The diagnosis of endometrial cancer involves sequential, invasive tests to assess the thickness of the endometrium by a transvaginal ultrasound scan. In 6–33% of cases, endometrial biopsy results in inadequate tissue for a conclusive pathological diagnosis and 6% of postmenopausal women with non-diagnostic specimens are later discovered to have severe endometrial lesions. Thus, identifying diagnostic biomarkers could offer a non-invasive diagnosis for community or home-based triage of symptomatic or asymptomatic women. Herein, this study identified high-risk pathogenic nsSNPs in the NRAS gene. The nsSNPs of NRAS were retrieved from the NCBI database. PROVEAN, SIFT, PolyPhen-2, SNPs&GO, PhD-SNP and PANTHER were used to predict the pathogenicity of the nsSNPs. Eleven nsSNPs were identified as “damaging”, and further stability analysis using I-Mutant 2.0 and MutPred 2 indicated eight nsSNPs to cause decreased stability (DDG scores < −0.5). Post-translational modification and protein–protein interactions (PPI) analysis showed putative phosphorylation sites. The PPI network indicated a GFR-MAPK signalling pathway with higher node degrees that were further evaluated for drug targets. The P34L, G12C and Y64D showed significantly lower binding affinity towards GTP than wild-type. Furthermore, the Kaplan–Meier bioinformatics analyses indicated that the NRAS gene deregulation affected the overall survival rate of patients with endometrial cancer, leading to prognostic significance. Findings from this could be considered novel diagnostic and therapeutic markers.

This review begins with an introduction to the basic skeleton and classes of flavonoids. Studies on flavonoids have shown that the presence or absence of their functional moieties is associated with enhanced cytotoxicity toward cancer cells. Functional moieties include the C2-C3 double bond, C3 hydroxyl group, and 4-carbonyl group at ring C and the pattern of hydroxylation at ring B. Subsequently, the current knowledge on the chemistry, sources, pharmacology, and anticancer properties of diosmetin (DMT) and tamarixetin (TMT), two lesser-known methylated flavonoids with similar molecular structures, is updated. DMT is a methylated flavone with three hydroxyl groups, while TMT is a methylated flavonol with four hydroxyl groups. Both DMT and TMT display strong cytotoxic effects on cancer cell lines. Studies on the anticancer effects and molecular mechanisms of DMT included leukemia and breast, liver, prostate, lung, melanoma, colon, and renal cancer cells, while those of TMT have only been reported in leukemia and liver cancer cells. These findings suggest that flavones lacking the C3 hydroxyl group at ring C are more cytotoxic than flavonols having the C3 hydroxyl group. The in vitro and in vivo cytotoxic activities of DMT and TMT against cancer cells involve different molecular targets and signaling pathways. From this study, it is clear that little is known about the pharmacology and anticancer properties of DMT and TMT. The potentials for further research into these aspects of the two lesser-known methylated flavonoids are enormous.

One of the drivers for early carcinogenesis involves specific somatic point substitution mutations in the KRAS gene which damages its ability to conduct signal transduction. Although antibodies can be used for the targeting of KRAS antigen, their localization along the cell’s inner membrane serves as a barrier against the accessibility of the antibodies. This study describes the evaluation of two internalization strategies, namely the endocytosis-based cell penetrating peptide (CPP) approach and the adenoviral-based intrabody (IB) approach, for the delivery of an anti-mutant KRAS single-chain variable fragment (scFv) into the cell. Splicing by overhang extension polymerase chain reaction (SOE-PCR) was used for the fusion of scFv with an enhanced green fluorescence protein (eGFP) and Antennapedia-PTD (Antp), a cell penetrating signal peptide. The fused construct (Antp-scFv-eGFP) at a concentration of 0.085 mg/ml was expressed in E. coli (BL21), while recombinant adenoviral particles containing the scFv-eGFP gene were harvested from HEK 293 cells. Both SW480 and HeLa cells were treated with Antp-scFv-eGFP and recombinant adenoviral particles, and their eGFP localization and intensity were compared to determine their scFv binding efficiencies. The IB approach was shown to exhibit a 3-fold higher fluorescence signal intensity compared to the CPP approach. This proof-of-concept study demonstrated that both antigens for either screening, diagnostic approaches can be potentially adopted when targeting various intracellular or therapeutic purposes.