BackgroundBreast and ovarian cancers are the most prevalent cancers and one of the leading causes of death in Indian women. The healthcare burden of breast and ovarian cancers and the rise in mortality rate are worrying and stress the need for early detection and treatment.MethodsWe performed amplicon sequencing of 144 cases who had breast/ovarian cancer disease (total 137 cases are patients and seven are tested for BRCA1/2 carrier) Using our custom designed gene panel consisting of 14 genes, that are associated with high to moderate risk of breast and ovarian cancers. Variants were called using Torrent Variant Caller and were annotated using ThermoFisher’s Ion Reporter software. Classification of variants and their clinical significance were identified by searching the variants against ClinVar database.ResultsFrom a total of 144 cases, we were able to detect 42 pathogenic mutations in [40/144] cases. Majority of pathogenic mutations (30/41) were detected in BRCA1 gene, while (7/41) pathogenic mutations were detected in BRCA2 gene, whereas, (2/41) pathogenic mutations were detected in TP53 gene and BRIP1, PALB2, and ATM genes respectively. So, BRCA genes contributed 88.09% of pathogenic mutations, whereas non-BRCA genes contributed 11.91% of pathogenic mutations. We were also able to detect 25 VUS which were predicted to be damaging by in silico prediction tools.ConclusionEarly detection of cancers in the Indian population can be done by genetic screening using customized multi-gene panels. Indications of our findings show that in the Indian population, apart from the common BRCA genes, there are other genes that are also responsible for the disease. High frequency mutations detected in the study and variants of uncertain significance predicted to be damaging by in silico pathogenicity prediction tools can be potential biomarkers of hereditary breast and ovarian cancer in Indian HBOC patients.
Nowadays CHK2 mutation is studied frequently in hereditary breast and ovarian cancer patients in addition to BRCA1/BRCA2. CHK2 is a tumor suppressor gene that encodes a serine/threonine kinase, also involved in pathways such as DNA repair, cell cycle regulation and apoptosis in response to DNA damage. CHK2 is a well-studied moderate penetrance gene that correlates with third high risk susceptibility gene with an increased risk for breast cancer. Hence before planning large population study, it is better to scrutinize putative functional SNPs of CHK2 using different computational tools. In this study, we have used various computational approaches to identify nsSNPs which are deleterious to the structure and/or function of CHK2 protein that might be causing this disease. Computational analysis was performed by different in silico tools including SIFT, Align GVGD, SNAP-2, PROVEAN, Poly-Phen-2, PANTHER, PhD-SNP, MUpro, iPTREE-STAB, Consurf, InterPro, NCBI Conserved Domain Search tool, ModPred, SPARKS-X, RAMPAGE, Verify-3D, FT Site, COACH and PyMol. Out of 78 nsSNP of human CHK2 gene, seven nsSNPs were predicted functionally most significant SNPs. Among these seven nsSNP, p.Arg160Gly, p.Gly210Arg and p.Ser415Phe are highly conserved residues with conservation score of 9 and three nsSNP were predicted to be involved in post translational modification. The p.Arg160Gly and p.Gly210Arg may interfere in phosphopeptide binding site on FHA conserved domain. The p.Ser415Phe may interfere in formation of activation loop of protein-kinase domain and might interfere in interactions of CHK2 with ligand. The study concludes that mutation of serine to phenylalanine at position 415 is a major mutation in native CHK2 protein which might contribute to its malfunction, ultimately causing disease. This is the first comprehensive study, where CHK2 gene variants are analyzed using in silico tools hence it will be of great help while considering large scale studies and also in developing precision medicines related to these polymorphisms in the era of personalized medicine.
In current study, the phytochemical, antioxidant and cytotoxic effects of methanol extracts of Vitex negundo, Lantana camara, Bauhinia variegata and Bauhinia racemosa were reported. Methanol extract of medicinal plants (V. negundo, L. camara, B. variegata and B. racemosa) were extracted using different extraction methods and then tested for their antioxidant activity by 2,2-diphenylpicrylhydrazyl, superoxide scavenging method, chelation of metal ions and cytotoxic potency using MTT assay on human cancer cell lines. In different extraction methods, highest yield was obtained with hot extraction method. Total phenolic and flavonoid contents were found higher in methanol extract of V. negundo having 173.3±1.54 mg gallic acid equivalent/g DW and 24.41±1.5 mg quercetin equivalent/g DW contents, respectively. Leaves of V. negundo have maximum IC 50 value of 73±2.76 µg/ml and 350±1.23 µg/ml in 2,2-diphenylpicrylhydrazyl and iron chelating activity compared to all other methanol extracts. B. racemosa leaves have higher superoxide scavenging effect with IC 50 value of 223±1.23 µg/ml. V. negundo and L. camara leaves showed pronounced cytotoxic effect against HELA and KB human cancer cell lines with LD 50 value of 222±3.35 and 188.69±1.4 µg/ml, respectively. The present study shows that V. negundo and L. camara may be a probable source of natural antioxidant and as an anticancer drugs.
Objective: The study was planned to investigate antioxidant and anticancer activities with the preliminary phytochemical analysis of methanolic extracts of Vitex negundo (V. negundo), Lantana camara (L. camara) and Bauhania variegata (B. variegata) plants leaf extracts. Methods:Phytochemical evaluation was performed for all the extracts, as per the standard methods. In vitro antioxidant activities were performed by using DPPH (2,2-Diphenyl-1-Picrylhydrazyl), ABTS (2, 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid) and FRAP (Ferric reducing antioxidant power assay) method and compared with standard antioxidants. The anticancer activity of plant extract was assessed using MTT colorimetric assay. Results:The study of preliminary phytochemical proved the existence of alkaloids, flavonoids and phenolic types of phytochemicals in high amount. Methanolic extract of L. camara shows minimum IC50 value for DPPH assay (48.75±2.34 µg/ml) and FRAP assay (274.66±3.65 µg/ml). In ABTS assay B. variegata extract exhibit minimum IC50 value (60.48±3.01 µg/ml). Lower the IC50 value of extract, higher the effectiveness of the plant. Methanolic extract of all plants methanolic extracts showed anticancer activity against SH-SY-5Y cells (human neuroblastoma cell) but V. negundo was more effective against SH-SY-5Y cells with IC50 value (209 µg/ml) compared to remaining extracts. Conclusion:The current finding accomplished the in vitro activities, so that plant could be a superior source of antioxidant and anticancer drugs. But further in vivo assessment was needed before adding it into the pharma industry.
Cancer is one of the most dreadful diseases globally and it appears to be due to extreme free radical damage, which eventually causes damage to the DNA, lipids and protein. In cell cycle, growth and division of normal cells occur in an well-ordered manner, however in cancerous cells, defective caspase-mediated cell death (apoptosis) leads to increased cell proliferation [1] . Some proteins manage the temporal order within the cell cycle highly regulated to confirm that division of cells occur only when required. Inaccuracy in this directive becomes the characteristic of cancer [2,3] . Tumour cells evolved mechanisms to resist cell death and this has motivated researchers to investigate many plants, which might selectively induce apoptosis in cancer cells [3,4] . It was reported that presence of some phytochemicals in herbs could be the main reason for antitumor activity exerted through inducing apoptosis in cancer cells [5][6][7][8] . In programmed cell death, activation of caspases (cysteinyl, aspartate-specific proteases) leads to activation of target substrates within the
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