ABSTRACT
Objective: To research the mechanism of ebracteolatain A against breast cancer cells, screening the genes with significant changes using second-generation sequencing, and explore the anti-breast cancer mechanism of action of ebracteolatain A at the transcriptomics level. Methods: The acetyl phloroglucinol compound ebracteolatain A was extracted from Euphorbia ebracteolata, interferencing with MCF7 cells (luminal A type of breast cancer cells) to observe differential gene expression between the interfered cells and normal cells. High-throughput transcriptome sequencing and data analysis were performed on three groups of control groups and three experimental groups using Illumina Hi-Seq sequencing technology. Results: A total of 123 656 848, 123 974 262 available reads were obtained in the control group and experimental group, respectively, the reads on the reference genome were 119 762 214, 119 881 622, respectively, accounting for 96.85% and 96.69% of the total; Two groups of transcriptome controls were available: the total number of differential genes was 1 695, of which 770 were up-regulated, 925 were down-regulated, and 3 874 genes were clearly annotated. Bio-enrichment analysis was carried out by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). GO analysis found that these 3 874 genes mainly involved in biological processes (1 270), cell composition (1 322) and molecular function (1 282), 45 subcategories of three major categories, including cell growth and development, signaling protein activity, membrane and regulation of gene expression. KEGG analysis revealed that the differentially expressed genes involved 263 signaling pathways; The main metabolic pathways were: PI3K-Akt signaling pathway, MAPK signaling pathway, carbohydrate metabolism, myocardial system and cellular reproductive system and etc. Conclusion: The results showed that 1 695 differential genes were screened and identified by Illumina Hi-Seq sequencing technology, and the relationship between the genes of ebracteolatain A and MCF7 cells was further understood, which provided some theoretical cornerstones for breast cancer treatment.
ABSTRACT
The purpose was to investigate the effect of phytohemagglutinin (PHA) on proliferation and cytotoxicity of cytokine-induced killer (CIK). Peripheral blood mononuclear cells (PBMNCs) from healthy donors were divided into two groups. Cells were resuspended and maintained in complete medium containing of 10% autologous plasma. CIK cells were cultured by traditional method in group one. The other group cells were added PHA to stimulate PBMNCs for 24 hours, then cultured like incubating CIK cells. Their cytotoxicity to different target cells was evaluated by (51)Cr release assay. The results showed that the proliferation multiples of CIK and PHA-CIK cells were both high, however, the latter was much higher than CIK with significance (P < 0.05). Cells in each group cells showed high cytotoxicity. At the same high effector/target ratio PHA-CIK cells cytotoxicity was stronger than CIK cells when targets were K562 cells or acute leukemia cells (P < 0.05). In conclusion, PHA-CIK cells exhibit stronger proliferation and cytotoxicity than CIK cells, and the result provides an experimental basis for biotherapy.