Regulation of HSP27 on NF-κB pathway activation may be involved in metastatic hepatocellular carcinoma cells apoptosis

<p>Abstract</p> <p>Background</p> <p>During the process of metastasis, cells are subjected to various apoptotic stimuli. Aberrant expression of apoptotic regulators often contribute to cell metastasis. Heat shock protein 27(HSP27) is confirmed as an apoptosis regulator, but its antiapoptotic mechanism in metastatic hepatocellular carcinoma (HCC) cells remains unclear.</p> <p>Methods</p> <p>Levels of HSP27 protein and its phosphorylation in Hep3B, MHCC97L to MHCC97H cells with different metastatic potentials were determined by western blot analysis. MHCC97H cells were transfected with specific small interference RNA (siRNA) against HSP27. The <it>in vitro </it>migration and invasion potentials of cells were evaluated by Transwell assay. The apoptosis ratio of MHCC97H cells was analyzed by TUNEL staining and Flow Cytometry. Alteration of signal transduction pathway after HSP27 knockdown in MHCC97H cells was evaluated through a Human Q Series Signal Transduction in Cancer Gene Array analysis. Nuclear NF-κB contentration and endogenous IKK activity were demonstrated by ELISA assay. The association of IKKα, IKKβ, IκBα with HSP27 and the association between IKKβ and IKKα in MHCC97H cells were determined by co-immunoprecipitation assay followed by western blot analysis.</p> <p>Results</p> <p>HSP27 protein and its phosphorylation increased in parallel with enhanced metastatic potentials of HCC cells. siRNA-mediated HSP27 knockdown in MHCC97H significantly suppressed cells migration and invasion <it>in vitro </it>and induced cell apoptosis; the prominently altered signal transduction pathway was NF-κB pathway after HSP27 knockdown in MHCC97H cells. Furthermore, inhibition of HSP27 expression led to a significant decrease of nuclear NF-κB contentration and endogenous IKK activity. In addition, HSP27 was associated with IKKα, IKKβ, IκBα in three HCC cells above. ELISA assay and western blot analysis also showed a decrease of the association between IKKβ and IKKα, the association between phosphor-HSP27 and IKK complex, and an increase of total IκBα but reducing tendency of phosphor-IκBα when HSP27 expression was efficiently knocked down in MHCC97H cells.</p> <p>Conclusion</p> <p>Altogether, these findings revealed a possible effect of HSP27 on apoptosis in metastatic HCC cells, in which HSP27 may regulate NF-kB pathway activation.</p

RNA biomarkers in colorectal cancer

Colorectal cancer (CRC) develops and progresses through a systematic selection for (epi) genetic alterations that drive the transformation from normal colon epithelium to adenocarcinoma. These changes affect both noncoding RNAs and mRNAs and so define the clinical behaviour of cancer cells within a distinctive host genetic and environmental context. Although earlier diagnosis and more effective treatment modalities have decreased mortality from CRC, prognostic stratification and adjuvant therapy selection after surgery remain dependent on broad descriptive classifications, opportune histological markers of poor prognosis and chemotherapy efficacy data derived from diverse CRC populations. Crucially, there is significant inter- and intra-individual variability in response to, and tolerance of, chemotherapy treatments. These limitations explain the small clinical benefit of new agents studied in contemporary phase III trials. Molecular assays have the potential to address these constraints and there has been intense interest in the identification of clinically relevant molecular biomarkers. These must be easy to obtain and quantify and ideally represent steps in well-understood carcinogenic pathways or host-response mechanisms. Although some biomarkers can provide broad prognostic information based on CRC subtype (e.g. MSI status) or can somewhat predict response to targeted therapies (e.g. KRAS), no RNA-based biomarkers have entered routine clinical practice. This is due, in part, to the genetic heterogeneity of both patients and CRC. In addition, serious underlying issues with regards to study design, poor technical protocols, inadequate quality controls and inappropriate data analysis prevent successful translation of research results. Consequently, the identification of clinically relevant panels of biomarkers will depend not just on further advances in our understanding of CRC biology, but will need to be coupled with appropriate study designs and more suitable, standardised and transparent techniques

Hitting the greens: Backcrossing the csn-5 mutant, par for the course, a CSN-5 protein null [abstract]

Abstract only availableGermline development is crucial to all eukaryotic organisms and the survival of their species. Dr. Bennett's laboratory utilizes Caenorhabditis elegans, the free-living soil worm and the Nobel-prize winning model organism, to study cytoplasmic granules called P granules. P granules contain both protein and RNA and are specifically associated with the germ cells. The germline RNA-helicase proteins (GLHs) are necessary components of the P granules; they are essential for fertility in the nematode. Several protein interactors with the GLHs were previously identified in a yeast-two hybrid screen. One of these GLH binding partners is CSN-5, a known component of the COP 9 signalosome, a complex involved in protein stability that is conserved from plants to worms to humans. Recently, a deletion mutant strain, csn-5(vc861), has been isolated by the C. elegans Knockout Consortium. My project involves backcrossing (also called outcrossing) the green GFP balanced csn-5(vc861) strain against wild-type worms to remove other possible mutations arising from the original mutagenesis. The other objective is to determine whether the homozygous non-green mutant is a CSN-5 protein null. To date, four backcrosses are finished with two to be completed before initiating crosses with other pertinent mutant strains. In addition, immunocytochemistry and western blot analyses were done using an anti-CSN-5 antibody we generated. Both of these assays show that csn-5(vc861) does not make any CSN-5 protein and therefore the strain is likely a null.Life Sciences Undergraduate Research Opportunity Progra

Active surveillance-much safety, little recruitment Is it possible to extend the indication for intermediate-risk prostate cancer?

Background In contrast to North America or Sweden, active surveillance (AS) has not yet become established in our country for suitable prostate carcinomas (PCa). The strict entry criteria specified by the guideline are not likely to improve the acceptance in the near future. In early detection, prostate-specific antigen (PSA) testing leads to high numbers of overtreatment. There are various reasons for the continued preference for radical surgery. Objectives The goal is to examine whether the heterogeneous group with intermediate-risk PCa contains tumors that may be eligible for AS. Materials and methods In the HAROW trial, 52 AS patients with differently defined intermediate-risk PCa were followed for a median of 85.6 months. Oncologic outcomes are reported. Results Sixteen (30%) patients had a tumor of cT2b category, 21 (40%) had a Gleason score 3 + 4, 7 (14%) had >= 3 positive biopsy cores, 21 (40%) had a PSA >10 ng/ml, and 22 (42%) had a PSA density >0.2 ng/ml(2). Carcinoma-specific and metastasis-free survival were 100% and 96%, respectively. Thirty four patients discontinued AS in favor of invasive treatment, and an additional eight men maintained a noninvasive approach by switching to watchful waiting. Conclusions Efforts are under way to specify the criteria for patients with intermediate-risk PCa who may be eligible for AS. Tumors of cT2 category could be grouped together. The Gleason 4 fraction needs to be quantified because it determines the prognosis

Noninvasive treatment of organ-confined prostate cancer in elderly patients-results of the HAROW study

Background Noninvasive treatment options such as active surveillance (AS), watchful waiting (WW), and hormone deprivation therapy (HT) are particularly important in elderly patients with localized prostate cancer (PCa). Objectives We examine the use of these noninvasive treatment options in the everyday care in a cohort of patients >= 70 years old. Materials and methods In the HAROW study, the treatment of localized PCa under everyday conditions is investigated. The only inclusion criterion was newly diagnosed organ-confined PCa (= 70 years, 210 chose AS, 160 HT, and 87 WW. Observation times were 6.3x202f;years (AS), 7.5x202f;years (HT), and 7.0x202f;years (WW). AS patients (73.2x202f;years) were younger than WW (76.0x202f;years) and HT patients (76.9x202f;years) and had a higher proportion of low-risk tumors (80%) versus WW (31%) and HT (19%). A change of therapy was observed in 47.1% of AS, 17.2% of WW and 13.1% of HT patients. Metastasis occurred in 1.0% of AS, 4.6% of WW, and 6.9% of HT patients. Overall survival was 94.3% for AS, 90.8% for WW and 81.9% for HT. Within the first 28.4 months, the mean number of PSA determinations did not differ between AS and WW (6.1 vs. 5.2; px202f;= 0.09); a rebiopsy was performed in 37.6% of AS, 11.4% of WW, and 17% of HT patients. Conclusions The allocation to curative and palliative strategies should be made according to patient and tumor characteristics by definition. Palliative procedures may represent concepts in older patients who initially chose a curative AS strategy

Active Surveillance for Incidental (cT1a/b) Prostate Cancer: Long-Term Outcomes of the Prospective Noninterventional HAROW Study

Introduction: Optimal treatment for incidental prostate cancer (IPC) after surgical treatment for benign prostate obstruction is still debatable. We report on long-term outcomes of IPC patients managed with active surveillance (AS) in a German multicenter study. Methods: HAROW (2008-2013) was designed as a noninterventional, prospective, health-service research study for patients with localized prostate cancer (= 0.2 ng/mL(2) (RR: 13.23; p = 1.0 ng/mL after surgery (RR: 5.19; p = 0.016) were significantly predictive for receiving an invasive treatment. Conclusion: In comparison with other AS series with a general low-risk prostate cancer population, our study confirmed the promising survival outcomes for IPC patients, whereas discontinuation rates seem to be lower for IPC. Thus, IPC patients at low risk of progression may be good candidates for AS