Introduction: Amyloid plaques, mainly comprising of amyloid-beta peptides derived from its precursor protein, are found deposited in hippocampal and entorhinal cortical regions of patients with Alzheimer’s disease (AD). However, none led to a complete understanding of the molecular mechanisms of the AD in order to generate a new therapy that would eradicate the disease effectively. Activation of pro-apoptotic pathway was found to be associated with the accumulation of amyloid precursor protein (APP). The objective of this study is to examine theeffects of APP overexpression on the Bcl-2 family proteins involving in pro-apoptotic pathway in neuronal cells. Methods: The experiment was first performed with the transfection of HEK 293T cells for generation of lentiviral vector system consisting APP plasmid followed by transduction of SH-SY5Y neuronal cells using lentivirus generated. Subsequently, western blot analysis was conducted to validate the APP overexpression in SH-SY5Y cells. Then, expression levels of Bcl-2 family proteins in the APP overexpressed cells were determined by western blot analysis. The statistical analysis was performed by Microsoft Excel with Student’s t-test. Results: APP overexpression in SH-SY5Y cells slightly upregulated the pro-apoptotic proteins including Bad, Bid, Bok and Puma but slightly downregulated Bcl-2, Bim and Bax. Conclusion: Our data suggest that APP overexpression regulated the Bcl-2-mediated pathway by a significant downregulation of Bim protein in neuronal cells.
Introduction: Human papillomavirus (HPV) is a small, non-enveloped double-stranded circular DNA virus. The high risk types of HPV are claimed to be responsible for over 99% of cervical cancers while the most common oncogenic genotypes 16 and 18, implicated in approximately 70% of all cervical cancers. HPV has six early genes (E1, E2, E4, E5, E6 and E7) that code for regulatory proteins involved in viral DNA replication, transcription control and cellular transformation. The E7 protein is responsible for the escape from cell cycle arrest in HPV infected cells by binding to the retinoblastoma protein (pRB) through its LXCXE binding motif. This inhibits the tumour suppressor protein from regulating the cell cycle progression from G1 to S phase. In this study, the pocket domain of pRB which is targeted by LXCXE is used as a target to design peptide inhibitors using in silico methods. Methods: Biovia Discovery Studio Visualizer and UCSF Chimera softwares were used in designing the peptides. Results: Two crystal structures: 1GUX and 4YOZ were superimposed and studied. The similar amino acid sequences which bind to LXCXE were cut to form the potential peptide inhibitors. Based on this, peptide 1 was selected for further in vitro analysis. The cytotoxicity of the peptide 1 was analysed on three cell lines: CaSki (HPV16+ cervical cancer cell), C33a (cervical cancer cell), and HaCaT (normal keratinocyte). As a result, the IC50 of one of the designed peptide inhibitor – peptide 1, on CaSki cell line was 180 μM. The inhibitory effect of the peptide 1 was also analysed validated using Western Blot using antibodies from the B-Myb and pRB family. The changes in cell cycle before and after treatment of the peptide inhibitor will be further monitored using a cell analyser. Conclusion: Peptide 1 had shown potential as an inhibitor of the HPV E7 protein based on the in silico analysis, but further functional studies are needed to validate its potential.
Introduction: Ruthenium compounds are widely studied for its biological activity. However, potent ruthenium drugs often have limited bioavailability due to its poor aqueous solubility. In order to assist the preparation of these hydrophobic compounds, a carrier or drug delivery agent is often introduced. Herein, we encapsulated a hydrophobic ruthenium polypyridyl complex [Ru(dppz)PIP]2+, (dppz = dipyrido-[3,2-a:20,30-c]phenazine, PIP = 2-phenylimidazo[4,5-f][1,10] phenanthroline) in mesoporous silica nanoparticles (MSN). Methods: The MSN was synthesized by using ionic liquid 1-hexadecylphenanthrolinium as a novel template and have 833.99 m2/g in surface area. The cytotoxicity of the synthesized ruthenium complex, MSN and MSN-loaded ruthenium was studied on Hela and A549 cancer cell lines. Results: MSN was non-toxic at lower dosage (
Introduction: Ruthenium polypyridyl complex (RPC), [Ru(dppz)2PiP]2+ or RuPiP, where dppz = dipyridophenazine, and PiP = 2-phenylimidazo[4,5-f][1,10]phenantroline has been shown to exhibit anticancer activities by stalling the replication fork progression in human cancer cell line, causing DNA double-strand break (DSB) leading to the initiation of DNA damage response (DDR). Poly (ADP-ribose) polymerase (PARP) enzymes are activated in response to DNA damage thus, RuPiP may be advantageously combined with the inhibitors of PARP to improve its efficacy in cancer cell killing. This study was conducted to investigate the cytotoxic effects of RuPiP and selected PARP inhibitor, NU1025, alone or in combination in vitro and the possible combinations in order to achieve synergism against three different cancer cell lines. Methods: Cell viability was determined by MTT assay based on established method and the combination index (CI) values were calculated using Chou and Talalay method. Results: Here, we reported that the treatment with RuPiP alone led to dose-dependent decreases in the cell viability meanwhile NU1025 exhibited no toxicity as a single agent. The CI values (
Introduction: Combination therapy to treat cancer have been demanded due to the complexity of the disease and to prevent resistance mechanisms commonly found in classic chemotherapeutic methods. Recently, we have reported that [Ru(dppz)2(PIP)]2+ (dppz = dipyridophenazine, PIP = 2-(phenyl)imidazo[4,5-f][1,10]phenanthroline) (RuPIP) immediately stalls replication fork progression in HeLa human cervical cancer cells. Co-incubation with a Chk1 inhibitor achieves synergistic apoptosis in cancer cells. These discoveries indicate that this class of compounds merit further investigation as anticancer drugs, especially within combinational therapy roles. However, information pertaining to the effects of combining ruthenium compounds with existing chemotherapeutic drugs remain scarce. This study aimed to investigate the possible synergistic cytotoxic effects of using RuPIP in combination with cisplatin on different cancer cell lines. Methods: A549, MCF7, Hela and T24 cells were treated with different concentrations of RuPIP or cisplatin alone, as well as different combinations of these two agents at a fixed ratio 1:1 over the course of 72 hr to assess their individual and combination effects. Cell viability was analysed using MTT assay. The combination index (CI) was calculated based on the Chou Talalay Method. Results: Single-agent treatment at 72 hr with RuPIP or cisplatin led to dose-dependent decreases in the viability of the A549, MCF7, Hela and T24 cells at 72 hr. Furthermore, increasing the concentrations of the combinations up to four folds of half maximal inhibitory concentration (IC50) statistically decrease the cell survival rates of A549 and MCF7 cells thus, displayed synergistic effects. Conclusion: Treatment of MCF7 and A549 cells with a combination of RuPIP and cisplatin showed a synergistic effect and thus are promising as a combination therapy for cancer treatment.
Introduction: Germacrone is a natural product isolated from Rhizoma curcuma with anti-tumor, anti-inflammatory, and anti-bacterial properties. Previous studies have demonstrated that Germacrone exhibits anti-tumor effect in breast and hepatoma cancer cell lines but the studies of its molecular mechanisms and anti-tumor properties in other cancers are not well studied. This study aims to investigate the anti-tumor effect of Germacrone on human skin, cervix, and gastric cancer cell lines and the molecular mechanism underlying the anti-tumor effect of Germacrone. Methods: A375 (skin malignant melanoma), AGS (gastric adenocarcinoma), and HeLa 229 (cervix adenocarcinoma) cell lines were employed for this research. Treatment of the cell lines with Germacrone has inhibited the cell proliferation in a dose-dependent manner as assessed by MTT assay. The cell lines were incubated with Germacrone for 24 hours followed by detection of the expression of BAX, BAK, p53, BCL2, MCL1, and BCL-XL using Real-time PCR. Results: Results from Real-time PCR has showed that pro-apoptotic gene BAK was highly expressed in all the human cell lines after the treatment with Germacrone. Furthermore, the expression of pro-apoptotic gene p53 were elevated in both A375 and HeLa 229 cell lines but not inAGS cell lines. The expression level of pro-survival genes BCL2 and MCL1 were found to be decreased in both AGS and A375 cell lines. Conclusion: In conclusion, Germacrone might be a potent anti-tumor drug candidate for Human Melanoma, Cervix Adenocarcinoma, and Gastric Adenocarcinoma by increasing the expression level of pro-apoptotic proteins BAK. Future studies will focus on studying the cytotoxicity effect of combination of Germacrone with standard chemotherapy drugs on Human Melanoma, Cervix Adenocarcinoma, and Gastric Adenocarcinoma.
Introduction: Constipation is affecting a quarter of human population at any one time in all age groups. However, a proper gamma scintigraphic study of whole GI transit is rarely performed in Malaysia due to the lack of suitable radiopharmaceutical. Hence, this study was taken to develop a suitable radiotracer formulation for gamma scintigraphy study of whole gastric-intestinal transit. Methods: The biocompatible polystyrene (PS) incorporated with 152Sm2O3 (5%, w/v) will be used to synthesize the radiotracer. The 152Sm-labelled PS particles was neutron activated to 153Sm in a nuclear reactor for 5 minutes. Characterization of the physicochemical properties, gamma spectrometry and in-vitro radiolabeling studies in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) were carried out to study the properties and stability of the radiotracer before and after neutron activation. Results: Scanning electron microscope (SEM) and particle size analysis showed that size, shape and surface morphology of the particles remained after neutron activation. The synthesized 153Sm-labelled PS radiotracer (100 mg) particles achieved an activity of 3.7 MBq after 46 hrs. As indicated by the gamma spectrometry result, there is no long half-life radioimpuirties present in the samples. The 153Sm-labelled PS particles achieved radiolabeling efficiency of more than 95% in both SGF and SIF over 72 hrs. Conclusions: A 153Sm-labelled radiotracer particles formulation has been successfully developed from biocompatible PS. The proposed formulation has the advantage of cheaper, easier to be produced and reduced radiation exposure to staff. Further studies are required to validate the in-vivo performance of 153Sm-labelled formulation for assessing GI motility and transit in clinical use.
Introduction: The Hedgehog (Hh) pathway serves as a major regulator in organ development, stem cell maintenance, cell proliferation, and cell differentiation. This pathway is highly regulated and aberrant activation will promote tumorigenesis. Hh pathway notably Sonic Hedgehog pathway was reported to be upregulated and promote tumorigenesis in various human malignancies including colorectal, gastric, lung, prostate, and breast. This review was aimed to discuss the current understanding of Hh pathway activation in different types of human cancers and discuss the development of the therapeutic applications targeting Hh pathway. Methods: A systematic review was conducted using the electronic research database PubMed Central (PMC) from 2014-2019. The search was limited to studies that are relevant to both Hh signalling pathway and human cancers. A total of 50 articles were selected and their references cited were searched and reviewed. Results: The results regarding the role of Hh signalling in pancreatic cancer and colorectal cancer are controversial with some reporting tumor promoting activities whereas others tumor suppressive activity. Besides, results from other studies suggesting that Hh signalling pathway plays an oncogenic role by inducing tumor cells proliferation, promoting metastasis and maintaining cancer stem cells in human cancers such as lung, stomach, and breast. To date, Glasdegib (PF-04449913) is the only Hh targeting small molecule inhibitor being studied at FDA Phase 3 clinical trial. Identification of the right tumors and minimization of the side effects remain as the main obstacles in the development of Hh signalling inhibitors. Conclusion: In conclusion, advancement in our understanding of Hh pathway has provided us opportunity to develop novel therapeutic strategies to fight human cancers with activated Hh pathway but more studies need to be conducted to solve the controversial regarding the role of Hh pathway in certain cancers.
Introduction: Hepatic radioembolization is a minimally invasive procedure involving intrarterial administration of radioembolic microspheres for the treatment of liver tumours. In this study, a biocompatible polystyrene (PS) microspheres formulation containing radioactive Samarium-153 (153Sm) was synthesized and tested. The 153Sm emits both diagnostic gamma energy and therapeutic beta radiation, renders the synthesized microspheres an ideal theranostic radioembolic agent for hepatic radioembolization. Methods: First, the 152Sm2O3 (20 – 50%, w/v) was encapsulated in PS microspheres using solid-in-oil-in-water solvent evaporation method. The 152Sm-labelled PS microspheres were then activated to 153Sm (Eβmax = 807.6 keV, half-life = 46.3 hours) via 152Sm (n,γ) 153Sm reaction in a nuclear reactor with a neutron flux of 2.0 x 1012 n.cm-2.s-1. Physicochemical characterization, gamma spectroscopy and in-vitro radiolabeling studies were carried out to study the properties and stability of the microspheres before and after neutron activation. Results: The 153Sm -labelled PS microspheres achieved a nominal activity of 4.0 GBq.g-1 after 6 hours of neutron activation. Scanning electron microscope (SEM) and particle size analysis show that the microspheres remained spherical with diameters within 15 – 60 μm after neutron activation. No long half-life radioimpurities were found in the samples as revealed by the gamma spectroscopy results. The 153Sm-labelled PS microspheres achieved radiolabeling efficiency of more than 95% in saline and blood plasma over 480 hours. Conclusion: A biocompatible 153Sm-radiolabelled PS microspheres formulation has been successfully developed. The formulation achieved desirable properties for theranostic treatment of liver tumours. The formulation is relatively cheaper, easier to be produced and more readily available.
Introduction: Cancer has become a major economic and societal burden. The National Cancer Registry of Malaysia (NCR) estimates that one in four Malaysian (1:4) will develop cancer by the age of 75. This project aims to develop a prototype named “Laser ablation needle” for tissue cauterization and percutaneous hyperthermia cancer therapy. Our ultimate goal is to develop a highly flexible, operator-friendly and cost-effective laser ablation needle for tissue cauterization and hyperthermia cancer therapy, hence to improve the overall cancer survival rate and quality of life among the cancer patient population. Methods: The laser ablation needle is a closed loop opto-electronic control system, consists of a 2 mm Fiber Bragg Grating (FBG) – optical fiber temperature sensor, a laser driven hot needle and a micro-controller. Based on real-time temperature input from the FBG sensor, the micro-controller can perform a dynamic PID control on laser intensity for a safe hyperthermia treatment. In the fabrication, a medical grade optical fiber with a diameter of 800 μm was used for laser delivery. The optical fiber was embedded inside a biocompatible resin-made needle and connected to a 450 nm high power blue laser diode. The FBG temperature sensor was incorporated in the needle for real-time temperature monitoring and control. Focal hyperthermia produced by the laser-driven hot needle was conducted on ex-vivo bovine liver. Results: The rise in temperature was recorded by increasing laser power. The temperature profile was obtained at each depth. Irreversible thermal denaturation during irradiation was captured. Conclusion: These preliminary results suggest that this technique can be applied safely and effectively for cancer treatment. The developed prototype comprised of the diode laser showed that it can deliver its energy via simple optical fiber. This laser is cheaper and much smaller than the conventional high power lasers used in other studies.
Introduction: To date, lung cancer has become increasingly prevalent and remains the leading cause of cancer-related death in both sexes, globally. Despite the advances of cancer treatment, systemic chemotherapy remains as the major treatment option for lung cancer. Nevertheless, the trend of chemotherapy resistance has restricted the efficacy of the chemotherapy treatment, thus leading the urge to develop an alternative chemotherapeutic agent which could give a promising treatment effect. Short peptides have acquired increasing interest as promising therapeutics due to its anticancer potential, rapid kinetics, high potency and low biocompatibility issue. In search of novel anti-cancer leads, the main objective of this study is to evaluate the in vitro antiproliferative properties of hybridized peptide analogues against human lung adenocarcinoma (A549) cell line. Methods: ND and DN analogues were designed based on two parent peptides, NDC1 and NDC2, through fragments hybridization approach. Modification of amino acid residues at specific positions of NDC1 and NDC2 was done at the C-terminal. Then, MTT assay was performed to examine the antiproliferative activities of NDC1, NDC2, NDs and DNs against A549 cells at concentrations ranging from 2-256μg/mL for 24 hours. Results: Findings obtained showed that the parental peptides, NDC1 and NDC2, exhibited IC50 values of 47.5±4.950μg/mL and 239±9.899μg/mL, respectively. All NDs showed excellent antiproliferative activities with IC50 values ranging from 22-71μg/mL. Nevertheless, all DNs did not display antiproliferative activity when tested up to 256μg/mL. We speculated that increased valine and isoleucine with decreased aspartic acid composition in NDs might be associated with their intermediate cytotoxicity strength, comparing with the parent peptides. However, the location of other amino acids in the peptide sequence should still be further investigated as it contributes to the peptide structure, hence leading to its selectivity and potency. Conclusion: As a conclusion, NDs could be further explored to develop a potent anti-cancer therapeutic drug.
Introduction: Colon cancer is one of the leading cause of cancer death and current treatments often bring about undesired toxicities and resistance. Hence targeted therapeutic regimens for cancer are developed. Anticancer agent incorporated with copper has been synthesized to selectively target cancer cells that are reported to take up more copper compared to normal cells. Cu(SBCM)2 synthesized from the condensation of s-benzyldithiocarbazate and 3-aetylcoumarin was demonstrated to exhibit anti-proliferative effect towards MCF-7 cells and MDA-MB-231 breast cancer cells via cell cycle arrest and apoptosis. However, the mode of cell death of Cu(SBCM)2 on colon cancer cells has not been explored. This study investigated the anti-cancer properties of Cu(SBCM)2 on HT-29, colorectal cancer cell line. Methods: The growth inhibition of the copper complex was determined using MTT assay and xCELLigence real time cell monitoring analysis. Results: Cu(SBCM)2 was shown to inhibit the growth of HT-29 cells significantly in time- and dose- dependent manner with IC50 of 25.23 ± 8.22 uM at 48 hours. Morphological studies using inverted light microscope indicating Cu(SBCM)2-treated HT-29 cells displayed characteristics of apoptosis such as cellular shrinkage and membrane blebbing. Cell cycle analysis was carried out using flowcytometer and Cu(SBCM)2 was found to induce G2M cell cycle arrest at 24 and 48 hours. ROS assay was carried out to determine the involvement of oxidative stress on Cu(SBCM)2 treated HT-29 cells. Nevertheless, results indicated Cu(SBCM)2 significantly suppressed the formation of ROS compared to control. Conclusion: In summary, Cu(SBCM)2 shows promising potential in cancer therapy against colon cancer cells.
Introduction: This paper is aimed to evaluate the feasibility of computed tomography (CT) based thermometry for radiofrequency ablation (RFA) through the investigation on the effects of principal CT acquisition parameters to the CT number. Methods: The effects of CT acquisition parameters (tube voltage, tube current, gantry rotation time, and CT reconstructed slice thickness), as well as metal artefacts on CT number were evaluated by using liver tissue equivalent polyacrylamide (PAA) phantom. The correlation between CT number and tissue temperature from 37 0C to 80 0C was studied with the use of PAA phantom with optimum CT acquisition parameters. Results: The CT numbers show insignificant changes under variation of tube voltages, tube currents, gantry rotation time, and CT reconstructed slice thickness respectively. The CT number difference obtained for each series of the variations are less than 2 HU, which indicates the changes in the CT acquisition parameters has insignificant effects on the CT number shift. On the other hand, the CT reconstructed image is able to improve the metal artefact caused by RF electrode through the increases of CT reconstruction condition. In this paper. A linear regression analysis on the CT number and temperature suggested the CT number is inverse linearly proportional to temperature, with a CT thermal sensitivity of –0.521 ± 0.061 HU/0C (R2 = 0.998). Conclusion: In summary, the results show that assessment of CT thermometry is feasible with the use of current CT technology as it produces high reproducibility and stable CT measurement, which is proved to be independent for CT acquisition parameters.
Introduction: In an on-going research, the endophyte Aspergillus sp. HAB10R12 have been selected for detailed chemical investigation after its crude ethyl acetate extract showed promising anticancer properties with high selectivity. Methods: The former was determined on two cancer cell lines namely, HCT116 and MCF-7 (IC50 = 0.05 and 0.04 μg/mL, respectively) and one non-tumor cell line HeLa (IC50 = 10.5 μg/mL). Results: The result indicates the secondary metabolites produced by the fungus are 200 folds more selective towards cancer cells over normal cells, calling for an immediate detailed investigation of their composition. Preliminary chemical analysis of the crude extract using LC-MS, NMR and UHPLC-UV showed the presence of an uncommon group of diterpene pyrones (NF00659 A1, B1 and A3), previously isolated once with only partial characterization reported. Consequently, large scale isolation of secondary metabolites was carried out and led to the identification of four of the previously isolated diterpene pyrones. Conclusion: The isolation, characterization, relative stereochemistry analysis, and a plausible biosynthesis of the diterpene pyrone compounds is presented herein.
Introduction: Short peptides have acquired increasing interest as promising therapeutics, particularly as anti-cancer alternatives during the recent years. They have been reported to demonstrate incredible anti-cancer potentials through targeting signalling transduction pathways, as well as modulation of cell cycle, tumour suppressor proteins and transcription factors. Peptides are primarily of interest due to its rapid kinetics, high potency and low biocompatibility issue. In search of novel anticancer leads, the main objective of this study is to evaluate the in-vitro antiproliferative properties of hybridized peptides against human hepatocellular carcinoma (HepG2) cells. Methods: Two series of hybridized peptides, ND and DN analogues were designed based on two parent peptides, NDC1 and NDC2, through fragments hybridization approach. Modification of amino acid residues at specific positions of NDC1 and NDC2 was done at the C-terminal. Then, MTT assay was performed to examine the antiproliferative ranging from 2-256µg/mL for 24 hours. Results: The parent peptide, NDC1 showed an IC50 value of 87±3.786 µg/mL at 24 hours while NDC2 did not display antiproliferative activity against HepG2 cells. ND1-4 showed higher toxicity against Vero cells compared to HepG2 cells while ND5 did not exhibit antiproliferative activities against both cell lines. In contrary, DN1 and DN4 was found to exhibit antiproliferative activity against HepG2 cells, with IC50 values 170±60.883µg/mL and 170±60.883µg/mL, respectively. Meanwhile, both these hybridized peptides showed minimal toxicity against Vero with IC50 values >256µg/mL. In contrast, DN2, DN3 and DN5 showed minimal antiproliferative activity against HepG2 with IC50 values >256µg/mL. Conclusion: Among the ND and DN hybridized peptides, two hybridized peptides, DN1 and DN5, showed potential anti-proliferative activities against HepG2 with minimal toxicity against Vero. Nevertheless, their activity has been diminished as compared to NDC1 and hence, can be further improved.
Introduction: Colorectal cancer is the second most common cause of death in women, and the third most common cause in men. The main treatments available following surgery are chemotherapy and radiotherapy which are not always effective and have side effects. Modern medicine aims to develop targeted cancer therapies that are efficient and cause less side effects to patients. However, this approach requires a thorough understanding of the molecular events that cause cancer cell to grow and divide in order to identify suitable targets. The process of translating the findings into clinical studies can be high cost and technically demanding. However, development of a tissue microarray (TMA), allows immunohistochemical (IHC) staining of multiple cases simultaneously, thereby greatly reducing costs and time. Methods: A TMA was produced from approximately 400 cases of colorectal cancer, along with collection of associated clinical and pathological data. Sections from the TMA were tested for quality by staining with haematoxylin and eosin (H & E), in addition to IHC markers to molecularly classify the colon cancers. Results: The cores from the 384 cases of cancer were successfully transferred to 18 recipient TMA blocks. H & E staining showed good morphological preservation of the cases, reflecting the tumour in the donor blocks. IHC testing was able to successfully classify cases into distinct molecular groupings. Conclusions: The development of a TMA of colorectal cancers provides a valuable tool for the efficient and subsequent molecular classification of colorectal cancer using immunohistochemistry.
Introduction: Copper complexes can be developed as targeted agent for cancer due to increased uptake of copper by cancer cells for angiogenesis. Our previous published data showed that copper complex Cu(SBCM)2 induced apoptosis towards triple-negative MDA-MB-231 breast cancer cells. However, its effect towards other breast cancer subtype remains unknown. Current study was performed to explore the cytotoxicity of Cu(SBCM)2 towards oestrogen- receptor positive MCF-7 breast cancer cells. Methods: MTT assay was employed to study the growth inhibition of Cu(SBCM)2 towards MCF-7 breast cancer cells and human non-cancerous MCF10A breast cells. Morphological changes of Cu(SBCM)2-treated MCF-7 cells was observed under inverted light microscope. Induction of cell cycle arrest and apoptosis were accessed by flow cytometry. The expression of wild-type p53 protein was evaluated by western blotanalysis. Intracellular reactive oxygen species of MCF-7 treated with Cu(SBCM)2 was detected using DCFHDA assay. The cells were then co-treated with Cu(SBCM)2 and antioxidants to evaluate the involvement of ROS in the cytotoxicity of Cu(SBCM)2. Molecular docking study was performed to determine the interaction of Cu(SBCM)2 with DNA, DNA topoisomerase I, and human ribonucleotide reductase. Results: Cu(SBCM)2 induced G2/M phase cell cycle arrest and apoptosis in MCF-7 cells possibly via upregulation of p53 wild-type protein. Cu(SBCM)2 was less toxic towards MCF10A cells. Increased level of intracellular ROS was not detected in MCF-7 cells after treatment with Cu(SBCM)2. However, N-acetylcysteine antioxidant enhance the cytotoxicity of Cu(SBCM)2 in MCF-7 cells. Cu(SBCM)2 showed the greatest affinity for DNA topoisomerase I in comparison to DNA and human ribonucleotide reductase. Conclusions: Cu(SBCM)2 has a potential to be developed as a targeted agent for breast cancer.
Introduction: In Malaysia, about 800 children are diagnosed with cancer, a globally a dreadful disease, each year, and osteosarcoma accounts for approximately 3% of such cancer. The early detection of the type and extent of bone cancer is important for effective management through surgery. But, the presence of soft tissue edema around a neoplasm can interfere with accurate local tumor staging and subsequent surgical planning. However very scanty research is done on this; none of the past studies focused specifically on the incidence and quantity of extraosseous edema and its impact. Our interdisciplinary retrospective study with objectives to study the presence of soft tissue edema adjacent to the tumors in the extremities, characterize their pattern and distribution involved 82 patients of wide range of age attending the Hospital Universiti Sains Malaysia with a histologically proven tumor or tumor-like lesion of bone. Methods: This paper emphasizes avoiding misinterpretation of such edema and subsequent over-staging of musculoskeletal tumor. We exclusively used 1T-Magnetic Resonance Imaging due to its excellent resolution. All cases were imaged (T1W, T2W, T1W fat suppressed Gadolinium enhanced, and STIR images) by using 1.5 Tesla MRI unit. STIR images permit study of larger volume of abnormal tissue than spin echo images. Results: Peritumoral edema in 5 cases, Paratumoral edema in 11 patients and mixed type in 45 cases were found including 10 benign tumors. Overall, 5 malignant lesions did not show any sift tissue edema! All the data were analyzed and interpretation and charecterisation of the edema was done by an experienced radiologist. The findings were correlated with histopathology examination results. Conclusion: In conclusion, accurate definition of the local extent of a bone tumor and exploration of soft tissue edema is required to maximize the success of diagnostic work-up and staging prior to biopsy and subsequent interventions while minimizing the amount of tissue removed.
Introduction: The mortality rate of glioma patients particularly with glioblastoma multiforme (GBM) remains high even with advancements in the multimodality treatment. This is partly due to chemoresistance of the glioma cells towards drug treatment which finally reduced the survival of GBM patients. In this study, we determined the chemosensitisation and oncogenic characteristics of ZFP36L2 in LN18 GBM cells using RNA interference (RNAi). Methods: Meta-analysis of microarray datasets was used to identify the druggable genes responsive to GBM chemosensitivity. Subsequently, the genes were validated using RNAi screening [pooled small interference RNA (siRNA)]. Temozolomide- resistant LN18 cells were used to evaluate the effects of gene silencing on chemosensitisation to the sub-lethal dose (1/10 of IC50) of temozolomide. Assays such as cell viability, proliferation, migration and invasion and apoptosis assays were carried out. The apoptosis pathway underlying chemosensitisation by ZFP36L2 siRNA was determined using a human apoptosis array kit. Statistical analyses were performed using one-way analysis of variance. Results: ZFP36L2 was identified as a potential marker of GBM based on the meta-analysis and RNAi screening. ZFP36L2 knockdown lead to 1) Apoptosis induction (p < 0.05) 2) Reduced cell migration (p < 0.05) 3) Reduced up to 82% of cell invasion (p < 0.01) and 4) Decreased cellular proliferation in siZFP36L2-treated LN18 cells. Downstream analysis showed that the sub-lethal dose of temozolomide caused major upregulation of BCL2-associated X, apoptosis regulator (BAX). Conclusion: ZFP36L2 is oncogenic and chemosensitive thus may contribute to gliomagenesis through cell proliferation, migration, invasion and apoptosis. RNAi therapy in combination with chemotherapy treatment such as temozolomide may serve as potential therapeutic approach in the future.
Introduction: Gynura crepioides (G. crepioides) belongs to the Asteraceae family and native to Southeast Asia, especially Indonesia. Gynura genuses are well known for their antioxidant properties. Hence, the current study aimed to study the effect of temperature on polyphenololic content and antioxidant properties on G. crepioides leaves extract. Methods: The extraction of G. crepioides leaves was carried out by Ultrasound-assisted extraction (UAE) method for 60 minutes by using ethanol (70 %) at three different temperatures 25°C and 50°C and 75°C. The total polyphenolic content (TPC) was investigated by using Folin-Ciocalteu assay, whereas the antioxidant potential (AOP) was determined by using phosphomolybdenum assay, and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Results: The results from the study showed that TPC of G. crepioides extracted at 25°C was significantly (p < 0.05) lower than 50°C and 75°C with 8.45 ± 0.31 mg GAE/g, 35.83 ± 1.22 mg GAE/g, 35.90 ± 0.78 mg GAE/g respectively. However, the results from AOP has demonstrated lower value of 70.62 ± 0.74 mg AAE/g at 75°C compare to 77.67 ± 0.26 mg AAE/g at 25°C and 81.53 ± 0.68 mg AAE/g at 50°C. From DPPH assay results revealed that extraction temperature at 50°C has EC50 (p < 0.05) lowest value of 92.64 ± 0.56 µg/mL compared to G. crepioides extracted at 25°C (98.50 ± 1.18 µg/mL) and 75°C (101.72 ± 9.09 µg/mL). An excellent correlation exhibited between TAC and DPPH radical scavenging assays with r = 0.95 and r2 = 0.89. Present study reveals that UAE method with 70% ethanol, 60 minutes extraction time at 50°C temperature is an optimum to extract highest phenolic content and antioxidants from G. crepioides leaves. Conclusion: It was concluded from the study that extraction temperature would affect the extraction of phytochemicals from plants in turn it affects the biological activities.