Apoptotic mechanism of human leukemia K562/A02 cells induced by magnetic iron oxide nanoparticles co-loaded with daunorubicin and 5-bromotetrandrin

Figures & data

Table 1 Summary of primer sequences for bcl-2, bax, caspase 3, survivin, and GAPDH

Gene	Primer for RT-PCR	Band size (bp)	Annealing temperature (°C)
bcl-2	Forward 5′-TGTGGCCTTCTTTGAGTTCG-3′	281	56
	Reverse 5′-TCACTTGTGGCCCAGATAGG-3′
bax	Forward 5′-TTTGCTTCAGGGTTTCATCC-3′	246	52
	Reverse 5′-CAGTTGAAGTTGCCGTCAGA-3′
caspase 3	Forward 5′-TTGATGCGTGATGTTTCTA-3′	223	51
	Reverse 5′-CAATGCCACAGTCCAGTTC-3′
survivin	Forward 5′-CTCAAGGACCACCGCATCTC-3′	392	55
	Reverse 5′-CCTCAATCCATGGCAGCCAG-3′
GAPDH	Forward 5′-GCCACATCGCTCAGACAC-3′	614	53
	Reverse 5′-CATCACGCCACAGTTTCC-3′

Abbreviation: RT-PCR, reverse transcriptase polymerase chain reaction.

Figure 1 The morphological characteristics of apoptosis in K562/A02 cells incubated for 48 hours. (DAPI staining, ×100). A) Control; B) DNR/Br Tet-Sol; C) DNR/Br Tet-MNPs. Arrows indicate cells with apoptotic nuclear condensation and fragmentation.

Abbreviations: DNR/Br Tet-Sol, daunorubicin and 5-bromotetrandrin in solution; DNR/Br Tet-MNPs, daunorubicin and 5-bromotetrandrin in magnetic nanoparticles.

Figure 2 The apoptotic rates of K562/A02 cells incubated for 48 hours by flow cytometry assay. A) Control; B) DNR-Sol; C) Br Tet-Sol; D) DNR/Br Tet-Sol; E) DNRMNPs; F) Br Tet- MNPs; G) DNR/Br Tet- MNPs; H) MNPs.

Abbreviations: DNR-Sol, daunorubicin in solution; Br Tet-Sol, 5-bromotetrandrin in solution; DNR/Br Tet-Sol, daunorubicin and 5-bromotetrandrin in solution; DNR-MNPs, daunorubicin in magnetic nanoparticles; Br Tet-MNPs, 5-bromotetrandrin in magnetic nanoparticles; DNR/Br Tet-MNPs, daunorubicin and 5-bromotetrandrin in magnetic nanoparticles; MNPs, magnetic nanoparticles.

Figure 3 Intracellular accumulation of DNR in K562/A02 cells incubated for 48 hours by flow cytometry assay. A) Intracellular fluorescence intensity associated with DNR in treated cells. a) Control, b) DNR-Sol, c) DNR/Br Tet-Sol, d) DNR-MNPs, e) DNR/Br Tet-MNPs; B) RFI of DNR in treated cells.

Abbreviations: DNR-Sol, daunorubicin in solution; DNR/Br Tet-Sol, daunorubicin and 5-bromotetrandrin in solution; DNR-MNPs, daunorubicin in magnetic nanoparticles; DNR/Br Tet-MNPs, daunorubicin and 5-bromotetrandrin in magnetic nanoparticles; FI, fluorescence intensity; RFI, relative fluorescence intensity.

Figure 4 Transcription of apoptosis-associated genes by RT-PCR analysis.

Note: Data are presented as means ± standard deviation (n = 3).

Abbreviations: DNR-Sol, daunorubicin in solution; DNR/Br Tet-Sol, daunorubicin and 5-bromotetrandrin in solution; DNR-MNPs, daunorubicin in magnetic nanoparticles; DNR/Br Tet-MNPs, daunorubicin and 5-bromotetrandrin in magnetic nanoparticles; MNPs, magnetic nanoparticles; RT-PCR, reverse transcriptase polymerase chain reaction.

Figure 5 Expression of apoptosis-associated proteins by Western blotting analysis.

Note: Data are presented as means ± standard deviation (n = 3).

Abbreviations: DNR-Sol, daunorubicin in solution; DNR/Br Tet-Sol, daunorubicin and 5-bromotetrandrin in solution; DNR-MNPs, daunorubicin in magnetic nanoparticles; DNR/Br Tet-MNPs, daunorubicin and 5-bromotetrandrin in magnetic nanoparticles; MNPs, magnetic nanoparticles.