The Indonesian Biomedical Journal

358-65

Authors: Habibah Faroque, Nur Soleha Aria, Nor Amira Muzilah, Hamid Ali Nagi Al-Jamal, Mohd Fadliyazid Ab. Rahim, Yasuhiro Nakamura, Hironobu Sasano, Abdullah Azmahani

BACKGROUND: The checkpoint kinase 1 (CHEK1) and glutamine-fructose-6-phosphate aminotransferase 1 (GFPT1) genes have been reported to have a crucial role in carcinogenesis in colorectal cancer (CRC). However, their association with the pathogenesis of CRC remains unclear. This study was conducted to study the expression of CHEK1 and GFPT1 genes in adenoma and adenocarcinoma CRC patients’ whole blood samples compared to the healthy controls. METHODS: A comparative cross‑sectional study to examine the expression of CHEK1 and GFPT1 genes were conducted in 6 colorectal adenoma and 6 colorectal adenocarcinoma patients along with 6 healthy controls. Blood samples were taken from subjects, and CHEK1 and GFPT1 genes were analyzed by using reverse transcription quantitative polymerase chain reaction (RT-qPCR). RESULTS: CHEK1 gene expression in blood samples has the highest in adenoma patients meanwhile for the GFPT1 gene expression has the highest in adenocarcinoma patients. Higher CHEK1 gene expression were also found in adenoma patients compared to healthy controls (p=0.040) and adenocarcinoma patients (p=0.025). Besides, at the 5% level of significance, the median GFPT1 gene expression was higher in colorectal adenocarcinoma patients compared to colorectal adenoma patients and healthy controls. CONCLUSION: CHEK1 and GFPT1 may function as potential regulators in adenoma and adenocarcinoma and measuring their expression might be a potential tool to determine the CRC progression and could be further explored as blood diagnostic biomarkers for CRC patients.

375-82

Authors: Christiana Tri Nuryana, Tiara Puspita Agustin, Sofia Mubarika Haryana, Yohanes Widodo Wirohadidjojo, Nur Arfian

BACKGROUND: Ultraviolet B (UVB) induces skin photoaging by reducing collagen deposition via impairment of the TGF-β/Smad signaling pathway. Achatina fulica mucus (AFM) is a native medicine acting as vehicle of anti-aging ingredients. The present investigation examined the effect of AFM on UVB-induced fibroblast photoaging by assessing TGF-β, Smad3, and Smad7 mRNA expressions. METHODS: AFM was extracted from A. fulica using electrical shock and freeze-dried into a powder. Normal human dermal fibroblast (NHDF) cultures were irradiated with/without 100 mJ/cm2 UVB and treated with/without 10% platelet-rich plasma or different concentrations of AFM: 3.9 μg/mL in AF3 group; 15.625 μg/mL in AF15 group, and 62.5 μg/mL in AF62 group. The mRNA expressions of TGF-β, Smad3, and Smad7 in NHDF were evaluated by quantitative polymerase chain reaction. RESULTS: TGF-β mRNA expressions in the AF3 (0.85±0.01), AF15 (0.94±0.02) and AF62 (1.64±0.03) groups were significantly higher (p<0.05) compared with that in the UVB group (0.55±0.04). Moreover, Smad3 expressions in the AF3 (1.42±0.25), AF15 (1.89±0.13), and AF62 (2.50±0.31) groups were significantly higher (p<0.05) compared with that in the UVB group (0.57±0.08). Furthermore, Smad7 expressions in the AF3 (1.57±0.18), AF15 (0.87±0.03), and AF62 (0.25±0.09) groups were significantly lower (p<0.05) than that in the UVB group (2.57±0.06). CONCLUSION: AFM ameliorates UVB-induced fibroblast photoaging by upregulating the TGF-β/Smad3 expressions and downregulating Smad7 expression.

391-9

Authors: Bogi Pratomo Wibowo, Handono Kalim, Husnul Khotimah, Hidayat Sujuti, Ettie Rukmigarsari, Nabila Erina Erwan

BACKGROUND: Colorectal cancer (CRC) is reported as the third most frequently diagnosed cancer worldwide. Salmonella infection plays a role in developing this cancer, which is chronically related to the avirulence protein A (AvrA) effector protein produced by the bacteria. This study was conducted to point out the effect of AvrA Salmonella on the occurrence of CRC through the regulation of TLR4/NF-κB/β-catenin/TGF-β expressions in the azoxymethane (AOM)/dextran sodium sulfate (DSS) CRC model. METHODS: A randomized control group post-test-only study was conducted using male Balb/c mice with 30-gram body weight (10-12 weeks), which were divided into three groups, namely the negative control (normal mice without any treatment), positive control (AOM/DSS-treated mice), treatment 1 (T1) (AOM/DSS-treated mice + AvrA Salmonella) groups. Colon tissue was collected and then prepared for immunohistochemistry staining using TLR4, β-catenin, NF-κB, TGF-β, and Ki67 antibodies, whereas apoptotic cells were stained using TUNEL assay. RESULTS: The expressions of TLR4, β-catenin, NF-κB, TGF-β, Ki67, and apoptosis percentage indicated significant differences among the three groups, which statistically showed p<0.05 in all observed parameters. The mean of all parameters was far more significant in the T1 group than in the negative and positive control groups. CONCLUSION: The findings revealed that AvrA Salmonella could increase the expressions of TLR4, β-catenin, NF-κB, TGF-β, and Ki67 and decrease the apoptotic percentage. Thus, AvrA Salmonella influences CRC tumorigenesis through TLR4/NF-κB/β-catenin/TGF-β and is suggested as a potential target in future preventive and curative management for CRC.

411-9

Authors: Michael Kevin, Nyoman Suci Widyastiti, Selamat Budijitno, Yan Wisnu Prajoko, Neni Susilaningsih

BACKGROUND: Andrographis paniculata has anti-inflammatory, anti-cancer, and immunomodulatory effects. Intercellular adhesion molecule-1 (ICAM-1) and E-selectin are released in inflammatory conditions due to sepsis. In sepsis-associated fatalities, E-selectin is strongly expressed on pulmonary microvasculature and ICAM-1 is strongly expressed on pulmonary microvasculature endothelial cells, pulmonary macrophages, and lymphocytes. Therefore, this study was conducted to analyze the protective effect of A. paniculata leaf extract against lung injury in lipopolysaccharide (LPS)-induced sepsis mouse model. METHODS: Thirty male Wistar rats were divided into 1 control group and 4 sepsis groups. The control group was fed ad libitum for 14 days, followed by an intraperitoneal injection of 5 mg/kgBW 0.9% NaCl on day-21. While the sepsis groups were fed ad libitum and orally administered 0, 200, 400, and 500 mg/kgBW of A. paniculata leaf extract, respectively, for 14 days. All four sepsis groups were intraperitoneally injected with 5 mg/kgBW LPS on day-21. Mice termination and measurement of ICAM-1 and E-selectin were performed on day-25. The immunohistochemical examination of ICAM-1 expression was performed using ICAM-1/CD54 antibody, meanwhile E-selectin expression was examined using primary antibody E-Selectin (D-7) rabbit IgG antibody. RESULTS: All groups given A. paniculata leaf extract showed lower expression of ICAM-1 (p<0.001) and E-selectin (p<0.001) than the group without extract. A. paniculata leaf extract 500 mg/kgBW gives the best effectiveness compared to other doses. CONCLUSION: A. paniculata leaf extract has a protective effect against lung injury by lowering the expression of ICAM-1 and E-selectin in lung tissue.

429-36

Authors: Pande Kadek Aditya Prayudi, I Gde Sastra Winata, I Nyoman Gede Budiana, Kade Yudi Saspriyana, I Nyoman Bayu Mahendra, Ketut Suwiyoga

BACKGROUND: The need for clinically useful biomarkers which can predict the surgical outcome after primary debulking surgery (PDS) in patients with advance ovarian cancer (AOC) is really important. Insulin-like growth factor-binding protein 2 (IGFBP2) is the main binding protein expressed by ovarian cancer cells, which plays a prominent role in promoting proliferation, driving invasion, and suppressing apoptosis. This study was conducted to assess the performance of IGFBP2 in predicting the surgical outcome after PDS in patients with AOC. METHODS: Twenty-four subjects with AOC (Stage IIIc/IV) who underwent PDS were recruited consecutively. Clinicopathologic data were obtained from subjects' medical records. Blood samples were withdrawn form each subject and preoperative level of IGFBP2 were measured using enzyme-linked immunosorbent assay (ELISA). Multivariate analysis was employed to test the performance of multiple predictors of surgical outcome. RESULTS: Eighteen patients (75%) had suboptimal outcome after PDS. Mean IGFBP2 level was significantly higher in the suboptimal group (1157.5±359.9 ng/mL vs. 679.1±504.5 ng/mL, p=0.018). In bivariate model, higher preoperative level of IGFBP2 predict the suboptimal outcome with good accuracy (AUC: 0.796, sensitivity: 83.3%, specificity: 83.3%, p=0.033, optimal threshold level 870 ng/mL). Higher IGFBP2 level was associated with higher risk of suboptimal outcome, although IGFBP2 was not an independent risk factor (adjusted OR: 5.0, 95% CI: 0.43-57.9, p=0.198). CONCLUSION: IGFBP2 is a novel and promising biomarker for surgical outcome prediction following PDS in AOC patients. Since it is predictive for suboptimal outcome, patients with higher preoperative level of IGFBP2 needs more thorough preoperative evaluation as well as meticulous surgical technique to optimize the surgical outcome.