microRNA-21在食管癌细胞增殖、迁移和凋亡中的作用

本研究检测了40例食管癌组织和40例癌旁组织中的miR-21、PTEN、PI3K和AKT表达,并通过转染miR-21抑制剂来敲低人食管癌细胞系EC9706的miR-21表达,考察了miR-21对食管癌细胞生长的影响。研究发现,食管癌组织中PTEN蛋白的阳性染色评分低于癌旁组织(p<0.05),而PI3K和AKT蛋白的阳性染色评分高于癌旁组织(p<0.05)。miR-21在人食管癌组织中被上调(3.56 vs 1.21,p<0.05)。转染miR-21抑制剂导致PTEN蛋白表达升高,而PI3K和AKT蛋白表达降低(p<0.05)。转染miR-21抑制剂抑制了EC9706细胞的增殖和迁移,但促进了细胞凋亡(p<0.05)。miR-21的上调可通过激活PTEN/PI3K/AKT信号通路来促进食道癌细胞的增殖和迁移,并抑制细胞凋亡。     

羽扇豆醇通过抑制RhoA-ROCK1信号通路抑制结肠癌细胞增殖

该文探讨了羽扇豆醇(Lupeol)对人结肠癌HCT116和SW620细胞增殖的影响及相关作用机制。使用不同浓度的Lupeol处理HCT116和SW620细胞后,用MTT法检测细胞活性,CCK8法检测细胞增殖能力,平板克隆实验检测细胞克隆形成能力,流式细胞术检测细胞周期和细胞凋亡,(quantitative real-time PCR,qPCR)和Western blot检测相应mRNA和蛋白表达水平,免疫荧光检测β-Catenin蛋白细胞内分布情况。通过构建shRNA敲低两种结肠癌细胞中RhoA,进一步研究Lupeol影响细胞增殖的分子机制。结果显示,Lupeol处理后,HCT116和SW620细胞增殖能力明显下降,克隆形成能力受到抑制,细胞周期阻滞于G0/G1期,细胞内RhoA、ROCK1、β-Catenin、Cyclin D1 mRNA和蛋白表达水平均显著下降,β-Catenin蛋白胞质和胞膜上分布减少。敲低RhoA后抑制了细胞增殖,同时使得RhoA-ROCK1-β-Catenin信号通路蛋白受到抑制,β-Catenin蛋白胞质和胞膜上分布减少。综上所述,Lupeol可通过抑制RhoA-ROCK1信号通路,抑制β-Catenin蛋白表达,进而抑制HCT116和SW620细胞增殖,Lupeol有望成为临床结肠癌治疗的新药物。     

Improved therapeutic efficiency of photothermal treatment and nursing care in prostate cancer by DOX loaded PEG coated Cu@Se nano-hybrid vesicle

Cancer is one of the major life threatening diseases, with higher mortality rate and morbidity. It is always a challenge for effective drug delivery and release of drug in specific tumor sites. Therefore to identify the synergistic effect of chemotherapeutic drug and photo thermal agent on tumor area, Doxorubicin (DOX) acts as anticancer agent but it has low aqueous solubility so its clinical application is limited. The present study developed doxorubicin (DOX) were designed to be with the poly ethylene glycol (PEG) functionalized copper and selenium (Cu-Se) nanoparticles (PEG@Cu-Se+DOX) and it is efficiently synthesized and enhance its aqueous formulation and improve the prostate cancer (DU145 and LNCaP) activity. The characteristics like mono dispersity, size stability and constant spectral of as-synthesized nanoparticles are comparably excellent than DOX alone. Also the enhanced cellular uptake and in vitro cytoxicicty suggests these nanoparticles selectively killing prostate cancer. In this present study explained that PEG@Cu-Se+DOX as a safe and hopeful strategy for chemotherapeutics of photothermal therapy and deserve for further clinical evaluations.     

Next generation sequencing and functional pathway analysis to understand the mechanism of action of copper-tolfenamic acid against pancreatic cancer cells

Anti-cancer activity of tolfenamic acid (TA) in preclinical models for pancreatic cancer (PaCa) is well established. Since the dosage for anti-cancer actions of TA is rather high, we recently demonstrated that IC₅₀ values of Copper-TA are 30–80% less than TA in 12 cancer cell lines. This study elucidates the underlying mechanisms of Copper-TA in PaCa cells. Control and Copper-TA (IC₅₀) treated PaCa cells were processed by next-generation sequencing (NGS) to determine differentially expressed genes using HTG EdgeSeq Oncology Biomarker panel. Ingenuity Pathway Analysis (IPA®) was used to identify functional significance of altered genes. The conformational studies for assessing the expression of key regulators and genes were conducted by Western blot and qPCR. IPA® identified several networks, regulators, as well as molecular and cellular functions associated with cancer. The top 5 molecular and cellular functions affected by Cu-TA treatment were cell death and survival, cellular development, cell growth and proliferation, cell cycle and cellular movement. The expression of top upstream regulators was confirmed by Western blot analysis while qPCR results of selected genes demonstrated that Copper-TA is efficacious at lower doses than TA. Results suggest that Copper-TA alters genes/key regulators associated with cancer and potentially serve as an effective anti-cancer agent.     

Nontyphoidal Salmonella: a potential anticancer agent

Use of bacteria in cancer therapy, despite being considered as a potent strategy, has not really picked up the way other methods of cancer therapies have evolved. However, in recent years, the interest on use of bacteria to kill cancer cells has renewed considerably. The standard and widely followed strategies of cancer treatment often fail either due to the complexity of tumour biology or because of the accompanying side effects. In contrast, these limitations can be easily overcome in a bacteria-mediated approach. Salmonella is a bacterium, which is known for its ability to colonize solid or semisolid tumours more efficiently than any other bacteria. Among more than 2500 serovars of Salmonella, S. Typhimurium has been widely studied for its antagonistic effects on cancer cells. Here in, we review the current status of the preclinical and the clinical studies with a focus on the mechanisms that attribute the anticancer properties to nontyphoidal Salmonella.     

Choosing between conventional and hypofractionated prostate cancer radiation therapy: Results from a study of shared decision-making

AimTo evaluate patient choice of prostate cancer radiotherapy fractionation, using a decision aid.BackgroundRecent ASTRO guidelines recommend patients with localised prostate cancer be offered moderately hypofractionated radiation therapy after discussing increased acute toxicity and uncertainty of long-term results compared to conventional fractionation.Materials and methodsA decision aid was designed to outline the benefits and potential downsides of conventionally and moderately hypofractionated radiation therapy. The aid incorporated the ASTRO guideline to outline risks and benefits.ResultsIn all, 124 patients with localised prostate cancer were seen from June-December 2018. Median age was 72 (range 50–90), 49.6 % were intermediate risk (50.4 % high risk). All except three patients made a choice using the aid; the three undecided patients were hypofractionated. In all, 33.9 % of patients chose hypofractionation: falling to 25.3 % for patients under 75 years, 24.3 % for patients living within 30 miles of the cancer centre, and 14.3 % for patients with baseline gastrointestinal symptoms. On multivariate analysis, younger age, proximity to the centre, and having baseline gastrointestinal symptoms significantly predicted for choosing conventional fractionation. Insurance status, attending clinician, baseline genitourinary symptoms, work/carer status, ECOG, cancer risk group and driving status did not impact choice. Reasons for choosing conventional fractionation were certainty of long-term results (84 %) and lower acute bowel toxicity (51 %).ConclusionsMost patients declined the convenience of moderate hypofractionation due to potentially increased acute toxicity, and the uncertainty of long-term outcomes. We advocate that no patient should be offered hypofractionation without a thorough discussion of uncertainty and acute toxicity.     

Evaluating the predictive value of quantec rectum tolerance dose suggestions on acute rectal toxicity in prostate carcinoma patients treated with IMRT

AimTo investigate the predictive value of convenience of rectum dosimetry with Quantitative Analysis of Normal Tissue Effects in the Clinic (QUANTEC) dose limits, maximum rectum dose (Dmax), total rectal volume (TVrectum), rectal volume included in PTV (VrectumPTV) on Grade 2–3 acute rectal toxicity for utilization in clinical practice.BackgroundNumerous previous data have reported frequent acute proctitis after external-beam RT of prostate cancer. Predicting toxicity limited with dose information is inadequate in clinical practice due to comorbidities and medications used.Materials and MethodSixty-four non-metastatic prostate cancer patients treated with IMRT were enrolled. Patients were treated to a total dose of 70–76 Gy. Rectal dose volume histograms (DVH) of all patients were evaluated retrospectively, and a QUANTEC Score between 0 and 5 was calculated for each patient. The correlation between the rectal DVH data, QUANTEC score, TVrectum, VrectumPTV, rectum Dmax and Grade 2–3 rectal toxicity was investigated.ResultsIn the whole group grade 1, 2 and 3 acute rectal toxicities were 25%, 18.8% and 3.1%, respectively. In the DVH data, rectum doses of all patients were under RTOG dose limits. Statistically significant correlation was found between grade 2–3 rectal toxicity and TVrectum (p = 0,043); however. It was not correlated with QUANTEC score, VrectumPTV and Dmax.ConclusionOur results were not able to show any significant correlation between increasing convenience with QUANTEC limits and lower rectal toxicity. Conclusively, new dosimetric definitions are warranted to predict acute rectal toxicity more accurately in prostate cancer patients during IMRT treatment.     

Surgery versus radiotherapy: Long term outcomes of T1 glottic cancer

AimThe aim of this study was to compare the outcomes, patterns of failure and laryngeal preservation rates in patients with T1N0 glottic cancer treated with surgery or radiotherapy.Materials/methodsRetrospective study of T1N0 glottic cancer patients treated in our institution between January 2007 and December 2017. Histologically proven squamous cell carcinoma patients, treated with upfront cordectomy/partial laryngectomy (S group) or radiotherapy (RT group) were included. Elective treatment of the neck was not permitted. Local failure (LF), disease-free survival (DFS), ultimate disease-free survival (UDFS), laryngectomy-free survival (LFS), disease-specific mortality (DSM) and overall survival (OS) were evaluated.ResultsTwo hundred and one patients were eligible (172 S group, 29 RT group), with a median follow-up of 38.8 months. Overall, 33 (16%) patients had a recurrence, 30 (17%) in the S group and 3 (10%) in the RT group. Local failure was the predominant site of failure (28 S, 2 RT). Overall, of all those that were salvaged, 17 (8%) underwent total laryngectomy (15 S, 2 RT). There was no significant difference in the 5-year cumulative incidence of LF (20.8% S, 8.1% RT, p = 0.138), 5-y LFS (85.0% vs. 91.7%, p = 0.809), 5-y DFS (67.5% vs. 82.1%, p = 0.343), 5-y UDFS (82.5% vs. 90.3%, p = 0.647) and 5-y OS (84.5% vs. 90.3%, p = 0.892). Multivariate analysis showed no correlation between initial treatment and the analyzed outcomes.ConclusionPrimary surgery or radiotherapy were similar first line options, since they do not differ in all outcomes. Patients’ and physician's preferences must be considered when choosing first treatment.     

Reporter mice for isolating and auditing cell type‐specific extracellular vesicles in vivo

Extracellular vesicles (EVs) are abundant, lipid‐enclosed vectors that contain nucleic acids and proteins, they can be secreted from donor cells and freely circulate, and they can be engulfed by recipient cells thus enabling systemic communication between heterotypic cell types. However, genetic tools for labeling, isolating, and auditing cell type‐specific EVs in vivo, without prior in vitro manipulation, are lacking. We have used CRISPR‐Cas9‐mediated genome editing to generate mice bearing a CD63‐emGFP^{loxP/stop/loxP} knock‐in cassette that enables the specific labeling of circulating CD63⁺ vesicles from any cell type when crossed with lineage‐specific Cre recombinase driver mice. As proof‐of‐principle, we have crossed these mice with Cdh5‐Cre^ERT2 mice to generate CD63^emGFP+ vasculature. Using these mice, we show that developing vasculature is marked with emerald GFP (emGFP) following tamoxifen administration to pregnant females. In adult mice, quiescent vasculature and angiogenic vasculature (in tumors) is also marked with emGFP. Moreover, whole plasma‐purified EVs contain a subpopulation of emGFP⁺ vesicles that are derived from the endothelium, co‐express additional EV (e.g., CD9 and CD81) and endothelial cell (e.g., CD105) markers, and they harbor specific miRNAs (e.g., miR‐126, miR‐30c, and miR‐125b). This new mouse strain should be a useful genetic tool for generating cell type‐specific, CD63⁺ EVs that freely circulate in serum and can subsequently be isolated and characterized using standard methodologies.