall survival as compared with the classic cisplatin+5-fluorouracil regimen; toxicity was tolerable. However, prospective trials are needed to prove whether docetaxel+cisplatin+5-fluorouracil is a substitute for cisplatin+5-fluorouracil. Keywords: Nasopharyngeal carcinoma, induction-concurrent chemotherapy, docetaxel, cisplatin, 5-fluorouracil.Download Full Article |
Editor’s Choice : NACA (Nascent-Polypeptide-Associated Complex α Subunit) Against Apoptosis in B Lymphoma Cell is Independent of β Subunit (NACB)
|
|
Abstract: We found depletion of NACA in two kinds of B lymphoma cell lines, Raji and Kapas, were able to induce apoptosis in this study. We also explored whether depletion of β subunit had the same effect, and we were interested in which domain of NACA was potentially responsible to this anti-apoptosis function. Lentivirus-based shRNA was used to deplete endogenous NACA or NACB. Those cells’ viabilities were measured by Alamar-blueTM assay. Cell apoptosis was identified by molecular markers caspase9 and PARP, as well as cellular markers Annexin V and propidium iodide (PI) staining. NACA mutants were constructed by PCR site-directed mutagenesis and delivered into cells by Lentivirus. Immunofluoresce was used to investigate cellular distribution in 293FT cells. Our results demonstrated that the depletion of NACA, but not NACB, was able to induce apoptosis. Deletion of middle or C-term rather than N-term induced obvious apoptosis. The middle part of NACA was response to bind NACB and form a complex. Without middle part, NACA redistributed into nuclei. We conclude NACA against apoptosis is independent of β subunit. C-term of NACA, which is identified as ubiquitin binding domain, and may take important role in anti-apoptosis function. Keywords: NACA, B-cell lymphoma, anti-apoptosis, depletion mutant.Download Full Article |
Editor’s Choice : Molecular Genetic Study of the Allelic State of the Cell Cycle Genes (TP53, BRCA1) and Features of the Regulation of the Cytokine Cascade in Breast Cancer
|
|
Abstract: This article contains the analysis of mutations in genes that regulate the cell cycle (TP53 and BRCA1) and classification relating to tumor suppressor. Shown that the "risk" alleles of these genes may contribute to tumor development, but the activation of the immune system cytokine spectrum of patients can prevent their destructive degeneration. The authors proposed a personalized approach to the study for the prevention of possible proliferative processes. This is confirmed by reversal of "risk" alleles studied genes in tumors in operated patients with cytokine physiologically normal status. Keywords: Breast cancer, tumor suppressor, cytokines, nucleotide substitutions, predisposition.Download Full Article |
Editor’s Choice : Patterns of Distant Failure and Second Primary Cancers in Patients with Oropharyngeal Squamous Cell Carcinoma: Implications for Surveillance Methodology
|
|
Abstract: Background: We analyzed the pattern of distant metastasis (DM) and secondary primary cancers (SPC) in patients with oropharyngeal squamous cell carcinoma (OPSCC) to develop surveillance guidelines. Methods: A retrospective review of 177 patients with OPSCC treated with intensity modulated radiation therapy ± chemotherapy between 2002 and 2012 was performed to characterize the rate, pattern, and timing of DM and SPC. Results: Sixteen patients (9.0%) developed DM and 9 patients (5.1%) developed a SPC. Overall, 24/177 patients (13.6%) developed a DM and/or SPC for a total of 27 events. 92.6% (25/27) of events were detectable on physical exam and/or chest CT. p16+ patients developed DM later than p16- tumors (23.4 months versus 8.7 months). Conclusions: Chest CT with physical examination detects the majority of DM and SPC in patients with OPSCC and would provide effective surveillance in these patients. A risk adapted surveillance strategy is proposed. Keywords: Oropharynx cancer, surveillance, survivorship care, distant metastasis, second primary cancer.Download Full Article |