Characterization of Eighty-Eight Single-Nucleotide Polymorphism Markers in the Manila Clam Ruditapes philippinarum Based on High-Resolution Melting (HRM) Analysis.

Single-nucleotide polymorphisms (SNPs) are the most commonly used DNA markers in population genetic studies. We used the Illumina HiSeq4000 platform to develop single-nucleotide polymorphism (SNP) markers for Manila clam Ruditapes philippinarum using restriction site-associated DNA sequencing (RAD-seq) genotyping. Eighty-eight SNP markers were successfully developed by using high-resolution melting (HRM) analysis, with a success rate of 44%. SNP markers were analyzed for genetic diversity in two clam populations. The observed heterozygosity per locus ranged from 0 to 0.9515, while the expected heterozygosity per locus ranged from 0.0629 to 0.4997. The value of FIS was estimated to be from -0.9643 to 1.0000. The global Fst value was 0.1248 (p < 0.001). After Bonferroni correction, 15 loci deviated significantly from the Hardy-Weinberg equilibrium (p < 0.0006). These SNP markers provide a valuable resource for population and conservation genetics studies in this commercially important species.

Tetrandrine triggers apoptosis and cell cycle arrest in human renal cell carcinoma cells.

Tetrandrine is a cytotoxic compound capable of exerting remarkable antitumor activity against many cancer cells in vitro and in vivo. However, little is known about its effect on human renal cell carcinoma (RCC). In the present study, using RCC 786-O, 769-P and ACHN cell lines as the model system, we demonstrated the anticancer effect of tetrandrine against RCC and clarified its underlying mechanisms. Tetrandrine treatment showed growth inhibitory effects on RCC cells in a time- and dose-dependent manner. Additionally, flow cytometric studies revealed that tetrandrine was capable of inducing G1 cell cycle arrest and apoptosis in RCC cells. Mechanically, activation of caspase-8, caspase-9, and caspase-3 and increasing expression of cell cycle regulatory protein p21(WAF1/CIP1) and p27(KIP1) were observed in tetrandrine-treated RCC cells. This study provides the first evidence that tetrandrine triggered apoptosis and cell cycle arrest in RCC 786-O, 769-P and ACHN cells in vitro; these events are associated with caspase cascade activation and upregulation of p21 and p27. Our results thus provide rational evidence supporting the application of tetrandrine as a novel therapeutic agent against RCC in the clinical setting.