Human CALU(Calumenin) ELISA Kit

We attempted to investigate the relationship between hsa-let-7c and ANP32E, as well as their influence on the cells phenotype of lung adenocarcinoma. Expression of hsa-let-7c and prognostic values were assessed by bioinformatics analysis based on TCGA database. Quantitative real-time PCR and western blot was employed to measure relative expression of hsa-let-7c or ANP32E. The targeting relationship between let-7c and ANP32E was predicted by biological software and validated by dual luciferase reporter assay. With gene transfection technology, cell proliferation, invasion and migration were appraised by cell counting Kit-8, clone formation and Transwell assays. The results showed that hsa-let-7c was downregulated in lung adenocarcinoma. Downregulation of hsa-let-7c notably led to a poor survival. ANP32E was forecasted and confirmed as a directly target of hsa-let-7c, and was upregulated in lung adenocarcinoma. Furthermore, upregulation of ANP32E had a significant correlation with unsatisfactory survival. Meanwhile, the levels of ANP32E were negatively regulated by hsa-let-7c. Upregulation of hsa-let-7c remarkably suppressed the Calu-3 cell proliferation, invasion and migration, while ANP32E overexpression plasmids rescued the downtrend. Inversely, hsa-let-7c silencing in NCI-H209 cells presented the opposite outcomes. Collectively, hsa-let-7c shows an anti-tumor effect in lung adenocarcinoma by targeting ANP32E and is expected to be a potential therapeutic target for lung adenocarcinoma.

The major intracellular barriers associated with DNA delivery using nonviral vectors are inefficient endosomal/lysosomal escape and poor nuclear uptake. LAH4-L1, a pH responsive cationic amphipathic peptide, is an efficient DNA delivery vector that promotes the release of nucleic acid into cytoplasm through endosomal escape. Here we further enhance the DNA transfection efficiency of LAH4-L1 by incorporating nuclear localizing signal (NLS) to promote nuclear importation. Four NLSs were investigated: Simian virus 40 (SV40) large T-antigen derived NLS, nucleoplasmin targeting signal, M9 sequence, and the reverse SV40 derived NLS. All peptides tested were able to form positively charged nanosized complexes with DNA. Significant improvement in DNA transfection was observed in slow-dividing epithelial cancer cells (Calu-3), macrophages (RAW264.7), dendritic cells (JAWSII), and thymidine-induced growth-arrested cells, but not in rapidly dividing cells (A549). Among the four NLS-modified peptides, PK1 (modified with SV40 derived NLS) and PK2 (modified with reverse SV40 derived NLS) were the most consistent in improving DNA transfection; up to a 10-fold increase in gene expression was observed for PIG and PK2 over the unmodified LAH4-L1. Additionally PK1 and PK2 were shown to enhance cellular uptake as well as nuclear entry of DNA. Overall, we show that the incorporation of SV40 derived NLS, in particular, to LAH4-L1 is a promising strategy to improve DNA delivery efficiency in slow-dividing cells and dendritic cells, with development potential for in vivo applications and as a DNA vaccine carrier.