Antiproliferation effect of imatinib mesylate on MCF7, T-47D tumorigenic and MCF 10A nontumorigenic breast cell lines via PDGFR-β, PDGF-BB, c-Kit and SCF genes

Figures & data

Figure 1 QuantiTect Primer Assay specification.

Abbreviations: PDGFR, platelet-derived growth factor receptor; PDGF-BB, platelet-derived growth factor BB; SCF, stem cell factor.

Figure 2 Time- and dose-dependent antiproliferation effect of imatinib mesylate with clinically relevant concentrations (2–10 µM) in two MCF7 and T-47D breast tumorigenic cell lines and a nontumorigenic breast cell line MCF 10A.

Notes: Antiproliferation effect is reported as a reverse percentage of viable cells relative to untreated control cells. P<0.05 was considered as significant.

Figure 3 The concentration of imatinib mesylate that is required for 50% inhibition of two MCF7 and T-47D breast tumorigenic cell lines and a nontumorigenic breast cell line MCF 10A proliferation (half minimal inhibitory concentration) after 144 h treatment compared to untreated cell lines.

Note: *P<0.05 was considered as significant.

Figure 4 Apoptosis terminal deoxynucleotidyl transferase dUTP nick end labeling assay detection of time-dependent imatinib mesylate apoptosis induction within 48–144 h treatment with corresponding half minimal inhibitory concentration of imatinib mesylate for each cell line.

Notes: Apoptosis induction effect is reported as a percentage of apoptotic cells relative to untreated control. *P<0.05 was considered as significant.

Figure 5 The effect of imatinib mesylate on cell cycle distribution of MCF7, T-47D tumorigenic and MCF 10A nontumorigenic breast cell lines.

Notes: (A, D and G) Untreated control MCF 10A, MCF7 and T-47D cell lines, respectively, after 144 h. Treated MCF 10A, MCF7 and T-47D cell lines, respectively, with each cell line individual half maximal inhibitory concentration of imatinib mesylate for (B, E and H) 72 and (C, F and I) 144 h, flow cytometry analysis was carried out on treated and untreated cell lines. (J) The quantitative analysis shows the cell cycle arrest at G₂/M phase. Data are shown as the mean ± standard deviation (n=3). *Significant difference (P<0.05) compared with untreated control.
Abbreviations: G, gap; M, mitosis; S, synthesis; PI-A, propidium iodide-A.

Figure 6 Comparative expression of the PDGFR-β, c-Kit, PDGF-BB and SCF genes compared to house-keeping gene (β-actin) in untreated two MCF7 and T-47D breast tumorigenic cell lines and a nontumorigenic breast cell line MCF 10A.

Abbreviations: PDGF-BB, platelet-derived growth factor BB; PDGFR, platelet-derived growth factor receptor; SCF, stem cell factor.

Figure 7 The relative gene expression pattern profiling of the PDGFR-β, c-Kit, PDGF-BB and SCF genes in imatinib mesylate-treated cell lines compared to untreated controls and normalized with HK genes.

Notes: (A) Boxes represents the interquartile range, or the middle 50% of observations. The dotted line represents the median gene expression. Whiskers represent the minimum and maximum observations. (B) Calculated gene expression change using 2^−ddCT method. *P<0.05 was considered as significant.
Abbreviations: PDGF-BB, platelet-derived growth factor BB; PDGFR, platelet-derived growth factor receptor; SCF, stem cell factor.

Table 1 The relative gene expression analysis report

Gene	Type	Expression	Standard error	P (H1)	Result
MCF 10A
β-Actin	REF	1.000
PDGFR-β	TRG	0.837	0.807–0.872	0.023	Down
c-Kit	TRG	1.043	0.996–1.129	0.198
PDGF-BB	TRG	1.135	1.067–1.199	0.028	Up
SCF	TRG	1.065	0.979–1.151	0.293
MCF7
β-A ctin	REF	1.000
PDGFR-β	TRG	0.502	0.465–0.546	0.010	Down
c-Kit	TRG	0.771	0.710–0.851	0.010	Down
PDGF-BB	TRG	0.463	0.429–0.524	0.020	Down
SCF	TRG	1.133	1.036–1.209	0.012	Up
T-47D
β-Actin	REF	1.000
PDGFR-β	TRG	0.414	0.379–0.463	0.028	Down
PDGF-BB	TRG	0.315	0.290–0.343	0.028	Down
SCF	TRG	1.785	1.646–1.953	0.000	Up

Abbreviations: P (H1), probability of alternate hypothesis that difference between sample and control groups is due only to chance (P-value); PDGF-BB, platelet-derived growth factor BB; PDGFR, platelet-derived growth factor receptor; REF, reference endogenous genes; SCF, stem cell factor; TRG, target endogenous genes.

Figure 8 Relative expression of the PDGFR-β, c-Kit, PDGF-BB and SCF proteins compared to house-keeping protein (β-actin) in untreated cell lines.

Abbreviations: PDGF-BB, platelet-derived growth factor BB; PDGFR, platelet-derived growth factor receptor; SCF, stem cell factor.

Figure 9 The effect of imatinib mesylate treatment on PDGFR-β, c-Kit, PDGF-BB and SCF protein expression related to untreated cell lines.

Notes: (A) Western blotting assay. (B) Protein expression pattern profiling. *P<0.05 was considered as significant.
Abbreviations: PDGF-BB, platelet-derived growth factor BB; PDGFR, platelet-derived growth factor receptor; Tr, treated-cell lines; UnTr, untreated-cell lines.