Identification of commonly regulated protein targets and molecular pathways in PC-3 and DU145 androgen-independent human prostate cancer cells treated with the curcumin analogue 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one

Objective: To identify mutually regulated proteins in PC-3 and DU145 androgen-independent prostate cancer cell lines treated with 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one(MS17), and to study the molecular pathways that contributed to the anticancer activity of MS17.Methods: PC-3 and DU145 cells were treated with 3 × EC_(50)(15 μM) concentration of MS17 for 24 h and were subjected to protein expression profiling using two-dimensional gel electrophoresis and protein identification by mass spectrometry.Selected differentially expressed proteins with significant P-value of P<0.05 and fold change over 1.5-folds were filtered through and ontologically classified.Mutually regulated proteins were ranked by fold change and identified as common protein targets of MS17.Results: Profiling data revealed that, the mutually down-regulated proteins included ACTB and ACTG associated with structural molecule activity, ACTN1 with cell cycle, ACTN4 with cell migration, HNRPK with apoptosis, PLST with morphogenesis and TERA with proteolysis.However, the expressions of CH60 and HS71 A respectively associated with response to unfolded protein demonstrated opposing regulation in PC-3 and DU145 cells.Pathway analysis of the differentially expressed proteins in PC-3 cells demonstrated the modulation of top pathways associated with cell-cell adhesion and cytoskeletal organization while in DU145 cells the pathways were associated with proteosomal degradation, regulation of electrolytes and water, regulation control of germ cells and organization of filament assembly/disassembly.Conclusions: The findings of the present study provide an understanding on the anti-tumorigenic activity of MS17 at the proteome level and warrant further research for its potential application for the management and treatment of androgen-independent prostate cancer.

Identification of commonly regulated genes in HPV18-and HPV16-infected cervical cancer cells treated with the curcumin analogue 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one

Objective: To identify commonly regulated genes in HPV-infected He La and Ca Ski cervical cancer cells treated with curcumin analogue 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one(MS17) and to explore potential mechanisms that underlie its cytotoxic, anti-proliferative and apoptotic activity. Methods: He La and Ca Ski cells were treated with 2×EC50 and 3×EC50 doses of MS17 for 24 h and the RNA extracts were subjected to one-colour microarray-based gene expression profiling. Pair-wise significant genes(false discovery rate-corrected, P<0.05) were analysed for fold change(FC) compared to control samples. Differentially expressed genes with FC≥2.0(up-regulated; FC≥2.0 and down-regulated; FC≤-2.0) compared to the control samples were filtered through and analysed to create a global gene expression profile. Mutually regulated genes were ranked by FC and categorised by gene ontology. Results: Our data indicated dose-dependent regulation by MS17 and identified top 20 mutually upand down-regulated genes each in He La and Ca Ski cells. Amongst these 17 were commonly regulated in both cell lines. These include the up-regulation of CCL26, DEFB103 B, IL1 RL1, LY96, GCNT3, MMP10, MMP3, GADD45 G and HSPA6, and the down-regulation of TENM2, NEBL, KIFC1, CTDSP1, IGFBP5, LTBP1, NREP and MXD3. These genes were associated with key biological functions that were proposed to mediate the anticancer activity of MS17 in cervical cancer cells such as immune response, metabolic processes, proteolysis, programmed cell death, unfolded protein response, cell adhesion, cytoskeletal organisation, phosphatase activity, signal transduction and transcription regulator activity. Conclusions: Identification of seventeen common genes modulated by MS17 could be used as potential therapeutic targets in both cervical cancer cell lines and the findings of this study could be used to present an insight into the potential antitumor activity of MS17 in cervical cancer.

A community-based prospective cohort study of dengue viral infection in Malaysia: the study protocol

Background:Globally,dengue infections constitute a significant public health burden.In recent decades,Malaysia has become a dengue hyper-endemic country with the co-circulation of the four dengue virus serotypes.The cyclical dominance of sub-types contributes to a pattern of major outbreaks.The consequences can be observed in the rising incidence of reported dengue cases and dengue related deaths.Understanding the complex interaction of the dengue virus,its human hosts and the mosquito vectors at the community level may help develop strategies for addressing the problem.Methods:A prospective cohort study will be conducted in Segamat district of Johor State in Peninsular Malaysia.Researchers received approval from the Malaysian Medical Research Ethics Committee and Monash University Human Research Ethics Committee.The study will be conducted at a Malaysian based health and demographic surveillance site over a 1 year period in three different settings(urban,semi-urban and rural).The study will recruit healthy adults(male and female)aged 18 years and over,from three ethnic groups(Malay,Chinese and Indian).The sample size calculated using the Fleiss method with continuity correction is 333.Sero-surveillance of participants will be undertaken to identify asymptomatic,otherwise healthy cases;cases with dengue fever who are managed as out-patients;and cases with dengue fever admitted to a hospital.A genetic analysis of the participants will be undertaken to determine whether there is a relationship between genetic predisposition and disease severity.A detailed medical history,past history of dengue infection,vaccination history against other flaviviruses such as Japanese encephalitis and Yellow fever,and the family history of dengue infection will also be collected.In addition,a mosquito surveillance will be carried out simultaneously in recruitment areas to determine the molecular taxonomy of circulating vectors.Discussion:The research findings will estimate the burden of asymptomatic and symptomatic dengue at the community level.It will also examine the relationship between virus serotypes and host genotypes,and the association of the clinical manifestation of the early phase with the entire course of illness.

Seroprevalence of dengue among healthy adults in a rural community in Southern Malaysia:a pilot study

Background:The frequency and magnitude of dengue epidemics continue to increase exponentially in Malaysia,with a shift in the age range predominance toward adults and an expansion to rural areas.Despite this,information pertaining to the extent of transmission of dengue virus(DENV)in the rural community is lacking.This communitybased pilot study was conducted to establish DENV seroprevalence amongst healthy adults in a rural district in Southern Malaysia,and to identify influencing factors.Methods:In this study undertaken between April and May 2015,a total of 277 adult participants were recruited from households across three localities in the Sungai Segamat subdistrict in Segamat district.Sera were tested for immunoglobulin G(IgG)(Panbio®Dengue Indirect IgG ELISA/high-titer capture)and immunoglobulin M(IgM)(Panbio®)antibodies.The plaque reduction neutralization test(PRNT)was conducted on random samples of IgGpositive sera for further confirmation.Medical history and a recall of previous history of dengue were collected through interviews,whereas sociodemographic information was obtained from an existing database.Results:The overall seroprevalence for DENV infection was 86.6%(240/277)(95%CI:83-91%).Serological evidence of recent infection(IgM/high-titer capture IgG)was noted in 11.2%(31/277)of participants,whereas there was evidence of past infection in 75.5%(209/277)of participants(indirect IgG minus recent infections).The PRNT assay showed that the detected antibodies were indeed specific to DENV.The multivariate analysis showed that the older age group was significantly associated with past DENV infections.Seropositivity increased with age;48.5%in the age group of<25 years to more than 85%in age group of>45 years(P<0.001).No associations with occupation,study site,housing type,comorbidity,educational level,and marital status were observed,although the latter two were statistically significant in the univariate analysis.None of the studied factors were significantly associated with recent DENV infections in the multivariate analysis,although there was a pattern suggestive of recent outbreak in two study sites populated predominately by Chinese people.The majority of infections did not give rise to recognizable disease(either asymptomatic or nonspecific symptoms)as only 12.9%of participants(31/240)recalled having dengue in the past.Conclusions:The predominantly rural community under study had a very high previous exposure to dengue.The finding of a high proportion of unreported cases possibly due to subclinical infections underscores the need for enhanced surveillance and control methods.This finding also has implications for measuring disease burden,understanding transmission dynamics,and hypothesizing effects on DENV vaccine efficacy and uptake.