ARMS-PCR Based SNP Analysis of MTHFR C677T Allele Using Syber Green in Pancreatic Tumour.

In human, carcinoma of pancreas, a rare disease and mortality rate is quite high in Indian population. Epidemiological studies support the hypothesis that folate metabolism regulate DNA stability and prevent cancer. Because folate have been linked to dietary supplement and defect in folate metabolism have been increase risk of developing cancer shows adverse effect on health conditions. In the present study we have assess methylene tetrahydrofolate reductase gene polymorphism (MTHFR) 677 C → T using ARMS PCR based SNP analysis using Syber green in the cases of pancreatic tumour to determine the “risk factor”. Interestingly, our findings reveals that 33.0% frequency (one case) showing mutation of MTHFR 677 TT genotype (rare type) in homozygous condition with Tm value 82.50°C for mutant 677T allele shifted to 677C allele (83.50°C).Two cases (66%) showing CC (wild type) allele and Tm value 82.83°C for 677C allele. In MTHFR 677TT is a rare mutation and individuals show very low enzymatic activity due to the substitution of alanine to valine. The study further continue to confirm the mutation by visualization on agarose gel electrophoresis of the same PCR product, again showing (Lane- 2) the band of 50 kb of mutant 677TT allele in the same case, suggesting an relevant role of folate metabolism and subsequent impairment of aberrant DNA methylation during carcinogenesis with increasing “high risk” for the development of carcinoma due to allele TT mutation of MTHFR. However, large samples size is required to further confirm the association.

Cell cycle inhibitory activity of Piper longum against A549 cell line and its protective effect against metal-induced toxicity in rats.

Anticancer potential of Piper longum fruit against human cancer cell lines (DU-145 prostate, A549 lung, THP-1 leukemia, IGR-OVI-1 ovary and MCF-7 breast) as well as its in vitro and in vivo biochemical efficacy in AlCl3-induced hepatotoxicity were evaluated in the rats. Dried samples were extracted with several solvents using soxhlet apparatus. Flavonoid content in chloroform, benzene, ethyl alcohol and aqueous extracts of fruit was 19, 14, 12 and 11 μg quercetin equivalent/mg of sample, respectively. Hexane extracts exhibited 90-92% cytotoxicity against most of the test cell lines (A549, THP-1, IGR-OVI-1 and MCF-7), while benzene extract displayed 84-87% cytotoxicity against MCF-7, IGR-OV-1 and THP-1 cell lines. Among extracts, hexane, benzene and acetone extracts demonstrated considerable cytotoxicity (91-95%) against A549 (lung cancer) cell line in Sulforhodamine B dye (SRB) assay. Cell cycle analysis revealed that hexane, benzene and acetone extracts produced 41, 63 and 43% sub-G1 DNA fraction, demonstrating cell cycle inhibitory potential of these extracts against A549 cell line. Chloroform, ethyl alcohol and aqueous extracts displayed 71, 64 and 65% membrane protective activity, respectively in lipid peroxidation inhibition assay. P. longum fruit extracts also ameliorated AlCl3-induced hepatotoxicity, as indicated by alterations observed in serum enzymes ALP, SGOT and SGPT activity, as well as creatinine and bilirubin contents. In conclusion, study established the cytotoxic and hepatoprotective activity in P. longum extracts.

Synthesis, biological evaluation and molecular docking of aryl hydrazines and hydrazides for anticancer activity.

Aryl hydrazine and hydrazide analogues were synthesized based on p-tolyl hydrazine, isolated as a breakdown product of a secondary metabolite from the mushroom, Agaricus bisporus, and tested to be highly active molecule than 5-fluorouracil in in vitro anticancer studies. The synthesized analogues were tested for anticancer activity using NCI protocol. Anolgues 12 and 15 emerged as molecules with significant in vitro anticancer activity. Molecular docking study revealed the binding orientations of aryl hydrazines and hydrazides analogues in the active sites of thymidylate synthase.