The Indonesian Biomedical Journal

329-48

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Metastases are simply known as cancers spread to another part of the body, and often be responsible for the severity of cancer prognosis. Somehow, the complex mechanisms of metastases are not fully understood yet. CONTENT: The characteristic of cancer is akin to a never-healing wound. Cancer cells are plastic and dynamic as they build their niches and developed into metastases, even when they seem dormant. Therefore, cancer cells can survive the immune system. Recent research has shown the distinct biology of metastasis-initiating cell, which leads to tumor development in distant organs, immune surveillance evasion, and co-option of metastatic micro-environments. Effective cancer therapies must consider the regenerative states of metastatic malignancies and have careful observation of patient phenotypes. SUMMARY: This review aimed to provide an insight on genesis and characteristics of metastases, starting from its seeding and dormancy, until the advance phase. Thus, developing therapy for cancer metastases should not start as it grows, but even as earlier strategies since the primary tumor was detected.

358-64

Authors: Ferry Sandra, Muhammad Ihsan Rizal, Caecilia Caroline Aliwarga, Jenifer Christy Hadimartana, Maria Celinna

BACKGROUND: Caffeic acid has been reported to activate caspases in MG-63 osteosarcoma cells, which can lead to apoptosis via both extrinsic and intrinsic apoptotic pathways. Translocation of protein kinase C delta (PKCδ), which reduces mitochondrial membrane potential (ΔΨm), is involved in apoptosis. The role of PKCδ translocation and ΔΨm alteration in caffeic acid-induced MG-63 cell apoptosis are largely unknown. Present study investigated the effect of caffeic acid on PKCδ translocation and ΔΨm in MG-63 cells. METHODS: MG-63 cells were cultured and starved, followed by pretreatment with or without Z-VAD-FMK and treatment with or without 10 μg/mL caffeic acid. MG-63 cells were collected, lysed, and processed to obtain cytosolic and mitochondrial fractions. Each fraction was subjected to immunoblotting analysis by using anti-PKCδ antibody. Mitochondrial membrane potential (ΔΨm) was measured using flow cytometry. RESULTS: Cytosolic PKCδ levels were higher than mitochondrial PKCδ levels in untreated and 1 h caffeic acid treatment groups. Inversely, cytosolic PKCδ levels were lower than the mitochondrial PKCδ levels after 6 and 12 h caffeic acid treatment. By Z-VAD-FMK pretreatment, cytosolic PKCδ levels were higher than mitochondrial PKCδ after 6 and 12 h caffeic acid treatment. After 6 h treatment with caffeic acid, ΔΨm was slightly shifted. More shifting occurred in MG-63 cells treated with caffeic acid for 12 h. The ΔΨm shifting was inhibited by Z-VAD-FMK pretreatment. CONCLUSION: Caffeic acid could trigger apoptosis of MG-63 osteosarcoma cells by inducing PKCδ translocation to mitochondria and reducing ΔΨm, which might cause MMP.

376-81

Authors: Annisa Annisa, Elizabeth Henny Herningtyas, Dewajani Purnomosari

BACKGROUND: Hypoxia adaptation in cancer cells is mediated by hypoxia inducible factor (HIF)-1 that induces the expression of various proteins. Up to date, the analysis of T47D breast cancer cells-secreted protein is still limited. The aim of this study was to compare the protein profile secreted by T47D breast cancer cells under hypoxic and to normoxic conditions and identify the proteins as candidate for hypoxia marker proteins in T47D breast cancer cells. METHODS: T47D breast cancer cells were cultured under standard conditions. Cells were subcultured in normoxic and hypoxic conditions. The normoxic group was incubated with 20% oxygen and the hypoxic groups were incubated in a hypoxic chamber with 0.5% and 5% oxygen for 6, 24, and 48 hours in serum free medium. Proteins in the culture media were isolated and precipitated. Protein concentrations released by the cells were then measured by bicinchoninic assay (BCA). Protein bands were visualized by the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method. The bands that had differences between the hypoxic and normoxic groups were further analyzed by liquid chromatography-mass spectrophotometer (LC-MS). RESULTS: Hypoxic groups showed higher secretory protein than normoxic group. Protein bands were found in the 0.5% hypoxic group with a size of 50-75 kDa. Secretory proteins identified by LC-MS were keratin 1, 2, 9, and 10. CONCLUSION: The T47D cell line under 0.5% hypoxic treatment showed higher secretory proteins that identified as keratin 1, 2, 9, and 10.

393-400

Authors: Iskandar Rahardjo Budianto, Agus Firmansyah, Yefta Moenadjat, Ahmad Aulia Jusuf, Vivian Soetikno

BACKGROUND: Many hypotheses regarding the pathophysiology of enterocolitis in aganglionic megacolon or Hirschsprung disease (HSCR) has been proposed. Paneth cells are columnar intestinal epithelial cells that have an important role in maintaining of intestinal homeostasis as a bactericide. Since enterocolitis in HSCR may have association with Paneth cells metaplasia and hyperplasia, current study investigated Paneth cells metaplasia and hyperplasia in the sigmoid colon of HSCR rat model and its products, namely a-defensins and IL-1b, in the sigmoid colon tissues. METHODS: Aganglionosis-induced and control Sprague-Dawley rats were euthanized on Day (D)-7, -14, -17, -19, -21, -23, -25, and -28. Sigmoid colon tissue was isolated at each time point, and degree of enterocolitis as well as Paneth cells metaplasia and hyperplasia were analyzed by Hematoxylin-eosin staining, then protein levels of a-defensins and interleukin (IL)-1b were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Enterocolitis scores increased with time. The Paneth cells metaplasia and hyperplasia were observed on D14 until D28 (p<0.01 vs. control group) followed by an increased in the levels of IL-1b. The levels of a-defensins protein expression were initially increased (D7-D14; p<0.01 vs. control group) but then undergo reciprocal changes on D19 until D28 (p<0.01 vs. D7 and D14). Positive correlations between the degree of enterocolitis and Paneth cells number were detected in the sigmoid colon (r=0.42). CONCLUSION: Paneth cells underwent metaplasia and hyperplasia in the sigmoid colon of HSCR rats corresponding to an increase in the degree of enterocolitis, but not followed by an increase in the level of a-defensins as well as IL-1b, suggesting that there is an involvement of Paneth cells in the pathophysiology of enterocolitis due to HSCR.

410-5

Authors: Mardhatillah Sariyanti, Tiara Ayoe Andita, Noor Diah Erlinawati, Elvira Yunita, Ahmad Azmi Nasution, Kartika Sari, Nikki Aldi Massardi, Sylvia Rianissa Putri

BACKGROUND: Intestinal microbial dysbiosis and its metabolites can affect the immune activity of intestinal mucosal cells, causing insulitis and pancreatic β-cell death. Probiotic Lactobacillus acidophilus plays an important role in reducing inflammatory cytokines, hence improves oxidative stress that affects pancreatic β-cell apoptosis. Current study examined the feature of pancreatic histopathology affected by the administration of probiotic L. acidophilus in rats with type-1 diabetes mellitus (DM) induced by streptozotocin (STZ). METHODS: Twelve rats were induced by STZ at double dose of 50 mg/kgBB before administered with probiotic L. acidophilus at a dose of 1.5x10 8 or 1.5x10 9 CFU/mL/day, while other 4 rats were used as control. After 21 days of the L. acidophilus treatment, the average of fasting blood glucose (FBG) levels of rats were measured, then the pancreatic histopathology was assessed to evaluate the degree of insulitis in islet of Langerhans. RESULTS: The induction of STZ had been succeeded to increase blood glucose levels, which indicate DM condition. The highest FBG level after 21 days of treatment was found in DM group with glucose level of 512±81.51 mg/dL. The administration of probiotic L. acidophilus during 21 days treatment at both dose 1.5x10 8 and 1.5x10 9 CFU/mL/day significantly improved pancreatic histopathology (p=0.04 and p=0.034, respectively), with significant decrease on insulitis scores compared to DM group. CONCLUSION: The administration of L. acidophilus at both dose of 1.5x10 8 and 1.5x10 9 CFU/mL/day for 21 days can improve pancreatic histopathology of type-1 DM rats induced by STZ, therefore probiotic L. acidophilus may be potential as supplementation treatment for type-1 DM.

421-8

Authors: Wilya Isnaeni, Suryani As'ad, Mochammad Hatta, Saidah Syamsuddin, Fahrin Ramadan Andiwijaya, Hamsu Kadriyan

BACKGROUND: Cases of undernutrition, such as stunting and wasting, in Nusa Tenggara Barat (NTB), Indonesia, was found to be relatively high. Therefore, local government launched a golden generation program called GEN NTB to improve the quality of human resources by achieving a healthy, intelligent, devout, and productive generation in 2045. One of the genes known to be related with prosocial behavior is the oxytocin transferase (OXTR) gene. This study was conducted to determine the association between OXTR gene mRNA expression and prosocial behavior of the GEN NTB children. METHODS: This was an analytical observational case-control study involving 25 children as GEN NTB samples and 26 children as controls. Blood samples were tested for OXTR protein level with enzyme-linked immunosorbent assay (ELISA), and OXTR mRNA expression with real time polymerase chain reaction (RT-PCR). Prosocial behavior was characterized and determined by using a rating method, which valued from 1 to 4 for poor to very good behavior. RESULTS: The average OXTR protein levels of the GEN NTB group was 88.28 ng/mL, which were higher than the average OXTR protein levels of control group (2.41 ng/mL). According to fold change analysis, the OXTR mRNA expression in GEN NTB group was also higher than the control group (10.91 vs. 6.40). Interestingly, observations on the prosocial behavior of the GEN NTB group showed significantly higher rate values compared to the control group (17.3 vs. 8.0, p=0.034). Hence, these findings showed that the OXTR protein level and OXTR mRNA expression was correlated with the better prosocial behavior. CONCLUSION: Higher rating of prosocial behavior of the GEN NTB children is related to the higher OXTR mRNA expression levels. This might be attributed to the interventions of GEN NTB program that may elevate children's quality of life since early childhood.

436-41

Authors: Pieri Kumaladewi, Wirsma Arif Harahap, Bastian Nova, Irianiwati Widodo, Ramadhan Karsono, Ferry Sandra, Bethy Suryawathy Hernowo

BACKGROUND: Interaction between estrogen and estrogen receptor (ER) takes part in the regulation and differentiation of breast tumorigenesis. Some ERα polymorphisms, including ERα rs3798577, are reported to be associated with the risk and aggressiveness of breast carcinoma since the site was reported to be targeted by microRNA, which can further modulate the ERα expression. Hence, this study was conducted to disclose the possible role of ERα SNP rs3798577 on breast carcinoma patients. METHODS: Samples were taken from the post-mastectomy breast carcinoma tissues of female patients and screened based on the completeness of medical and histopathological records. DNA isolation was proceeded using real time-polymerase chain reaction (RT-PCR) then analyzed for high resolution melting (HRM). The nucleotide base sequence was then analyzed based on rs3798577 ERα polymorphism. ER immunohistochemistry test was carried out and counted quantitatively based on the staining intensity and the percentage of the stained cells. RESULTS: Out of 65 samples, there were 33 samples as wild type and 32 samples as variant type. Most variant and wild type had >80% ERα percentage. Most variant type had middle ERα intensity, while wild type had strong ERα intensity. Higher percentage of variant type (52.2%) was found with weak ERα histoscore, meanwhile higher percentage of wild type (52.4%) was found with strong ERα histoscore, but not significant (p=0.725). CONCLUSION: ERα rs3798577 variant type had a lower ERα intensity and weaker ERα histoscore compared to the wild type, suggesting that ERα rs3798577 polymorphism might play a role in breast carcinoma risk determination.