The Expression of Homo Sapiens microRNA-21 (Hsa-miR-21-5p) and mRNA Reversion Inducing Cysteine Rich Protein with Kazal Motifs (RECK) in Plasma of Epithelial Ovarian Cancer

ABSTRACT Epithelial Ovarian Cancer (EOC) is malignant cancer that caused death for most women in Indonesia. The emergence of EOC showed no specific symptoms in its early stages; that makes the screening mostly occur when patients are in advanced stage. Treatment of EOC at an advanced stage will be more challenging with poor prognosis. Therefore, minimally invasive biomarkers are needed to diagnose at the early stage. microRNA is one of the potential biomarkers which not only expressed inside the cell but also secreted outside the cell with exosome protection. This protection makes microRNA stable. Moreover, several studies have shown the ability to detect microRNA in the blood sample. microRNA-21 (miR-21) is oncomiR which targeted tumor suppressor mRNA RECK based on in silico analysis.The first aim is to determine the expression of miR-21 in plasma samples of EOC patients compared with healthy controls. The second aim is to investigate the expression correlation between miR-21 and RECK mRNA.Blood samples were collected from 30 patients and 30 healthy controls. Plasma was then obtained from centrifugated blood samples. The total RNA was isolated and reverse transcribed to produce cDNAs. cDNAs were then quantified using qPCR using specific primer for miR-21 and RECK mRNA. The expression analysis was done relative expression method by Livak. The expression of miR-21 was calculated using the miR-16 expression as the reference gene. Also, Beta-actin was used as reference gene for RECK mRNA calculation. The correlation between the expression of miR-21 and mRNA RECK was analyzed using the Spearman rho correlation analysis.In this study showed the expression of miR-21 in patients with EOC increased 4.7579-fold compared with healthy controls (p <0.05). On the other hand, the miR-21 target, RECK mRNA, decreased 4,2 times with fold change 0.237665 on plasma EOC patients compared with healthy controls (p <0.05). The statistical calculation of the expression of miR-21 and mRNA RECK was inversely proportional to mRNA RECK with a strong correlationThis study has been able to prove that the expression of miR-21 is up-regulated in EOC patients and confirmed by the down-regulation of RECK expression. The next research challenge is to make anti-miR-21 to suppress the expression of miR-21 in EOC, which can be analyzed the effect of miR-21 in the development of EOC

Primary treatment results of Nasopharyngeal Carcinoma (NPC) in Yogyakarta, Indonesia

INTRODUCTION Nasopharyngeal Carcinoma (NPC) is a major health problem in southern and eastern Asia. In Indonesia NPC is the most frequent cancer in the head and neck area. NPC is very sensitive to radiotherapy resulting in 3-year disease-free and overall survival of approximately 70% and 80%, respectively. Here we present routine treatment results in a prospective study on NPC in a top referral; university hospital in Indonesia. METHODS All NPC patients presenting from September 2008 till January 2011 at the ear, nose and throat (ENT) department of the Dr. Sardjito General Hospital, Universitas Gadjah Mada, Yogyakarta, Indonesia, were possible candidates. Patients were included if the biopsy was a histological proven NPC without distant metastasis and were assessed during counselling sessions prior to treatment, as being able to complete the entire treatment. RESULTS In total 78 patients were included for treatment analysis. The median time between diagnosis and start of radiotherapy is 120 days. Forty-eight (62%) patients eventually finished all fractions of radiotherapy. The median duration of the radiotherapy is 62 days for 66 Gy. Median overall survival is 21 months (95% CI 18–35) from day of diagnosis. CONCLUSION The results presented here reveal that currently the treatment of NPC at an Indonesian hospital is not sufficient and cannot be compared to the treatment results in literature. Main reasons for these poor treatment results are (1) a long waiting time prior to the start of radiotherapy, (2) the extended overall duration of radiotherapy and (3) the advanced stage of disease at presentation.Maarten A. Wildeman, Renske Fles, Camelia Herdini, Rai S. Indrasari, Andrew D. Vincent, Maesadji Tjokronagoro, Sharon Stoker, Johan Kurnianda, Baris Karakullukcu, Kartika W. Taroeno- Hariadi, Olga Hamming-Vrieze, Jaap M. Middeldorp, Bambang Hariwiyanto, Sofia M. Haryana, I. Bing Ta

Biological activity, quantitative structure&ndash;activity relationship analysis, and molecular docking of xanthone derivatives as anticancer drugs

Isnatin Miladiyah,1,2 Jumina Jumina,3 Sofia Mubarika Haryana,4 Mustofa Mustofa5 1Pharmacology Department, Faculty of Medicine, Islamic University of Indonesia, 2Doctorate Program of Medical Science and Health, Faculty of Medicine, 3Chemistry Department, Faculty of Mathematics and Natural Sciences, 4Histology and Cell Biology Department, Faculty of Medicine, 5Pharmacology and Therapeutic Department, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia Background: Xanthone derivatives have a wide range of pharmacological activities, such as those involving antibacterial, antiviral, antimalarial, anthelmintic, anti-inflammatory, antiprotozoal, and anticancer properties. Among these, we investigated the anticancer properties of xanthone. This research aimed to analyze the biological activity of ten novel xanthone derivatives, to investigate the most contributing-descriptors for their cytotoxic activities, and to examine the possible mechanism of actions of xanthone compound through molecular docking. Materials and methods: The cytotoxic tests were carried out on WiDR and Vero cell lines, by a 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay method. The structural features required for xanthone&rsquo;s anticancer activity were conducted by using the semi-empirical Austin Model-1 method, and continued with quantitative structure-activity relationship (QSAR) analysis using BuildQSAR program. The study of the possible mechanism of actions of the selected xanthone compound was done through molecular docking with PLANTS. Results: The three novel xanthone derivatives (compounds 5, 7, and 8) exhibited cytotoxic activity with compound 5 showed the highest degree of cytotoxicity at concentration 9.23 &micro;g/mL (37.8 &micro;M). The following best equation model was obtained from the BuildQSAR calculation: log 1/IC50 = -8.124 qC1 -35.088 qC2 -6.008 qC3 + 1.831 u + 0.540 logP -9.115 (n = 10, r = 0.976, s = 0.144, F = 15.920, Q2 = 0.651, SPRESS = 0.390). This equation model generated 15 proposed new xanthone compounds with better-predicted anticancer activities. A molecular docking study of compound 5 showed that xanthone formed binding interactions with some receptors involved in cancer pathology, including telomerase, tumor-promoting inflammation (COX-2), and cyclin-dependent kinase-2 (CDK2) inhibitor. Conclusion: The results suggested that compound 5 showed the best cytotoxic activity among the xanthone derivatives tested. QSAR analysis showed that the descriptors contributed to xanthone&rsquo;s cytotoxic activity were the net atomic charge at qC1, qC2, and qC3 positions, also dipole moment and logP. Compound 5 was suspected to be cytotoxic by its inhibition of telomerase, COX-2, and CDK2 receptors. Keywords: xanthones, anticancer, semiempirical AM1, BuildQSAR, molecular dockin

Molecular diversity of Epstein-Barr virus IgG and IgA antibody responses in nasopharyngeal carcinoma: a comparison of Indonesian, Chinese, and European subjects.

Epstein-Barr virus (EBV)-specific immunoblot analysis was used to reveal the molecular diversity of immunoglobulin (Ig) G and IgA antibody responses against Epstein-Barr nuclear antigen (EBNA), early antigen (EA), and viral capsid antigen (VCA) in serum samples from patients with nasopharyngeal carcinoma (NPC) and control subjects, by use of immunofluorescence assay (IFA). Control donors (n=150) showed IgG responses to few EBV proteins--VCA-p18, VCA-p40, EBNA1, and Zebra--and sporadically weak IgA reactivity to EBNA1 and VCA-p18. Patients with NPC stage 1 (n=6) had similar response patterns. Patients with NPC stage 2-4 (n=132) showed significantly more diverse IgG and IgA responses to EA and VCA proteins--VCA-p18/-p40, EBNA1, Z-encoded broadly reactive activator, and EAd-p47/54, -DNAse, -thymidine kinase, and -p138. No correlation was found between IFA titers and the number of EBV proteins recognized by IgG or IgA. Our results reveal dissimilarity between EBV polypeptides recognized by IgG and IgA antibodies, which suggests independent B cell triggering events

Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia

Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (n = 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (P < 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = -0.91; P < 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited