Glypican-3 mRNA expression level in Wilms tumor: correlation with histological type,stage, and outcome

Purpose

To correlate expression of Glypican-3 in Wilms tumor with histopathology, stage, and outcome.

Methods

Glypican-3 mRNA expression by real-time PCR on tumor and normal germline samples from 75 fresh nephrectomies for Wilms tumor with fold change after normalization against GAPDH was compared. Survival analysis for event-free and overall survival (EFS, OS) with 2-year follow-up for Glypican-3 overexpression (>1.5 times) and clinicopathological parameters was performed.

Results

Glypican-3 was overexpressed in 37/75 (49.3%). It was overexpressed in 77% (10/13) cases with blastema predominance or anaplastic histology, as compared to 44% of other histologies (27/62) (p = 0.03). OS was 73 and 93%, respectively (p = 0.016), for those with and without GPC-3 overexpression. EFS was not significantly different with Glypican-3 overexpression (p = 0.11). All 5 deaths among blastema predominant tumors and 4/5 deaths among triphasic tumors had overexpressed Glypican-3. Most deaths in Stage IV, Stage III, and Stage I + II (5/7, 3/3, 1/1) had GPC-3 overexpression. On multivariate analysis, only histology and stage were found to have independent prognostic value.

Conclusion

Glypican-3 overexpression in Wilms tumor correlates with poor OS on univariate analysis. However, only histology and stage have independent prognostic value. Glypican-3 levels may help to stratify intermediate outcome histology (triphasic) and Stage III Wilms tumors.

     

Ethyl acetate extract of Nepenthes adrianii x clipeata induces antiproliferation,apoptosis, and DNA damage against oral cancer cells through oxidative stress

Nepenthes plants are regarded as a kind of Traditional Chinese Medicine for several diseases but its anticancer activity remain unclear. The subject of this study is to evaluate the antiproliferation effects on oral cancer cells by Nepenthes plants using ethyl acetate extract of Nepenthes adrianii x clipeata (EANA). Cell viability was detected using MTS assay. Its detailed mechanisms including cell cycle, apoptosis, oxidative stress, and DNA damage were explored by flow cytometry or western blotting. For 24 hours EANA treatment, five kinds of oral cancer cells (CAL 27, Ca9‐22, OECM‐1, HSC‐3, and SCC9) show IC₅₀ values of cell viability ranging from 8 to 17 μg/mL but the viability of normal oral cells (HGF‐1) remains over 80%. Subsequently, CAL 27 and Ca9‐22 cells with high sensitivity to EANA were chosen to investigate the detailed mechanism. EANA displays the time course and concentration effects for inducing apoptosis based on flow cytometry (subG1 and annexin V analyses) and western blotting [cleaved poly (ADP‐ribose) polymerase (c‐PARP)]. Oxidative stress and DNA damage were induced by EANA treatments in oral cancer cells through reactive oxygen species (ROS), mitochondrial membrane potential disruption, mitochondrial superoxide, and γH2AX. All these changes of EANA treatments in oral cancer cells were reverted by the ROS scavenger N‐acetylcysteine pretreatment. Therefore, EANA induces preferential killing, apoptosis, and DNA damage against oral cancer cells through oxidative stress.     

MicroRNA-15a expression measured in urine samples as a potential biomarker of renal cell carcinoma

Introduction

Currently, there is no accurate diagnostic molecular biomarker for renal cell carcinoma (RCC). The aim of this study was to assess the expression of microRNA-15a (miR-15a) in urine of patients with RCC and to evaluate its potential as a diagnostic molecular biomarker.

Materials and methods

In total, 67 patients with solid renal tumors were enrolled: clear-cell RCC (ccRCC, n?=?22), papillary RCC (pRCC, n?=?16), chromophobe RCC (chRCC, n?=?14), oncocytoma (n?=?8), papillary adenoma (n?=?2) and angiomyolipoma (n?=?5). MiRNA-15a expression levels measurement in urine were performed using qPCR. Urine of 15 healthy volunteers without kidney pathology was used as control.

Results

We observed a difference in mean miR-15a expression values in groups of pre-operative patients with RCC, benign renal tumors and healthy persons (2.50E?01?±?2.72E?01 vs 1.32E?03?±?3.90E?03 vs 3.36E?07?±?1.04E?07 RFU, respectively, p?<?0.01). There was no difference in miR-15a expression between ccRCC, pRCC and chRCC (p?>?0.05). Direct association between RCC size and miR-15a expression values was obtained (Pearson correlation coefficient—0.873). On the 8th day after nephrectomy, mean expression value in patients with RCC decreased by 99.53% (p?<?0.01). MiR-15a expression differentiated RCC from benign renal tumors with 98.1% specificity, 100% sensitivity at a cut-off value of 5.00E?06 RFU, with AUC—0.955.

Conclusions

MiR-15a expression measured in urine may be used as diagnostic molecular biomarker for RCC.

     

Physical activity and sedentary behaviours of South Asian and white European children in inner city secondary schools in the UK

BACKGROUND: People of South Asian (SA) origin have an increased risk of premature coronary heart disease. In children of SA origin, there is an increased prevalence of obesity and evidence of insulin resistance. Risk factors for cardiovascular disease in children often persist into adulthood. Low levels of physical activity are likely to be linked to the rise in obesity. OBJECTIVE: To determine levels of physical activity and sedentary behaviours in secondary school pupils in the UK, including comparison of SA and white European (WE) children and those with and without a family history of cardiovascular disease. METHOD: Questionnaire survey conducted within an action research study in five inner city secondary schools serving a predominantly SA population. RESULTS: We obtained 3601 responses from 76% of eligible pupils. WE pupils were more likely to have walked to and from school compared to SAs. However, overall we identified low levels of physical activity and higher levels of inactive behaviours in both ethnic groups. Almost half (46%) of respondents spent four or more hours per day watching television or videos or playing computer games. An overall low level of active behaviour during school breaks was particularly emphasized in girls. We found no evidence of an association between physical activity levels and family history of cardiovascular disease. CONCLUSIONS: There is an urgent need for those with responsibility for young people's health, including parents, schools and community health providers, to consider and address the need for effective interventions to encourage increased physical activity levels.