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Differential migration-related gene expression and altered cytokine secretion in response to serum starvation in cultured MDA-MB-231 cells

Naghmeh Ahmadiankia
Naghmeh Ahmadiankia
, 
Mehdi Bagheri
Mehdi Bagheri
 und 
Mozhgan Fazli
Mozhgan Fazli
   | 31. März 2020
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Article Category: Original article
Online veröffentlicht: 31. März 2020
Seitenbereich: 123 - 129
DOI: https://doi.org/10.1515/abm-2019-0051
Schlüsselwörter
© 2019 Naghmeh Ahmadiankia et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

Serum starvation inhibited cell migration. (A) Cells migrated to the basal side of the membrane were stained and images were captured at a magnification of 100×. (B) Results are presented in graphs as the average number of migrated cells on the underside of the filter. **donates P < 0.01, as compared to the control. Experiments were performed in triplicate wells for each condition and repeated at least twice. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.
Serum starvation inhibited cell migration. (A) Cells migrated to the basal side of the membrane were stained and images were captured at a magnification of 100×. (B) Results are presented in graphs as the average number of migrated cells on the underside of the filter. **donates P < 0.01, as compared to the control. Experiments were performed in triplicate wells for each condition and repeated at least twice. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.

Figure 2

Expression of migration-related genes in MDA-MB-231 cells were downregulated by serum starvation after 24, 48, and 72 h. Relative levels are presented as fold change compared with untreated. * and ** denote P < 0.05 and P < 0.01, respectively, as compared to the control. All experiments were performed in triplicate wells for each condition and repeated at least twice. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.
Expression of migration-related genes in MDA-MB-231 cells were downregulated by serum starvation after 24, 48, and 72 h. Relative levels are presented as fold change compared with untreated. * and ** denote P < 0.05 and P < 0.01, respectively, as compared to the control. All experiments were performed in triplicate wells for each condition and repeated at least twice. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.

Figure 3

Cytokine release was assessed by ELISA. TGFβ1, MMP9, IL8, and NO concentrations (mean pg/mL ± SD) were measured in the super-natant of MDA-MB-231 cells underwent serum starvation for 24, 48, and 72 h. Significance was set at *P < 0.05 and **P < 0.01. Experiments were performed in triplicate. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.
Cytokine release was assessed by ELISA. TGFβ1, MMP9, IL8, and NO concentrations (mean pg/mL ± SD) were measured in the super-natant of MDA-MB-231 cells underwent serum starvation for 24, 48, and 72 h. Significance was set at *P < 0.05 and **P < 0.01. Experiments were performed in triplicate. The significance of difference between the groups was assessed by one-way ANOVA followed by Dunnett’s test.

Real-time PCR primer sequences

PrimersSequenceProduct size
β-cateninF: AAAATGGCAGTGCGTTTAG100 bp
R: TTTGAAGGCAGTCTGTCGTA
TwistF: TGCGGAAGATCATCCCCACG137 bp
R: GCTGCAGCTTGCCATCTTGGA
ZEB1F: TGCACTGAGTGTGGAAAAGC237 bp
R: TGGTGATGCTGAAAGAGACG
VimentinF: ACCCGCACCAACGAGAAGGT90 bp
R: ATTCTGCTGCTCCAGGAAGCG
FibronectinF: TCCTTGCTGGTATCATGGCAG74 bp
R: AGACCCAGGCTTCTCATACTTGA
ICAM1F: AGGCCACCCCAGAGGACAAC406 bp
R: CCCATTATGACTGCGGCTGCTA
VEGFF: CCTTGCTGCTCTACCTCCAC280 bp
R: ATCTGCATGGTGATGTTGGA
GAPDHF: AAGGTGAAGGTCGGAGTCAAC102 bp
R: GGGGTCATTGATGGCAACAATA
eISSN:
1875-855X
Sprache:
Englisch
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Fachgebiete der Zeitschrift:
Medizin, Gesundheitsfachberufe, Vorklinische Medizin, Grundlagenmedizin, andere, Klinische Medizin
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