RESUMO
Objective To understand the influence of different conditions to the potential phenotypes of cells, through the transcriptome sequencing and analysis for two cancer cell lines with identical genetic background from different sources, so as to provide an experimental basis for rigorous cell model-based drug screening. Methods Human cervical cancer (HeLa) cell and human liver cancer (HepG2) cell lines, newly introduced from ATCC (HeLa-ATCC and HepG2-ATCC) and a previously laboratory-stored (HeLa-PLS and HepG2-PLS) cell lines were used in the present study. The second generation high-throughput sequencing platform Illumina Novoseq was used for transcriptome sequencing, and the Gene Ontology (GO) function and pathway enrichment analysis based on the GO data base and the Kyoto Encyclopedia of Genes and Genomes (KEGG) data base, fusion gene analysis and analysis of mutation sites were performed for the differentially expressed genes (DEG). Meanwhile, the sequencing and analytical work was also performed for evaluating the effect of different culture times on the transcriptome of the ATCC-sourced HeLa cells. Results Total 7366 and 8786 DEGs in the HeLa and HepG2 cell lines were found between the HeLa-ATCC and HeLa-PLS cells and between the HepG2-ATCC and HepG2-PLS cells, respectively, HeLa and HepG2 cells, which were classified into 519 and 778 metabolic pathways by the GO function and pathway enrichment analysis, respectively. The DEGs in the HeLa cells were mainly enriched in the extracellular matrix tissues, metabolic pathways, cancer signaling pathways, cytokines and their interaction systems with receptors. The DEGs in the HepG2 cells were mainly enriched in the extracellular matrix tissues, neural active ligands and their interaction systems with receptors, cancer signaling pathways and mitogen-activated protein kinase (MAPK) signaling pathways. The cancer signaling pathway was attenuated in the HeLa-PLS cell line, while the cancer signaling pathway was enhanced in the HepG2-PLS cell line, accompanied with changes in the neuroactive factor-receptor binding activity and cytokine-receptor binding activity. On the other hand, significant changes on the genes related to the ion channels, the cell membrane receptors, such as G protein-coupled receptors, and the intracellular calcium signaling pathways found in the HeLa-ATCC cells within 6-24 hours after the cells were fully adhered. Conclusion Compared with the HeLa-ATCC and HepG2-ATCC cells, the transcriptome of the HeLa-PLS and HepG2-PLS cells showed significant changes, involving a wide range of cellular functions. Cell transcriptome sequencing might likely provide useful informations for understanding cell identity and controlling experimental conditions for drug screening and evaluation.
RESUMO
Objective To discover antitumor drugs showing a synergistic effect with the cannabinoid receptor agonist sildenafil mesylate (WIN55212-2), so as to provide a new strategy for potential drug combinations for improving the life quality of cancer patients. Methods Firstly, the antitumor activity was tested for the combination of cannabinoid receptor 1 (CB1R) receptor agonist WIN55212-2 with each of 25 antitumor drugs using three tumor cell lines with high CB1R, HepG2, DU145 and HCT-8, by highthroughput assay. Then, the in vitro tumor colony-forming assay and 3D tumor spheroid assay were conducted to confirm the synergistic effect for the effective drug combination. Flow cytometry was used to investigate the effect of the synergistic drug combination on the apoptosis and cell cycle progression. Results Three drugs showed a synergistic inhibitory effect on the proliferation of tested tumor cells by combining with WIN55212-2, and among them, the combination of exemestane with WIN55212-2 displayed best effect, which showed a dose-dependent synergistic antitumor effect in the in vitro tumor colony-forming test and 3D tumor spheroid assay (CI＜1).Compared with the single-exemestane treatment, the combination of exemestane with WIN55212-2 significantly increased the apoptosis of HepG2 cells (P＜0.01) and caused G2/M phase arrest of the HepG2 cells. Conclusion The study is the first to report that the combination of exemestane with WIN55212-2 showed a synergistic anti-tumor activity on HepG2 cells, which was likely related to the promotion of apoptosis and induction of cell cycle arrest.