Apoptosis of human colon carcinoma HT-29 cells induced by ceramide

Table 1 Primer sequences of Bcl-2 family gene members

Gene	Primer sequence	Size (bp)	Temperature (°C)
Bax	Sense: 5’-CCAGCTGCCTTGGACTGT-3’	135	61
	Antisense: 5’-ACCCCCTCAAGACCACTCTT-3’
Bcl-xl	Sense: 5’-GTAAACTGGGGTCGCATTGT-3’	146	60
	Antisense: 5’-TGGATCCAAGGCTCTAGGTG-3’
Bcl-2	Sense: 5’-GGCAATGTGACTTTTTCCAA-3’	137	55
	Antisense: 5’-GGCTGATATTCTGCAACACTG-3’
Bad	Sense: 5’-AGGGCTGACCCAGATTCC-3’	178	60
	Antisense: 5’-GTGACGCAACGGTTAAACCT-3’
Bid	Sense: 5’-GCTTCCAGTGTAGACGGAGC-3’	116	60
	Antisense: 5’-GTGCAGATTCATGTGTGGATG-3’
β-actin	Sense: 5’-GTGGACATCCGCAAAGAC-3’	303	58
	Antisense: 5’-TCAACGCAATGTGGGAAG-3’

Table 2 Apoptosis rate of HT-29 cells induced by C₂-ceramide (mean ± SD, %)

C2-ceramide	Cell count (n)	12 h	24 h
0 μmol/L	300	5.33 ± 1.53	8.67 ± 2.52
12.5 μmol/L	300	14.7 ± 2.52a	28.0 ± 9.85a
25 μmol/L	300	33.3 ± 8.37a	43.3 ± 12.3a
50 μmol/L	300	64.7 ± 7.51a	81.3 ± 9.07a

^aP < 0.05 vs control.

Table 3 C₂-ceramide damaged mitochondria function of HT-29 cells (mean ± SD)

C2-ceramide (mmol/L)	1 h	3 h	6 h	9 h	12 h	24 h
0	0.505 ± 0.061	0.596 ± 0.050	0.528 ± 0.083	1.088 ± 0.134	1.371 ± 0.066	1.770 ± 0.161
12.5	0.478 ± 0.044	0.609 ± 0.048	0.502 ± 0.029	0.833 ± 0.149a	1.036 ± 0.122	1.560 ± 0.220
25	0.487 ± 0.051	0.568 ± 0.048	0.404 ± 0.033a	0.646 ± 0.158a	0.760 ± 0.086a	1.484 ± 0.346a
50	0.490 ± 0.054	0.541 ± 0.049	0.345 ± 0.050a	0.456 ± 0.071a	0.708 ± 0.073a	1.342 ± 0.061a

^aP < 0.05 vs control.