H9, a novel herbal extract, demonstrated cytotoxicity in A549 non-small cell lung cancer (NSCLC) cell lines. In this study, we investigated whether H9, and/or co-treatment with an anticancer drug, pemetrexed (PEM), inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. The mice ...
H9, a novel herbal extract, demonstrated cytotoxicity in A549 non-small cell lung cancer (NSCLC) cell lines. In this study, we investigated whether H9, and/or co-treatment with an anticancer drug, pemetrexed (PEM), inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. The mice were separated into groups and administered H9 and PEM for 2 weeks. Protein and mRNA levels were detected using western blotting and reverse transcription polymerase chain reaction, respectively; immunohistochemistry (IHC) was also performed on the tumor tissues. H9 and co-treatment with PEM induced the cleavage of proapoptotic factors, such as caspase-3, caspase-8, caspase-9, and poly(ADP)-ribose polymerase (PARP). Expression levels of cell-death receptors involving Fas/FasL, TNF-related apoptosisinducing ligands (TRAIL), and TRAIL receptors were increased by H9 and co-treatment with PEM. Furthermore, analysis of levels of cell-cycle modulating proteins indicated that tumor cells were arrested in the G1/S phase. In addition, the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt survival signaling pathways were inhibited by H9 and co-treatment with PEM. In conclusion, H9 and co-treatment with PEM inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. These results indicate that H9 and co-treatment with PEM can be used as an anticancer therapy in NSCLC.
H9, a novel herbal extract, demonstrated cytotoxicity in A549 non-small cell lung cancer (NSCLC) cell lines. In this study, we investigated whether H9, and/or co-treatment with an anticancer drug, pemetrexed (PEM), inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. The mice were separated into groups and administered H9 and PEM for 2 weeks. Protein and mRNA levels were detected using western blotting and reverse transcription polymerase chain reaction, respectively; immunohistochemistry (IHC) was also performed on the tumor tissues. H9 and co-treatment with PEM induced the cleavage of proapoptotic factors, such as caspase-3, caspase-8, caspase-9, and poly(ADP)-ribose polymerase (PARP). Expression levels of cell-death receptors involving Fas/FasL, TNF-related apoptosisinducing ligands (TRAIL), and TRAIL receptors were increased by H9 and co-treatment with PEM. Furthermore, analysis of levels of cell-cycle modulating proteins indicated that tumor cells were arrested in the G1/S phase. In addition, the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt survival signaling pathways were inhibited by H9 and co-treatment with PEM. In conclusion, H9 and co-treatment with PEM inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. These results indicate that H9 and co-treatment with PEM can be used as an anticancer therapy in NSCLC.
Oligo(dT)-primed RNA (5 µg) was reverse transcribed with M-MuLV reverse transcriptase (New England Biolabs, Ipswich, MA, USA). A reverse transcription polymerase chain reaction (RT-PCR) analysis was performed using a PCR thermal cycler Dice instrument (TaKaRa, Otsu, Shiga, Japan) with the following primer sets: Fas: 5’-AGG GAT TGG AAT TGA GGA AG-3’ (forward), 5’-ATG GGC TTT GTC TGT GTA CT-3’ (reverse); FasL: 5’-AGT CCA CCC CCT GAA AAA AA-3’ (forward), 5’-ATT CCA TAG GTG TCT TCC CA-3’ (reverse); GAPDH: 5’-GGC TGC TTT TAA CTC TGG TA-3’ (forward), 5’-TGG AAG ATG GTG ATG GGA TT-3’ (reverse); TRAIL: 5’-GTC TCT CTG TGT GGC TGT AA-3’ (forward), 5’-TGT TGC TTC TTC CTC TGG CT-3’ (reverse); TRAIL Receptor: 5’-AAG TTT GTC GTC GTC GGG GT-3’ (forward), 5’-TGG TGC AGG GAC TTC TCT CT-3’ (reverse); CIAP-1: 5’-CAG GTC CCT CGT ATC AAA AC-3’ (forward), 5’-TAA AAA CCA GCA CGA GCA AG-3’ (reverse); and cFLIPL: 5’-CGA GCA CCG AGA CTA CGA CA-3’ (forward), 5’-GCC CTC TGA CAC CAC ATA GT-3’ (reverse).
대상 데이터
Anti-rabbit and anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Millipore (Billerica, MA, USA). Antibodies against PI3K, Akt, p27, p21, cyclin D1, cyclin A, cyclin E, and GAPDH were purchased from Santa Cruz Biotechnology.
이론/모형
After 20 h of growth, the cells were treated with various concentrations of H9, PEM, and cotreatment for 24 h. The cell viability was estimated using the trypan blue assay. Briefly, the medium was removed and washed using PBS, harvested using trypsin-EDTA, and then centrifuged at 100 ×g for 5 min at 4℃.
성능/효과
H&E staining revealed that there were no differences between the groups, demonstrating that the H9, PEM, and combined treatments did not induce tissue damage.
H9 and treatments combined with PEM did not alter the Bax expression, whereas the FLIPL, cIAP-1, Bcl-2, STAT3, and Bcl-xL expression levels were inhibited (Fig
We therefore measured the body weight of the mice every week. No differences in body weights were observed between the control and drug-treated groups (Fig. 3A), suggesting that the H9, PEM, and combined treatments were non-toxic.
We measured the tumor volume twice a week. The H9 and co-treatment groups showed that the H9-treated group efficiently suppressed tumor growth compared with the other groups (Fig. 2).
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