Identification of therapeutic targets and mechanisms of tumorigenesis in non-small cell lung cancer using multiple-microarray analysis.

Lung cancer is the most commonly occurring cancer attributed to the leading cause of cancer-related deaths globally. Non-small cell lung cancer (NSCLC) comprises 85% to 90% of lung cancers. The survival rate of patients with advanced stage NSCLC is in months. Moreover, the underlying molecular mechanisms still remain to be understood.We used 2 sets of microarray data in combination with various bioinformatic approaches to identify the differentially expressed genes (DEGs) in NSCLC patients.We identified a total of 419 DEGs using the Limma package. Gene set enrichment analysis demonstrated that "Citrate cycle (TCA cycle)," "RNA degradation," and "Pyrimidine metabolism" pathways were significantly enriched in the NSCLC samples. Gene Ontology annotations of the 419 DEGs primarily comprised "glycosaminoglycan binding," "cargo receptor activity," and "organic acid binding." Kyoto Encyclopedia of Genes and Genomes analysis revealed that DEGs were enriched in pathways related to "Malaria," "Cell cycle," and "IL-17 signaling pathway." Protein protein interaction network analysis showed that the hub genes constituted of CDK1, CDC20, BUB1, BUB1B, TOP2A, CCNA2, KIF20A, CCNB1, KIF2C, and NUSAP1.Taken together, the identified hub genes and pathways will help understand NSCLC tumorigenesis and develop prognostic markers and therapeutic targets against NSCLC.

Tubeimoside-1 inhibits the proliferation and metastasis by promoting miR-126-5p expression in non-small cell lung cancer cells.

Tubeimoside-1 (TBMS1) possesses broad anticancer activities, including the cytostatic and anti-angiogenesis effects in lung cancer. However, the effect of TBMS1 on the metastasis of non-small cell lung cancer (NSCLC) cells and the potential underlying mechanism remain unclear. In the present study, a cell counting kit-8 assay revealed that TBMS1 suppressed the proliferation of NCI-H1299 cells significantly, particularly following 48 h of treatment. Further studies showed that TBMS1 notably enhanced the apoptosis, and inhibited the migration and invasion of NCI-H1299 cells upon treatment for 48 h. A total of 14 NSCLC tissues and 14 normal adjacent tissues were collected, reverse transcription-quantitative polymerase chain reaction revealed decreased expression of microRNA (miR)-126-5p in NSCLC tissues compared with adjacent NSCLC tissues, which was reversed following TBMS1 administration in NCI-H1299 cells. The overexpression of miR-126-5p induced by TBMS1 was demonstrated to target and downregulate vascular endothelial growth factor (VEGF)-A. Simultaneously, the expression of VEGF-R2 was reduced notably, along with a significant declined in the phosphorylation levels of dual specificity mitogen-activated protein kinase kinase 1 and extracellular signal-regulated kinase (ERK)1/2. Overall, the aforementioned results indicated that TBMS1 inhibited the proliferation and metastasis, and promoted the apoptosis of NCI-H1299 cells, which may be mediated by overexpressing miR-126-5p, which inactivates the VEGF-A/VEGFR2/ERK signaling pathway. Therefore, TBMS1 may be a promising drug for prevention and treatment of NSCLC.

Antitumor effects of Tubeimoside-1 in NCI-H1299 cells are mediated by microRNA-126-5p-induced inactivation of VEGF-A/VEGFR-2/ERK signaling pathway.

Tubeimoside-1 (TBMS1), a triterpenoid saponin isolated from the tuber of Bolbostemma paniculatum (Maxim) Franquet, serves an universal suppressive role in multiple cancer types, including lung cancer. However, the mechanism involved in non­small cell lung cancer (NSCLC) cells by which TBMS1 elicits its antitumor effects is not yet comple-tely understood. The present study indicated that 10 µmol/l TBMS1 significantly enhanced apoptosis and notably blocked the migration and invasion of NCI­H1299 cells. These effects were reversed following transfection with miR­126­5p inhi-bitor into TBMS1­treated NCI­H1299 cells. Vascular endo-thelial growth factor-A (VEGF­A) is a target gene for miR­126­5p. Notably, results suggested that the downregulated VEGF­A and VEGFR­2 in TBMS1­treated NCI­H1299 cells were upregulated after inhibiting miR­126­5p, and overexpression of VEGF­A or VEGFR­2 could significantly reduce apoptosis and promote the migration and invasion of TBMS1­treated NCI­H1299 cells. Furthermore, TBMS1 combined with TBHQ (an ERK activator) dramatically suppressed TBMS1­induced apoptosis and stimulated TBMS1­reduced migration and invasion in NCI­H1299 cells, suggesting that TBMS1 inhibits the ERK signaling pathway and represses the growth and metastasis of NCI­H1299 cells. Further study demonstrated that either inhibiting miR­126­5p or overexpressing VEGF­A and VEGFR­2 in TBMS1­treated NCI­H1299 cells elevated the mRNA expression levels and phosphorylation levels of MEK1, as well as ERK. To conclude, TBMS1 increases miR­126­5p expression, whereas overexpressing miR­126­5p inactivates VEGF­A/VEGFR­2/ERK signaling pathway, which ultimately actuates the pro­apoptotic and anti­metastatic effects in NCI­H1299 cells. Therefore, the present findings provide a theoretical foundation for TBMS1 as a potential candidate in NSCLC treatment.