KRAS、NRAS、BRAF及PIK3CA基因突变与结直肠癌患者临床病理特征的关系

doi:10.3877/cma.j.issn.2095-5820.2021.04.005

目的

检测结直肠癌组织KRAS、NRAS、BRAF及PIK3CA基因突变状态，并分析突变与临床病理特征的关系。

方法

收集结直肠癌手术切除或穿刺活检标本189例，提取DNA后，采用突变扩增阻滞系统技术(ARMS)检测KRAS、NRAS、BRAF及PIK3CA基因的突变状态，并分析基因突变与临床特征的关系。

结果

KRAS基因突变率为49.7%，2号外显子突变最常见；NRAS基因突变率为3.7%；BRAF基因突变率为4.2%；PIK3CA基因突变率为2.6%；PIK3CA基因突变者均与KRAS存在共同突变；近端结肠癌患者KRAS突变率显著高于远端结肠癌患者(P=0.048)；无神经束侵犯结肠癌患者NRAS突变率显著高于有神经束侵犯患者(P=0.001)；肿瘤浸润深度T4患者BRAF基因突变率显著高于浸润深度为T1~T3的患者(P=0.029)；低分化肿瘤患者BRAF基因突变率显著高于中高分化肿瘤患者(P=0.023)。

结论

结直肠癌患者KRAS基因突变率较高，KRAS、NRAS和BRAF突变率与结直肠癌临床病理特征存在关联。

关键词: 结直肠肿瘤, 基因, 突变

Objective

To study the relationship between KRAS, NRAS, BRAF and PIK3CA mutations and the clinicopathological features in patients with colorectal cancer (CRC).

Methods

The 189 samples of colorectal cancer were collected by surgical excision or biopsy,and DNA were extracted. The mutations of KRAS, NRAS, BRAF and PIK3CA genes were detected by amplification refractory mutation system, and its relationship with clinicopathological features of CRC was analyzed.

Results

The mutations rate of KRAS were 49.7%, in most exon2 mutations; the mutations rate of NRAS were 3.7%; the mutations rate of BRAF were 4.2%; the mutations rate of PIK3CA were 2.6%. There were 5 patients with PIK3CA gene mutations, all of them had KRAS mutations. KRAS mutations were significantly more prevalent in tumors in the proximal colon than in tumors in the distal colon (P=0.048). NRAS mutations rate in patients without nerve bundle invasion were remarkably higher than in patients with nerve bundle invasion (P=0.001). BRAF mutations were significantly more prevalent in tumors in T4 than in tumors in T1-T3 (P=0.029). BRAF mutations were also more common in poorly differentiated tumors than in well and moderately differentiated tumors (P=0.023).

Conclusions

The mutations rate of KRAS gene is high, KRAS, NRAS and BRAF mutations were significantly correlated with the clinicopathological features of CRC.

Key words: Colorectal cancer, Gene, Mutation

表1 KRAS、NRAS、BRAF及PIK3CA基因突变情况

基因名称	检测区段	突变例数(例)	突变率(%)	合计
	2号外显子	79	41.8	49.7% (94/189)
KARS	3号外显子	6	3.1
	4号外显子	9	4.8

	2号外显子	4	2.1	3.7% (7/189)
NRAS	3号外显子	3	1.6
	4号外显子	0	0.0

BRAF	15号外显子	8	4.2	4.2% (8/189)

PIK3CA	20号外显子	5	2.6	2.6% (5/189)

表1 KRAS、NRAS、BRAF及PIK3CA基因突变情况

基因名称	检测区段	突变例数(例)	突变率(%)	合计
	2号外显子	79	41.8	49.7% (94/189)
KARS	3号外显子	6	3.1
	4号外显子	9	4.8

	2号外显子	4	2.1	3.7% (7/189)
NRAS	3号外显子	3	1.6
	4号外显子	0	0.0

BRAF	15号外显子	8	4.2	4.2% (8/189)

PIK3CA	20号外显子	5	2.6	2.6% (5/189)

表2 KRAS基因突变类型和频率

检测区段	突变类型	例数(例)	合并PIK3CA(例)
2号外显子	G12S，G12D	40	3
2号外显子	G12C，G12R，G12V，G12A，G13C	27	1
2号外显子	G13D	12	1
3号外显子	Q61L，Q61R，Q61H	9	0
4号外显子	K117N，A146T，A146V，A146P	6	0

表2 KRAS基因突变类型和频率

检测区段	突变类型	例数(例)	合并PIK3CA(例)
2号外显子	G12S，G12D	40	3
2号外显子	G12C，G12R，G12V，G12A，G13C	27	1
2号外显子	G13D	12	1
3号外显子	Q61L，Q61R，Q61H	9	0
4号外显子	K117N，A146T，A146V，A146P	6	0

表3 KRAS、NRAS、BRAF和PIK3CA基因在不同临床病理特征中的突变情况(例)

项目	总例数 n=189	KRAS			NRAS			BRAF			PIK3CA
项目	总例数 n=189	突变	野生	P值	突变	野生	P值	突变	野生	P值	突变	野生	P值
年龄				0.948			1.000			0.968			0.510
＞60岁	105	52	53		4	101		5	100		4	101
≤60岁	84	42	42		3	81		3	81		1	83
性别				0.491			1.000			0.761			0.625
男	116	60	56		4	112		4	112		2	114
女	73	34	39		3	70		4	69		3	70
神经束侵犯				0.714			0.001			0.820			0.585
有	30	14	16		0	30		2	28		1	29
无	159	80	79		7	152		6	153		4	155
脉管癌栓				0.208			0.949			0.871			0.617
有	52	22	30		2	50		2	50		2	50
无	137	72	65		5	132		6	131		3	134
肿瘤大小				0.406			0.783			0.064			0.973
≥5 cm	76	35	41		2	74		6	70		2	74
＜5 cm	113	59	54		5	108		2	111		3	110
临床分期				0.625			0.618			0.851			0.525
Ⅰ期	26	16	10		0	26		0	26		0	26
Ⅱ期	57	27	30		2	55		3	54		2	55
Ⅲ期	61	30	31		4	57		3	58		3	58
Ⅳ期	45	21	24		1	44		2	43		0	45
肿瘤浸润深度				0.505			0.598			0.029			0.849
T1~2	32	18	14		0	32		0	32		0	32
T3	98	50	48		4	94		2	96		3	95
T4	59	26	33		3	56		6	53		2	57
淋巴结转移				0.894			0.417			0.515			0.062
N0	89	45	44		2	87		4	85		1	88
N1	55	28	27		2	53		1	54		4	51
N2	45	21	24		3	42		3	42		0	45
分化程度				0.088			0.549			0.023			1.000
高分化	6	3	3		0	6		0	6		0	6
中分化	169	88	81		6	163		5	164		5	164
低分化	14	3	11		1	13		3	11		0	14
根治程度				0.890			0.350			0.836			0.592
根治	148	74	74		7	141		7	141		5	143
姑息	41	20	21		0	41		1	40		0	41
肿瘤部位				0.048			0.254			0.085			0.166
近端结肠	46	28	18		0	46		3	43		3	43
远端结肠	82	35	47		3	79		5	77		1	81
直肠	61	31	30		4	57		0	61		1	60

表3 KRAS、NRAS、BRAF和PIK3CA基因在不同临床病理特征中的突变情况(例)

项目	总例数 n=189	KRAS			NRAS			BRAF			PIK3CA
项目	总例数 n=189	突变	野生	P值	突变	野生	P值	突变	野生	P值	突变	野生	P值
年龄				0.948			1.000			0.968			0.510
＞60岁	105	52	53		4	101		5	100		4	101
≤60岁	84	42	42		3	81		3	81		1	83
性别				0.491			1.000			0.761			0.625
男	116	60	56		4	112		4	112		2	114
女	73	34	39		3	70		4	69		3	70
神经束侵犯				0.714			0.001			0.820			0.585
有	30	14	16		0	30		2	28		1	29
无	159	80	79		7	152		6	153		4	155
脉管癌栓				0.208			0.949			0.871			0.617
有	52	22	30		2	50		2	50		2	50
无	137	72	65		5	132		6	131		3	134
肿瘤大小				0.406			0.783			0.064			0.973
≥5 cm	76	35	41		2	74		6	70		2	74
＜5 cm	113	59	54		5	108		2	111		3	110
临床分期				0.625			0.618			0.851			0.525
Ⅰ期	26	16	10		0	26		0	26		0	26
Ⅱ期	57	27	30		2	55		3	54		2	55
Ⅲ期	61	30	31		4	57		3	58		3	58
Ⅳ期	45	21	24		1	44		2	43		0	45
肿瘤浸润深度				0.505			0.598			0.029			0.849
T1~2	32	18	14		0	32		0	32		0	32
T3	98	50	48		4	94		2	96		3	95
T4	59	26	33		3	56		6	53		2	57
淋巴结转移				0.894			0.417			0.515			0.062
N0	89	45	44		2	87		4	85		1	88
N1	55	28	27		2	53		1	54		4	51
N2	45	21	24		3	42		3	42		0	45
分化程度				0.088			0.549			0.023			1.000
高分化	6	3	3		0	6		0	6		0	6
中分化	169	88	81		6	163		5	164		5	164
低分化	14	3	11		1	13		3	11		0	14
根治程度				0.890			0.350			0.836			0.592
根治	148	74	74		7	141		7	141		5	143
姑息	41	20	21		0	41		1	40		0	41
肿瘤部位				0.048			0.254			0.085			0.166
近端结肠	46	28	18		0	46		3	43		3	43
远端结肠	82	35	47		3	79		5	77		1	81
直肠	61	31	30		4	57		0	61		1	60

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Relationship between KRAS, NRAS, BRAF and PIK3CA gene mutations and clinicopathological features in colorectal cancer

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Relationship between KRAS, NRAS, BRAF and PIK3CA gene mutations and clinicopathological features in colorectal cancer