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Cisplatin sensitivity is enhanced in non-small cell lung cancer cells by regulating epithelial-mesenchymal transition through inhibition of eukaryotic translation initiation factor 5A2.

Xu G, Yu H, Shi X, Sun L, Zhou Q, Zheng D, Shi H, Li N, Zhang X, Shao G - BMC Pulm Med (2014)

Bottom Line: Recent studies have suggested eIF5A-2 may function as a proliferation-related oncogene in tumorigenic processes.We found low concentrations of GC7 have little effect on NSCLC viability, but could enhance cisplatin cytotoxicity in NSCLC cells.GC7 also could reverse mesenchymal phenotype in NCI-H1299 and prevented A549 cells undergoing EMT after TGF-β1 inducement. eIF5A-2 knockdown resulted in EMT inhibition.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic & Cardiovascular Surgery, Lihuili Hospital, Ningbo Medical Center, Affiliated Hospital of Medical School of Ningbo University, NO 57 Xingning Road, Ningbo 315041, China. guofengshaolihuili@163.com.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) has been believed to be related with chemotherapy resistance in non-small cell lung cancer (NSCLC). Recent studies have suggested eIF5A-2 may function as a proliferation-related oncogene in tumorigenic processes.

Methods: We used cell viability assays, western blotting, immunofluorescence, transwell-matrigel invasion assay, wound-healing assay combined with GC7 (a novel eIF5A-2 inhibitor) treatment or siRNA interference to investigate the role of eIF5A-2 playing in NSCLC chemotherapy.

Results: We found low concentrations of GC7 have little effect on NSCLC viability, but could enhance cisplatin cytotoxicity in NSCLC cells. GC7 also could reverse mesenchymal phenotype in NCI-H1299 and prevented A549 cells undergoing EMT after TGF-β1 inducement. eIF5A-2 knockdown resulted in EMT inhibition.

Conclusion: Our data indicated GC7 enhances cisplatin cytotoxicity and prevents the EMT in NSCLC cells by inhibiting eIF5A-2.

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GC7 enhanced cisplatin sensitivity in NSCLC cells via inhibition of eIF5A-2. (A) eIF5A-2 siRNA inhibits eIF5A-2 in both A549 and NCI-H1299 cells. (B–C) Comparing changes in cisplatin sensitivity in A549 and NCI-H1299 NSCLC cells after treatment with eIF5A-2 siRNA alone or combined with GC7.
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Fig3: GC7 enhanced cisplatin sensitivity in NSCLC cells via inhibition of eIF5A-2. (A) eIF5A-2 siRNA inhibits eIF5A-2 in both A549 and NCI-H1299 cells. (B–C) Comparing changes in cisplatin sensitivity in A549 and NCI-H1299 NSCLC cells after treatment with eIF5A-2 siRNA alone or combined with GC7.

Mentions: GC7 can inhibit the activity of eIF5A-2 (In Additional file 2). In order to discover the mechanism by which GC7 enhanced cisplatin sensitivity, we transfected eIF5A-2 siRNA into A549 and NCI-H1299 cells to interfere with eIF5A-2 expression, and found that eIF5A-2 expression was significantly inhibited in both NSCLC cell lines (Figure 3A). We then treated these transfected cells with cisplatin alone, or cisplatin combined with GC7, and carried out CCK-8 cell viability assays. Without GC7, NCI-H1299 cells were the most sensitive to cisplatin after eIF5A-2 siRNA transfection: the IC50 at 72 h was 4.468 μg/mL (4.093–4.842 μg/mL; Table 2). Although A549 cells remained sensitive to cisplatin, the IC50 value was lower: 2.626 μg/mL (2.466–2.785 μg/mL; P = 0.0145 vs. cisplatin alone. Table 2). In contrast, when cisplatin treatment was combined with GC7 after eIF5A-2 siRNA transfection, there was little change in the cisplatin sensitivity of both cell lines: the IC50 values at 72 h were 3.982 μg/mL (3.609–4.356 μg/mL; P = 0.0648) and 2.434 μg/mL (2.307–2.560 μg/mL; P =0.0571) in NCI-H1299 and A549 cells, respectively (Table 2; Figure 3B,C). As GC7 also inhibits eIF5A-1’s activity, we evaluated the role of eIF5A-1 in this process. Western Blot analysis indicated that eIF5A-1 was expressed in the control cells of both cell lines; however the expression of eIF5A-1 was higher in NCI-H1299 cells compared to A549 cells. Moreover, We also evaluated the effect of eIF5A-1 in the siRNA transfected cell. The results showed that when cisplatin treatment was combined with GC7 after eIF5A-1 siRNA transfection, there was little change in the cisplatin sensitivity of both cell lines (In Additional file 3).Figure 3


Cisplatin sensitivity is enhanced in non-small cell lung cancer cells by regulating epithelial-mesenchymal transition through inhibition of eukaryotic translation initiation factor 5A2.

Xu G, Yu H, Shi X, Sun L, Zhou Q, Zheng D, Shi H, Li N, Zhang X, Shao G - BMC Pulm Med (2014)

GC7 enhanced cisplatin sensitivity in NSCLC cells via inhibition of eIF5A-2. (A) eIF5A-2 siRNA inhibits eIF5A-2 in both A549 and NCI-H1299 cells. (B–C) Comparing changes in cisplatin sensitivity in A549 and NCI-H1299 NSCLC cells after treatment with eIF5A-2 siRNA alone or combined with GC7.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4232729&req=5

Fig3: GC7 enhanced cisplatin sensitivity in NSCLC cells via inhibition of eIF5A-2. (A) eIF5A-2 siRNA inhibits eIF5A-2 in both A549 and NCI-H1299 cells. (B–C) Comparing changes in cisplatin sensitivity in A549 and NCI-H1299 NSCLC cells after treatment with eIF5A-2 siRNA alone or combined with GC7.
Mentions: GC7 can inhibit the activity of eIF5A-2 (In Additional file 2). In order to discover the mechanism by which GC7 enhanced cisplatin sensitivity, we transfected eIF5A-2 siRNA into A549 and NCI-H1299 cells to interfere with eIF5A-2 expression, and found that eIF5A-2 expression was significantly inhibited in both NSCLC cell lines (Figure 3A). We then treated these transfected cells with cisplatin alone, or cisplatin combined with GC7, and carried out CCK-8 cell viability assays. Without GC7, NCI-H1299 cells were the most sensitive to cisplatin after eIF5A-2 siRNA transfection: the IC50 at 72 h was 4.468 μg/mL (4.093–4.842 μg/mL; Table 2). Although A549 cells remained sensitive to cisplatin, the IC50 value was lower: 2.626 μg/mL (2.466–2.785 μg/mL; P = 0.0145 vs. cisplatin alone. Table 2). In contrast, when cisplatin treatment was combined with GC7 after eIF5A-2 siRNA transfection, there was little change in the cisplatin sensitivity of both cell lines: the IC50 values at 72 h were 3.982 μg/mL (3.609–4.356 μg/mL; P = 0.0648) and 2.434 μg/mL (2.307–2.560 μg/mL; P =0.0571) in NCI-H1299 and A549 cells, respectively (Table 2; Figure 3B,C). As GC7 also inhibits eIF5A-1’s activity, we evaluated the role of eIF5A-1 in this process. Western Blot analysis indicated that eIF5A-1 was expressed in the control cells of both cell lines; however the expression of eIF5A-1 was higher in NCI-H1299 cells compared to A549 cells. Moreover, We also evaluated the effect of eIF5A-1 in the siRNA transfected cell. The results showed that when cisplatin treatment was combined with GC7 after eIF5A-1 siRNA transfection, there was little change in the cisplatin sensitivity of both cell lines (In Additional file 3).Figure 3

Bottom Line: Recent studies have suggested eIF5A-2 may function as a proliferation-related oncogene in tumorigenic processes.We found low concentrations of GC7 have little effect on NSCLC viability, but could enhance cisplatin cytotoxicity in NSCLC cells.GC7 also could reverse mesenchymal phenotype in NCI-H1299 and prevented A549 cells undergoing EMT after TGF-β1 inducement. eIF5A-2 knockdown resulted in EMT inhibition.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic & Cardiovascular Surgery, Lihuili Hospital, Ningbo Medical Center, Affiliated Hospital of Medical School of Ningbo University, NO 57 Xingning Road, Ningbo 315041, China. guofengshaolihuili@163.com.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) has been believed to be related with chemotherapy resistance in non-small cell lung cancer (NSCLC). Recent studies have suggested eIF5A-2 may function as a proliferation-related oncogene in tumorigenic processes.

Methods: We used cell viability assays, western blotting, immunofluorescence, transwell-matrigel invasion assay, wound-healing assay combined with GC7 (a novel eIF5A-2 inhibitor) treatment or siRNA interference to investigate the role of eIF5A-2 playing in NSCLC chemotherapy.

Results: We found low concentrations of GC7 have little effect on NSCLC viability, but could enhance cisplatin cytotoxicity in NSCLC cells. GC7 also could reverse mesenchymal phenotype in NCI-H1299 and prevented A549 cells undergoing EMT after TGF-β1 inducement. eIF5A-2 knockdown resulted in EMT inhibition.

Conclusion: Our data indicated GC7 enhances cisplatin cytotoxicity and prevents the EMT in NSCLC cells by inhibiting eIF5A-2.

Show MeSH
Related in: MedlinePlus