PCR-SSCP-DNA sequencing method in detecting PTEN gene mutation and its significance in human gastric cancer

Table 1 Primer sequence, length and annealing temperature of exons 5-8 in PTEN gene

	Primer sequence	Primer length (bp)	Amplification fragment length (bp)	Annealing temperature (°C)
Exon-5F:	ACCTGTTAAGTTTGTATGCAAC	22	379	52
R	TCCAGGAAGAGGAAAGGAAA	20
Exon-6F:	CATAGCAATTTAGTGAAATAACT	23	274	52
R	GATATGGTTAAGAAAACTGTTC	22
Exon-7F:	TGACAGTTTGACAGTTAAAGG	21	263	58
R	GGATATTTCTCCCAATGAAAG	21
Exon-8F:	CTCAGATTGCCTTATAATAGTC	22	558	52
R	TCTGTTACTTGCTACGTAAAC	21

Table 2 PTEN protein expression in gastric cancer and paracancerous tissue samples and intensity distribution n (%)

Clinicopathological parameters	Cases	PTEN protein expression				P
		-	+	++	+++
Paracancerous	53	0 (0.0)	10 (18.9)	18 (34.0)	25 (47.2)	< 0.005
Gastric cancer	53	18 (34.0)	17 (32.1)	15 (28.3)	3 (5.7)
Differentiation extent						< 0.005
Moderate and high differentiation	26	4 (15.4)	6 (23.1)	10 (38.5)	6 (23.1)
Low differentiation	27	14 (51.9)	8 (29.6)	3 (11.1)	2 (7.4)

Table 3 Correlation between PTEN protein expression and clinicopathological parameters in gastric cancer patients

Clinicopathology parameters	Case (n)	PTEN protein expression (n)		Positive rate (%)	P
		Negative	Positive
Tissues
Paracancerous	53	0	53	100.0	< 0.005
Gastric cancer	53	18	35	66.0
Gender
Male	41	15	26	63.4
Female	12	3	9	66.7
Age (yr)
≤ 60	14	5	9	64.3
> 60	39	13	26	66.7
Size (cm)
≤ 3	16	6	10	68.8
> 3	37	12	25	64.9
Location
Antrum	30	10	20	66.7
Gastric body and cardia	23	8	15	65.2
Infiltrating depth
T1, T2	14	1	13	92.9	< 0.025
T3	15	6	9	60.0
T4	24	11	13	54.2
Lymph node metastasis
Without	21	2	19	90.5	< 0.01
With	32	16	16	50.0
Distant metastasis
Without	45	15	30	66.7
With	8	3	5	62.5
Embolization
Without	8	1	7	87.5
With	45	15	30	66.7
Differentiation extent
Moderate and high	26	4	22	84.6	< 0.025
Low	27	14	13	48.1
PTNM staging
I, II	19	1	18	94.7	< 0.005
III, IV	34	17	17	50.0