Mutation analysis of APC gene in gastric cancer with microsatellite instability

Table 1 Primer Sequences used in APC mutation analysis

Exon	GC Clamp	Primer Sequences	Size (bp)
15.1	2/0	F-TTCAGGCAAATCCTAAGAGA/R-TTGAGCCAGGAGACATAATA	329
15.2	0/2	F-GGAATCTCATGGCAAATAGG/R-TCATCATGTCGATTGGTGTC	284
15.3	2/0	F-AGCAAAGTCTCTATGGTGAT/R-TGACACTTCTTCCATGACTT	387
15.4	0/2	F-CTACCATCCAGCAACAGAAA/R-TGGCATAAGGCATAGAACAT	312
15.5	2/0	F-AAGCTCTGCTGCCCATACAC/R-CTAGGTCGGCTGGGTATTGA	282
15.6	2/5	F-CTTATGCCAAATTAGAATAC/R-ACTGCTCATCTGAATATTTA	306
15.7	7/5	F-ATGATGGAGAACTAGATACA/R-ATCAGTGCTCTCAGTATAAA	253
15.8	2/5	F-AAGACCCAAACACATAATAG/R-TAATTGGTAGGCTTATCATC	327
15.9	2/0	F-AAATCGAGTGGGTTCTAATC/R-TTCTCACTGCTTGAAGACAT	355
15.10	2/6	F-CAGCCTATTGATTATAGTTT/R-AACATATTGGAGTATCTTCT	344
15.11	2/0	F-TTAACCAAGAAACAATACAG/R-CTTCGCTCACAGGATCTT	320
15.12	2/0	F-GGAAGCAGATTCTGCTAATA/R-ATAGTGTTCAGGTGGACTTT	287
15.13	2/5	F-AGAATCAGCCAGGCACAAAG/R-CATGGTTTGTCCAGGGCTAT	310
15.14	2/6	F-TGGTAAGTGGCATTATAAGC/R-AGTATCAGCATCTGGAAGAA	279
15.15	5/2	F-AAACCAAGCCAGAAGTACCT/R-AGGCTGCTCTGATTCTGTTT	335
15.16	2/5	F-CGATGAGCCATTTATACA/R-TTTGCTTTACGTGATGAC	291
15.17	4/2	F-ATTATTTCTGCCATGCCAAC/R-TTCGATTGTTAGATCACTTA	300
15.18	2/6	F-TGCCACGGGTGTATTGTGTT/R-TTTCCCTTTGGGCATAGCAG	351
15.19	4/2	F-GGTGATATTCTTGCAGAATG/R-CACGTGTCCTATATTCAGTA	250
15.20	2/0	F-GAAACCAACTTCACCAGTAA/R-AATAGGCGTGTAATGATGAG	378
15.21	2/0	F-AGCTCCCAAATAATGAAGAT/R-GATTGTTGGTTGGAGGTTAG	342
15.22	2/6	F-GCTAAAGTTACCAGCCACAC/R-TTGGTCAATGTCACTGAGAG
15.23	2/0	F-CCAGTCATCCAAAGACATAC/R-CAACAGGTCATCTTCAGAGT
15.24	2/5	F-TCACAGGGAGAACCAAGTAA/R-TTTGCACCTTCCTGAATAGC
15.25	2/3	F-ATGGGTGGCATATTAGGTGA/R-TCGTGGGCCTTTATTACTTG

GC clamp: 1 = CGCCCGCCGCGCCCCGCGCCCGTCCCGCCGCCCCCGCCCG; 2 = CGCCCGCCGCGCCCCGCGCCCGGCCCGCCGCCCCCGCCCG; 3 = CG; 4 = GCGCG; 5 = CGTCCCGC; 6 = GCGCCCGTCCCGCCC; 7 = CGCCCGCCGCGCCCCGCGCCCGTCCCGCCGCCCCCGCCCGGCCCG

Table 2 APC mutation in 12 gastric carcinomas

Exon region	Nucleotide alteration	Nucleotide location	Codon number
15.4	C→A	2883	940
15.5	insA	2866	935
15.10	del A	3835 or 3836	1258
15.11	del AAAA	3981	1307
15.12	del AAAGA	3988	1309
15.12	G→T	4010	1317
15.13	delT	4146	1362
15.13	delGT	4254	1398
15.13	delT	4282	1407
15.14	C→T	4410	1450
15.14	G→T	4452	1464
15.15	delAG	4455	1465
15.19	insCT	5326	1755
15.21	insT	5596	1845
15.22	delG	5940	1960

Table 3 The relationship between the APC mutation and clinicopathological parameters

Parameters	No. of cases	APC mutation
Size
< 5 cm	31	7
> 5 cm	37	8
Differentiation
Well	14	4
Moderate	20	6
Poor	28	3
Mucinous	6	2
Lauren classification
Intestinal type	30	10a
Diffuse type	38	5
Serosal invasion
Absent	30	6
Present	38	9
Metastasis
Absent	33	8
Present	35	7
Clinical stage
I and II	36	9
III and IV	32	6

^aP < 0.05 vs diffuse type

Table 4 The relationship between MSI and APC mutation

MSI status	No. of cases	APC mutation
MSI-H	8	0
MSI-L	9	2
MSS	51	13