Tamoxifen-loaded solid lipid nanoparticles-induced apoptosis in breast cancer cell lines

Figures & data

Table 1. The IC₅₀ of TAM, TAM-loaded SLN and SLN formulations on MCF-7 and MDA-MB231cells after 24, 48 and 72 h.

	IC50 (µg/mL)
Treatment	Cells	24 h	48 h	72 h
TAM	MCF-7 MDA-MB231	13.45 ± 0.46 17.21 ± 1.44	13.00 ± 0.98 16.87 ± 1.97	12.50 ± 0.91 15.97 ± 0.86
TAM-SLN	MCF-7 MDA-MB231	13.18^a ± 0.66 16.93 ± 0.82	12.50^a ± 1.50 16.00 ± 0.10	11.78^b ± 0.18 15.80 ± 0.69
SLN	MCF-7 MDA-MB231	287.5 ± 17.67 290.23 ± 1.4	262.05 ± 17.85 292.00 ± 12.48	222.33 ± 13.65 275.00 ± 1.00

Note: All values represent the mean ± standard deviation (n = 5).

^a,bMeans in each row with different superscripts are significantly different. TAM = tamoxifen; TAM-SLN = tamoxifen-loaded solid lipid nanoparticle and SLN = solid lipid nanaoparticles.

Figure 1. Phase contrast micrographs of MCF-7 cell treated with (A) vehicle (DMSO; control), (B) SLN (60 µg/mL) dispersed in DMSO, (C) TAM (12 µg/mL) dissolved in DMSO and (D) TAM-loaded SLN (12 µg/mL) dispersed in DMSO (magnification ×10).

Figure 2. Phase contrast micrographs of MDA-MB231 cell treated with (A) vehicle (DMSO; control), (B) SLN (80 µg/mL) dispersed in DMSO, (C) TAM (16 µg/mL) dissolved in DMSO and (D) TAM-loaded SLN (16 µg/mL) dispersed in DMSO (magnification ×10).

Figure 3. Fluorescence microscopy of MCF-7 cells treated with (A) vehicle, (DMSO; control), TAM (B) and TAM-loaded SLN (C). Cell shrinkage (thin arrow) and apoptotic cells are evident (thick arrow) (magnification ×40).

Figure 4. Fluorescence microscopy of MDA-MB231 cells treated with (A) vehicle, (DMSO; control), TAM (B) and TAM-loaded SLN (C). Cell shrinkage (thin arrow) and apoptotic cells (thick arrow) are evident (magnification ×40).

Table 2. Effect of TAM and TAM-loaded SLN at concentration 12 µg/mL on MCF-7 cell cycle distribution.

	MCF-7 cells (%)
Treatment	Time (h)	G0/G1 R3	S R4	G2/M R5	Sub-G0/G1 R2
Control	24	59.19^a ± 2.0	13.34 ± 1.18	27.06 ± 2.50	1.53^a ± 0.20
	48	58.39^a ± 5.0	12.35 ± 0.56	28.61 ± 0.54	1.59^a ± 0.52
	72	55.49^a ± 0.5	11.40 ± 0.53	29.48 ± 0.50	4.40^b ± 0.60
TAM	24	49.00^b ± 0.77	5.10 ± 0.60	4.48 ± 0.24	42.5^c ± 1.25
	48	46.30^b ± 1.00	11.72 ± 0.75	5.34 ± 0.62	37.44^d ± 1.00
	72	49.85^b ± 1	3.56 ± 0.24	3.97 ± 0.66	43.53^c ± 0.87
TAM-SLN	24	54.78^a ± 1	2.731 ± 0.53	3.87 ± 0.53	39.0^d ± 1.33
	48	45.32^b ± 1.14	11.04 ± 0.96	5.13 ± 1.02	38.15^d ± 1.05
	72	47.18^b ± 3.57	1.65 ± 0.10	1.42 ± 0.13	50.41^e ± 3.77

Note: All values are expressed as mean ± standard deviation (n = 3).

^a,b,c,d,eMeans in each column with different superscripts are significantly different (p < 0.05).

Table 3. Effect of TAM and TAM-loaded SLN at concentration 16 µg/mL on MDA-MB231 cell cycle distribution.

	MDA-MB-231 cells (%)
Treatment	Time (h)		G0/G1 R3	S R4	G2/M R5	Sub-G0/G1 R2
Control	24	53.99^a ± 0.62		16.60 ± 0.66	23.75 ± 0.82	5.93^a ± 0.62
	48	54.00^a ± 0.10		14.02 ± 0.31	28.54 ± 0.53	4.85^a ± 0.27
	72	67.23^b ± 0.23		10.54 ± 0.29	16.21 ± 0.29	7.40^a ± 0.33
TAM	24	61.87^b ± 0.83		7.55 ± 0.31	16.81 ± 0.47	14.52^b ± 0.85
	48	53.58^a ± 0.80		3.27 ± 0.17	9.96 ± 0.17	33.63^c ± 0.42
	72	19.54^c ± 1		5.17 ± 0.24	6.30 ± 0.26	69.71^d ± 0.51
TAM-SLN	24	62.07^b ± 7.06		8.47 ± 2.64	15.15 ± 1.78	14.60^b ± 2.61
	48	35.88^d ± 0.70		15.13 ± 1.45	27.82 ± 0.75	19.34^e ± 0.45
	72	25.05^c ± 0.5		2.33 ± 0.17	9.37 ± 0.37	63.85^f ± 0.33

Note: All values are expressed as mean ± standard deviation (n = 3).

^a,b,c,d,e,fMeans in each column with different superscripts are significantly different (p < 0.05).

Table 4. Effect of TAM and TAM-loaded SLN at concentration 15 µg/mL on MCF-7 cell cycle distribution.

	MCF-7 cells (%)
Treatment	Time (h)	G0/G1 R3	S R4	G2/M R5	Sub-G0/G1 R2
Control	24	59.19^a ± 2.0	13.34 ± 1.18	27.06 ± 2.5	1.53^a ± 0.20
TAM	24	13.47^b ± 1.5	5.97 ± 1.81	4.47 ± 0.84	76.30^b ± 4.0
TAM-SLN	24	7.71^c ± 2.06	3.05 ± 1.00	2.30 ± 0.60	86.92^c ± 3.58

Note: All values are expressed as mean ± standard deviation (n = 3).

^a,bMeans in each column with different superscripts are significantly different (p < 0.05).

Table 5. Effect of TAM and TAM-loaded SLN at concentration 18 µg/mL on MDA-MB231 cell cycle distribution.

	MDA-MB231 cells (%)
Treatment	Time (h)	G0/G1 R3	S R4	G2/M R5	Sub-G0/G1 R2
Control	24	53.99^a ± 0.62	16.60 ± 0.66	23.75 ± 0.82	5.93^a ± 0.62
	48	54.00^a ± 0.10	14.02 ± 0.31	28.54 ± 0.53	4.85^a ± 0.27
TAM	24	32.86^b ± 3.15	8.73 ± 0.23	20.63 ± 0.34	35.22^b ± 2.80
	48	10.86^c ± 0.33	0.92 ± 0.42	1.10 ± 0.12	87.90^d ± 0.40
TAM-SLN	24	36.90^b ± 0.73	10.13 ± 0.17	26.05 ± 0.50	22.59^c ± 0.65
	48	10.25^c ± 0.18	1.67 ± 0.08	2.47 ± 0.1	85.94^d ± 0.26

Note: All values are expressed as mean ± standard deviation (n = 3).

^a,b,c,dMeans in each column with different superscripts are significantly different (p < 0.05).

Table 6. Effect of TAM and TAM-loaded SLN at concentration 20 µg/mL on MDA-MB231 cell cycle distribution.

	MDA-MB231 cells (%)
Treatment	Time (h)	G0/G1 R3	S R4	G2/M R5	Sub-G0/G1 R2
Control	24	53.99^a ± 0.62	16.60 ± 0.66	23.75 ± 0.82	5.93^a ± 0.62
	48	54.00^a ± 0.10	14.02 ± 0.31	28.54 ± 0.53	4.85^a ± 0.27
TAM	24	14.11^b ± 1.26	1.57 ± 0.61	2.53 ± 0.67	82.06^b ± 2.90
	48	2.03^c ± 0.15	0.10 ± 0.02	0.13 ± 0.08	97.88^c ± 0.17
TAM-SLN	24	32.97^d ± 1.46	5.13 ± 1.23	14.23 ± 1.1	47.64^d ± 3.74
	48	6.34^e ± 0.73	0.39 ± 0.04	0.53 ± 0.06	93.05^e ± 0.72

Note: All values are expressed as mean ± standard deviation (n = 3).

^a,b,c,d,eMeans in each column with different superscripts are significantly different (p < 0.05).