cDNA microarray analysis of the effect of cantharidin on DNA damage, cell cycle and apoptosis-associated gene expression in NCI-H460 human lung cancer cells in vitro

Abstract

Cantharidin (CTD) induces cytotoxic effects in different types of human cancer cell; however, to date, there have been no studies on the effects of CTD on gene expression in human lung cancer cells and the potential associated signaling pathways. Therefore, the present study aimed to investigate how CTD affects the expression of key genes and functional pathways of human H460 lung cancer cells using complementary DNA microarray analysis. Human H460 lung cancer cells were cultured for 24 h in the presence or absence of 10 μM CTD; gene expression was then examined using microarray analysis. The results indicated that 8 genes were upregulated > 4-fold, 29 genes were upregulated >3–4-fold and 156 genes were upregulated >2–3-fold. In addition, 1 gene was downregulated >4 fold, 14 genes were downregulated >3–4-fold and 150 genes were downregulated >2–3 fold in H460 cells following exposure to CTD. It was found that CTD affected DNA damage genes, including DNIT3 and GADD45A, which were upregulated 2.26- and 2.60-fold, respectively, as well as DdiT4, which was downregulated 3.14-fold. In addition, the expression of genes associated with the cell cycle progression were altered, including CCND2, CDKL3 and RASA4, which were upregulated 2.72-, 2.19- and 2.72-fold, respectively; however, CDC42EP3 was downregulated 2.16-fold. Furthermore, apoptosis-associated genes were differentially expressed, including CARD6, which was upregulated 3.54-fold. In conclusion, the present study demonstrated that CTD affected the expression of genes associated with DNA damage, cell cycle progression and apoptotic cell death in human lung cancer H460 cells.

Benzyl isothiocyanate alters the gene expression with cell cycle regulation and cell death in human brain glioblastoma GBM 8401 cells

“…The isolated RNA was further conducted for cDNA synthesis, labeling and microarray hybridization. The fluorescent-labeled cDNA hybridization (Affymetrix GeneChip Human Gene 1.0 ST array; Affymetrix, Santa Clara, CA, USA) on the chip was conducted (18), and the fluorescence from each sample was measured by Asia BioInnovations Corporation (Taipei, Taiwan). Expression Console software (Affymetrix) with default RNA parameters (19,20) was used to analyze the data.…”

Cantharidin alters the expression of genes associated with the NKG2D-associated immune response in TSGH-8301 human bladder carcinoma cells

“…A previous study suggested that fibroblast growth factor receptor 2 gene mutations in bladder cancer may only serve a minor role in bladder carcinogenesis ( 31 ). Another previous study demonstrated that CTD alters gene expression in human lung cancer NCI-H460 cells in vitro ( 32 ). It has also been hypothesized that CTD alters the gene expression of bladder cancer cells; however, to the best of our knowledge this has not yet been tested.…”

Anticancer Attributes of Cantharidin: Involved Molecular Mechanisms and Pathways

“…Their work illustrated that CTD can generate both double-strand and single-strand DNA breaks in a leukemia cell line (CCRF-CEM) [ 14 ]. Afterward, the cytotoxic effects of CTD on a wide range of human cancer cell lines were examined by different researchers including skin cancer A431 [ 35 ], A375.S2 [ 36 ], bladder cancer T24, RT4 [ 37 ], non-small cell lung cancer (NSCLC) NCI-H460 [ 38 , 39 ], A549 [ 40 ], H358 [ 41 ], colorectal cancer colo 205 [ 42 ], hepatocellular carcinoma HepG2, Hep3B, gastric cancer BGC823, MGC803 [ 43 ], cholangiocarcinoma QBC939 [ 44 ], breast cancer cell MCF-7 [ 45 ], pancreatic cancer PANC-1, CFPAC-1 [ 46 , 47 ], oral carcinoma SAS, SCC-4, CAL-27 [ 48 ], TCA8113 [ 49 ], UMSSC [ 50 ] and ovarian cancer HO-8910PM [ 51 ], which intimate that it can effectively halt growth and proliferation of different human cancers ( Figure 3 ). Imatinib resistance is one of the great challenges for chronic myeloid leukemia (CML) therapy as 20–30% of patients remain resistant to it.…”

“…On the other hand, expression levels and cytoplasm to nucleus translocation of phosphorylated p53, MDC1 and phosphorylated H2A.X was increased [ 76 ]. In another study, cDNA microarray analysis revealed that CTD caused up-regulation of GADD45A (2.60-fold) and DNIT3 (2.26-fold) and down-regulation of DdiT4 (3.14-fold), which are DNA damage associated genes [ 38 ]. CTD caused sensitization of pancreatic cancer cells towards radiotherapy by elevating DNA damage and suppressing DNA repair associated genes namely UBE2T, RM1, RPA1, XRCC1, GTF2HH5, RAD51B, RAD50, RAD51B, PRKDC, LIG1, FANC1, DMC1, POLD3 and FAAP100 through JNK, ERK, p38, PKC and NF-κB pathways [ 78 ].…”