RESUMEN
Natural plant products have been widely used in controlling cancer with fewer or no side effects and the use of plant extracts as complementary to synthetic medicine is gaining increased popularity. Members of Andrographis plants possess important medicinal properties. In the present study, anti-cancerous properties of Andrographis nallamalayana (AN) were tested on A375 and B16F10 skin melanoma cancer cell lines. The leaf extracts of AN significantly reduced the cell viability and cell survival of skin cancer cell lines, achieved by MTT assay and clonogenic assays, respectively. Further, TUNEL assays revealed that AN extracts induces the apoptosis. Western blot analysis revealed that AN leaf extracts reduced the expression of Bcl-2, an anti-apoptotic protein and induced the expression of proapoptotic molecules such as Bcl-2 associated death promoter protein (BAD), Bcl-2 associated X protein (BAX) and cleaved caspase-3. Moreover, the qRT-PCR and western blot analysis demonstrated the reduced expression of G2/M phase proteins cdk1, cyclin B1 and increased expression of p53, cyclin-dependent kinase 1 inhibitor, p21. Further, immunofluorescence analysis revealed that AN reduced the NF-κB nuclear translocation, luciferase reporter assays demonstrated reporter gene activation. qRT-PCR assays showed that AN significantly reduced the expression of NF-κB target genes. The results concluded that the extracts of AN exhibited significant anti-proliferative, anti-inflammatory and pro-apoptotic activities on melanoma skin cancer cell lines.
RESUMEN
Familial Hypertrophic Cardiomyopathy (FHC) is an autosomal dominant disorder affecting the cardiac muscle and exhibits varied clinical symptoms because of genetic heterogeneity. Several disease causing genes have been identified and most code for sarcomere proteins. In the current study, we have carried out clinical and molecular analysis of FHC patients from India. FHC was detected using echocardiography and by analysis of clinical symptoms and family history. Disease causing mutations in the ß-cardiac myosin heavy chain (MYH7) and Myosin binding protein C3 (MYBPC3) genes were identified using Polymerase Chain Reaction-Deoxyribose Nucleic Acid (PCR-DNA) sequencing. Of the 55 patient samples screened, mutations were detected in only nineteen in the two genes; MYBPC3 mutations were identified in 12 patients while MYH7 mutations were identified in five, two patients exhibited double heterozygosity. All four MYH7 mutations were missense mutations, whereas only 3/9 MYPBC3 mutations were missense mutations. Four novel mutations in MYBPC3 viz. c.456delC, c.2128G>A (p.E710K), c.3641G>A (p.W1214X), and c.3656T>C (p.L1219P) and one in MYH7 viz. c.965C>T (p.S322F) were identified. A majority of missense mutations affected conserved amino acid residues and were predicted to alter the structure of the corresponding mutant proteins. The study has revealed a greater frequency of occurrence of MYBPC3 mutations when compared to MYH7 mutations.