The improvement of in vivo model (Balb/c mice) for cervical carcinogenesis using diethylstilbestrol (DES).

Cervical cancer is the most common gynecological cancer and one of the major causes of female cancer-related death worldwide particularly in developing countries. Thus far, there are a few in vivo models have been developed in investigating this type of cancer. In this study, we induced cervical cancer in Balb/c mice by exploiting the carcinogenic property of diestylstilbestrol (DES). The Balb/c pregnant mice were given subcutaneous (SC) injection of 67µg/kg body weight of DES on GD 13, and the mice gave birth approximately at gestation day 19-22. Female offspring were reared and the body weight was recorded once weekly. The female offspring were sacrificed at age of 5 months. Upon termination, blood was collected in a plain tube via cardiac puncture and the reproductive tracts were collected and weighed. The reproductive tract sections were stained using H&E for observation of pathological changes. The progression of disease state was monitored by measuring the level of serum interleukin (IL-6) using the Mouse IL-6 ELISA Assay Kit (BD OptEIA™, USA). All parameters were compared with Not-induced group. The outcome of this study demonstrated a significant difference in body weight gain, reproductive organ weight, diameter of cervix and the level of serum IL-6 in the Induced group as compared to the Not-induced group (P<0.05). Histopathological findings revealed the presence of adenosis only in the Induced group. It shows that DES could be employed as an agent to induce cervical carcinogenesis in animal model. In addition to that, new potential anti-cancer agents from various sources could be further evaluated using this technique.

Dillenia suffruticosa exhibited antioxidant and cytotoxic activity through induction of apoptosis and G2/M cell cycle arrest.

ETHNOPHARMACOLOGICAL RELEVANCE: Dillenia suffruticosa (Family: Dilleniaceae) locally known as Simpoh air has been reported to be used traditionally to treat cancerous growth. Therefore, the present study was attempted to investigate the antioxidant and cytotoxic properties of different parts (root, flower, fruit and leaf) of D. suffruticosa extracts. METHODS AND MATERIALS: In this study, direct solvent extraction (aqueous and methanol) from different parts of D. suffruticosa (root, flower, fruit and leaf) were carried out. Antioxidant activities of D. suffruticosa extract were determined by using DPPH, ABTS FRAP and ß-carotene bleaching assays. Cytotoxicity and cell cycle arrest of the active extract were determined using MTT assay and flow cytometer, respectively. Sequential solvent extraction (hexane, DCM, EtOAc, and MeOH) were also carried out in root of D. suffruticosa to further evaluate the antioxidant and cytotoxic activity of the different solvent extracts. RESULTS: Methanol (MeOH) root extract showed the highest TPC, antioxidant and cytotoxic activities (especially towards HeLa) compared to others (P<0.05). Based on the results, sequential solvent extraction (hexane, DCM, EtOAc and MeOH) was carried out in the roots of D. suffruticosa. MeOH extract exhibited the highest antioxidant activities among others and significantly correlated (P<0.05) with TPC, suggesting the important contribution of phenolic compounds to its antioxidant activity. On the other hand, the DCM and EtOAc exhibited higher cytotoxic activity to selected cancer cells (HeLa, MCF-7, MDA-MB-231, A549 and HT29) compared to others. In short, there is no established correlation between antioxidant and cytotoxic activities of D. suffruticosa extracts indicating that an agent with high antioxidant activities will not necessarily possesses good cytotoxic activities in return. Qualitative phytochemical screening of D. suffruticosa extracts suggested the presence of saponins, triterpenes, sterols, and polyphenolic compounds which are believed to contribute to the cytotoxic activities. CONCLUSION: It is suggested that the cytotoxicity of the active extracts in HeLa was due to the induction of apoptosis and cell cycle arrest at G2/M.

Antibacterial and biofilm inhibition activities of Melastoma malabathricum stem bark extract against Streptococcus mutants

Aim: This study was to determine the antibacterial activity of Melastoma malabathricum stem bark acetone extract (MMSBAE) against Streptococcus mutans. Methodology and results: Antibacterial activity of the extract was determined by minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), biofilm formation, adherence inhibition, time kill studies and effect on S. mutans membrane integrity. MIC and MBC values of MMSBAE were 1.25 and 5 mg/mL, respectively. Time kill studies showed that reduction of colony forming unit in treated cells is 3 log10 after 10 h of treatment (p ˂ 0.05). The extracts reduced 50% biofilm and adherence activity of S. mutans at 1.88 mg/mL. The effect on S. mutans membrane integrity after exposure to MMSBAE for 90 and 120 min was determined by measuring leakage of cell content at 2 different wavelengths of 260 nm and 280 nm. In leakage assay, the percentage of absorbance (260 nm) in treated cell material showed 57% at 90 min and 60% at 120 min which is higher than negative control (<20%) but less than positive control (100%). The percentage absorbance of treated cell material (280 nm) was 61% at 90 min and 63% at 120 min. Identification of compound in MMSBAE was done by gas chromatography mass spectrometry (GCMS). Ten compounds were identified in the MMSBAE with some of them important in antimicrobial activity such as ethyl ester, undecene, and gamma sitosterol. Conclusion, significance and impact of study: MMSBAE showed excellent bactericidal and antibacterial activities against S. mutans. The antibacterial mode of action of MMSBAE is suggested to be the disruption of the S. mutans membrane structure. The MMSBAE significantly inhibited biofilm and adherence activities of S. mutans in dose dependent manner (p ˂ 0.05). MMSBAE has potential in the development of antibacterial agent with anti-biofilm and anti-adherence activities.