Genetic diversity analysis in the Hypericum perforatum populations in the Kashmir valley by using inter-simple sequence repeats (ISSR) markers

Assessment of genetic variability among the Hypericum perforatum populations is critical to the development of effective conservation  strategies in the Kashmir valley. To obtain accurate estimates of genetic diversity among and within populations of H. perforatum, inter-simple sequence repeats (ISSR) markers were used. The study was aimed to check, whether ISSR fingerprinting may be a useful tool for studying genetic variations among H. perforatum populations in the Kashmir valley (India). A total of 15 ISSR primers were tested with the 20 genotypes of H. perforatum. The ten informative primers were selected and used to evaluate the degree of polymorphism and genetic relationships within and among all the H. perforatum populations. ISSR of 20 genotypes analysis yielded 98 fragments that could be scored, of which 71 were polymorphic, with an average of 7.1 polymorphic fragments per primer. Number of amplified fragments varied in size from 150 to 1650 bp. Percentage of polymorphism ranged from 60% to a maximum of 100%. Resolving power ranged from a minimum of 7.7 to a maximum of 14.3. Shannon indexes ranges from 0.166 to 0.389 with an average of 0.198 and Nei’s genetic diversity (h) ranges from 6.98 to 9.8. Estimated value of gene flow (Nm = 0.579) indicated that there was limited gene flow among the populations. The genetic diversity (Ht) within the population of 0.245 was clearly higher than that of among population genetic diversity (Hs= 0.115), indicating an out-crossing predominance in the studied populations. Analysis of molecular variance by ISSR markers indicated that over half of the total variation in the studied populations (58%) could be accounted for by differences among the 8 divisions, with a further 42% being accounted for by the variation among populations within a division.The dendrogram grouping the populations by unweighted pair-group method with arithmeticaverages (UPGMA) method revealed eight main clusters. In conclusion, combined analysis of ISSR markers and hypericin content is an optimal approach for further progress and breeding programs.Keywords: Hypericum perforatum (St. John's Wort), inter-simple sequence repeats (ISSR) markers, unweighted pair-group method with arithmetic averages (UPGMA), Nei’s genetic diversityAfrican Journal of Biotechnology, Vol. 13(1), pp. 18-31, 1 January, 201

Utility of Serum miR-125b as a Diagnostic and Prognostic Indicator and Its Alliance with a Panel of Tumor Suppressor Genes in Epithelial Ovarian Cancer.

MicroRNAs (miRNAs) have been found to be dysregulated in epithelial ovarian cancer (EOC) and may function as either tumor suppressor genes (TSGs) or as oncogenes. Hypermethylation of miRNA silences the tumour suppressive function of a miRNA or hypermethylation of a TSG regulating that miRNA (or vice versa) leads to its loss of function. The present study aims to evaluate the impact of aberrant microRNA-125b (miR-125b) expression on various clinicopathological features in epithelial ovarian cancer and its association with anomalous methylation of several TSGs. We enrolled 70 newly diagnosed cases of epithelial ovarian cancer, recorded their clinical history and 70 healthy female volunteers. Serum miR-125b levels were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and the methylation status of various TSGs was investigated by methylation specific PCR. ROC curves were constructed to estimate the diagnostic and prognostic usefulness of miR-125b. The Kaplan-Meier method was applied to compare survival curves. Expression of miR-125b was found to be significantly upregulated (p<0.0001) in comparison with healthy controls. The expression level of miR-125b was found to be significantly associated with FIGO stage, lymph node and distant metastasis. ROC curve for diagnostic potential yielded significant AUC with an equitable sensitivity and specificity. ROC curves for prognosis yielded significant AUCs for histological grade, distal metastasis, lymph node status and survival. The expression of miR-125b also correlated significantly with the hypermethylation of TSGs. Our results indicate that DNA hypermethylation may be involved in the inactivation of miR-125b and miR-125b may function as a potential independent biomarker for clinical outcome in EOC

Association of miR-125b expression with the promoter hypermethylation of a panel of tumour suppressor genes.

<p>(A)DAPK1 (B) p16 (C) RASSF1A (D) PTEN (E) BRCA1 (F) p14. Positive and negative depicts the presence or absence of promoter hypermethylation of the specified tumour suppressor gene respectively.</p

Association of serum miR-125b and clinicopathological characteristics in epithelial ovarian cancer patients with respect to A)Stage, (B)Lymph node metastasis and (C)Distant metastasis status.

<p>Association of serum miR-125b and clinicopathological characteristics in epithelial ovarian cancer patients with respect to A)Stage, (B)Lymph node metastasis and (C)Distant metastasis status.</p

AUC for ROC curve corresponding to the diagnostic value of miR-125b in EOC.

<p>AUC for ROC curve corresponding to the diagnostic value of miR-125b in EOC.</p

Clinical implications of cytosine deletion of exon 5 of P53 gene in non small cell lung cancer patients

Aim: Lung cancer is considered to be the most common cancer in the world. In humans, about 50% or more cancers have a mutated tumor suppressor p53 gene thereby resulting in accumulation of p53 protein and losing its function to activate the target genes that regulate the cell cycle and apoptosis. Extensive research conducted in murine cancer models with activated p53, loss of p53, or p53 missense mutations have facilitated researchers to understand the role of this key protein. Our study was aimed to evaluate the frequency of cytosine deletion in nonsmall cell lung cancer (NSCLC) patients. Methods: One hundred NSCLC patients were genotyped for P53 (exon5, codon168) cytosine deletion leading to loss of its function and activate the target genes by allele-specific polymerase chain reaction. The P53 cytosine deletion was correlated with all the clinicopathological parameters of the patients. Results and Analysis: 59% cases were carrying P53 cytosine deletion. Similarly, the significantly higher incidence of cytosine deletion was reported in current smokers (75%) in comparison to exsmoker and nonsmoker. Significantly higher frequency of cytosine deletion was reported in adenocarcinoma (68.08%) than squamous cell carcinoma (52.83%). Also, a significant difference was reported between p53 cytosine deletion and metastasis (64.28%). Further, the majority of the cases assessed for response carrying P53 cytosine deletion were found to show faster disease progression. Conclusion: The data suggests that there is a significant association of the P53 exon 5 deletion of cytosine in codon 168 with metastasis and staging of the disease

Sequence of mature miR-125b primer used.

<p>Sequence of mature miR-125b primer used.</p

Area under curve (AUC) of receiver operating characteristic (ROC) for miR-125b corresponding to (A)Tumor grade (B)Metastasis (C)Lymph node status and (D)Survival of EOC patients.

<p>Area under curve (AUC) of receiver operating characteristic (ROC) for miR-125b corresponding to (A)Tumor grade (B)Metastasis (C)Lymph node status and (D)Survival of EOC patients.</p

Kaplan Meier survival curve with respect to (A) fold change (B) mucinous and serous histopathological subtypes.

<p>Kaplan Meier survival curve with respect to (A) fold change (B) mucinous and serous histopathological subtypes.</p

Serum expression of miR-125b.

<p>(A) Dot plot showing the relative expression of miR-125b in patients and controls (B) ROC curve for miR-125b exhibiting its diagnostic potential in epithelial ovarian cancer.</p