基于体细胞突变数据库筛选免疫原性结直肠癌高频新抗原的方法

doi:10.16288/j.yczz.20-032

遗传 ›› 2020, Vol. 42 ›› Issue (6): 599-612.doi: 10.16288/j.yczz.20-032

• 技术与方法 • 上一篇

基于体细胞突变数据库筛选免疫原性结直肠癌高频新抗原的方法

秦丽丽^1,², 李毅坚¹, 梁兆端¹, 陈蕾¹, 李文慧¹, 陈超^1,^3,⁴, 黄亚灵¹, 张乐^1,^3,⁴, 刘松明^1,^3,⁴, 邱思^1,⁴, 葛玉萍¹, 彭文婷^1,³, 林欣欣^1,⁴, 张秀清^1,⁴, 董旋¹(), 李波^1,⁴()

^1. 深圳华大生命科学研究院，深圳 518083
^2. 大理大学基础医学院，大理 671000
^3. 中国科学院大学华大教育中心，深圳 518083
^4. 武汉华大吉诺因生物科技有限公司，武汉 430079

收稿日期:2020-04-10 修回日期:2020-05-15 出版日期:2020-06-20 发布日期:2020-05-19
通讯作者: 董旋,李波 E-mail:dongxuan@genomics.cn;libo@genomics.cn
作者简介:秦丽丽, 在读硕士研究生,专业方向：病理学和病理生理学。E-mail: veritasdoct168@163.com|李毅坚,硕士,助理研究员,研究方向：肿瘤免疫治疗。E-mail:liyijian@genomics.cn; 秦丽丽和李毅坚为并列第一作者。
基金资助:
国家自然科学基金项目编号(81702826);深圳市科技创新委员会项目资助编号(JCYJ20170303151334808);深圳市科技创新委员会项目资助编号(JCYJ20170817150015170)

A method of screening highly common neoantigens with immunogenicity in colorectal cancer based on public somatic mutation library

Qin Lili^1,², Li Yijian¹, Liang Zhaorui¹, Dai Lei¹, Li Wenhui¹, Chen Chao^1,^3,⁴, Huang Yaling¹, Zhang Le^1,^3,⁴, Liu Songming^1,^3,⁴, Qiu Si^1,⁴, Ge Yuping¹, Peng Wenting^1,³, Lin Xinxin^1,⁴, Zhang Xiuqing^1,⁴, Dong Xuan¹(), Li Bo^1,⁴()

^1. BGI-Shenzhen, Shenzhen 518083, China
^2. College of Basic Medicine, Dali University, Dali 671000, China
^3. BGI Education Center, University of Chinese Academy of Sciences, Shenzhen 518083, China
^4. BGI-GenoImmune, BGI-Shenzhen, Wuhan 430079, China

Received:2020-04-10 Revised:2020-05-15 Online:2020-06-20 Published:2020-05-19
Contact: Xuan Dong,Bo Li E-mail:dongxuan@genomics.cn;libo@genomics.cn
Supported by:
Supported by the National Natural Science Foundation of China No(81702826);Science, Technology and Innovation Commission of Shenzhen Municipality Nos(JCYJ20170303151334808);Science, Technology and Innovation Commission of Shenzhen Municipality Nos(JCYJ20170817150015170)

摘要/Abstract

摘要：

结直肠癌是世界高发和高致死率的恶性肿瘤。靶向新抗原的免疫治疗已被证实可以诱导癌症患者肿瘤持续消退,但这些特异性新抗原,仅适用于个体精准治疗。随着大量的高频肿瘤基因突变被发现,这些与突变相关的高频新抗原可覆盖更多人群,具有较强的临床意义。然而目前结直肠癌中是否也存在高频新抗原仍不清楚。本研究利用来源于321个结直肠癌患者的体细胞突变数据库,联合1种标准过滤和7种预测算法,筛选并获得了25个基于中国人高频分型HLA-A*1101限制性的高频新抗原,它们均具有高亲和力(IC₅₀<50 nmol/L)和高呈递分值(>0.90);其中,除了阳性对照多肽KRAS_G12V_8-16外,11个高频新抗原能够在体外诱导细胞毒性T淋巴细胞(cytotoxic T lymphocyte, CTL)分泌γ干扰素(interferon gamma, IFN-γ),证实具有免疫原性。选取免疫原性最强的新抗原C1orf170_S418G_413-421及阳性对照多肽KRAS_G12V_8-16体外刺激T细胞,利用流式细胞分选及单细胞转录组测序技术,获得其特异性CTL的免疫组库信息,所构建的TCR-T（T-cell receptor engineered T cell）能够识别新抗原并分泌细胞因子。以上结果表明,本研究开发了一种利用体细胞数据库预测并体外筛选验证具有免疫原性高频新抗原的方法,为结直肠癌及其他癌种的多肽、DC（dendritic cells）疫苗、TCR-like抗体、TCR-T等免疫治疗提供了重要的多肽靶点和TCR信息,具有实际的临床应用价值。

关键词: 结直肠癌, 高频, 新抗原, 呈递

Abstract:

Colorectal cancer (CRC) is a malignant cancer with high incidence and mortality in the world. Immunotherapy targeting neoantigens can induce durable tumor regression in cancer patients, but is almost limited to personalized precision therapy, due to the individual differences of unique neoantigens. With the discovery of many common oncogenic mutations, and such mutation-associated neoantigens could cover more patients, and hence are valuable in clinical field. However, whether the common neoantigens can be identified in CRC is unknown. Combining the somatic mutations data from 321 CRC patients with a filter standard and 7 predicted algorithms, we screened and obtained 25 HLA-A*1101-restricted common neoantigens with a high binding affinity (IC₅₀<50 nmol/L) and presentation score (>0.90). Besides the positive epitope KRAS_G12V_8-16, 11 out of 25 common neoantigens specifically induced in vitro pre- stimulated cytotoxic lymphocyte (CTL) to secrete interferon gamma (IFN-γ). Moreover, combining cell-sorting technology and single-cell RNA sequencing, the immune repertoire profiles of C1orf170_S418G_413-421 and KRAS_G12V_8-16-specific CTL were analyzed and validated. Their related T-cell receptor engineered T cell (TCR-T) cells could also recognize the neoantigens and secrete IFN-γ. Hence, we have established a method to screen for common neoantigens with immunogenicity in CRC based on the public somatic mutation library. It can provide essential peptide and TCR information for immunotherapies, such as peptides, dendritic cells (DC) vaccines, TCR-like antibodies, TCR-T, etc., for the CRC and other cancers, which has practical application value in the clinics.

Key words: colorectal cancer, common, neoantigen, presentation

秦丽丽, 李毅坚, 梁兆端, 陈蕾, 李文慧, 陈超, 黄亚灵, 张乐, 刘松明, 邱思, 葛玉萍, 彭文婷, 林欣欣, 张秀清, 董旋, 李波. 基于体细胞突变数据库筛选免疫原性结直肠癌高频新抗原的方法[J]. 遗传, 2020, 42(6): 599-612.

Qin Lili, Li Yijian, Liang Zhaorui, Dai Lei, Li Wenhui, Chen Chao, Huang Yaling, Zhang Le, Liu Songming, Qiu Si, Ge Yuping, Peng Wenting, Lin Xinxin, Zhang Xiuqing, Dong Xuan, Li Bo. A method of screening highly common neoantigens with immunogenicity in colorectal cancer based on public somatic mutation library[J]. Hereditas(Beijing), 2020, 42(6): 599-612.

图/表 7

表1

图1

图2

图3

表2

图4

图5

参考文献 32

[1]	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A . Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2018,68(6):394-424. doi: 10.3322/caac.21492 pmid: 30207593
[2]	Magee MS, Kraft CL, Abraham TS, Baybutt TR, Marszalowicz GP, Li P, Waldman SA, Snook AE . GUCY2C-directed CAR-T cells oppose colorectal cancer metastases without autoimmunity. Oncoimmunology, 2016,5(10):e1227897. doi: 10.1080/2162402X.2016.1227897 pmid: 27853651
[3]	Ciardiello D, Vitiello PP, Cardone C, Martini G, Troiani T, Martinelli E, Ciardiello F . Immunotherapy of colorectal cancer: Challenges for therapeutic efficacy. Cancer Treat Rev, 2019,76:22-32. doi: 10.1016/j.ctrv.2019.04.003 pmid: 31079031
[4]	Parkhurst MR, Yang JC, Langan RC, Dudley ME, Nathan DA, Feldman SA, Davis JL, Morgan RA, Merino MJ, Sherry RM, Hughes MS, Kammula US, Phan GQ, Lim RM, Wank SA, Restifo NP, Robbins PF, Laurencot CM, Rosenberg SA . T cells targeting carcinoembryonic antigen can mediate regression of metastatic colorectal cancer but induce severe transient colitis. Mol Ther, 2011,19(3):620-626. doi: 10.1038/mt.2010.272
[5]	Zhang CC, Wang Z, Yang Z, Wang ML, Li SQ, Li YY, Zhang R, Xiong ZX, Wei ZH, Shen JJ, Luo YL, Zhang QZ, Liu LM, Qin H, Liu W, Wu F, Chen W, Pan F, Zhang XQ, Bie P, Liang HJ, Pecher G, Qian C . Phase I escalating- dose trial of CAR-T therapy targeting CEA + metastatic colorectal cancers . Mol Ther, 2017,25(5):1248-1258. doi: 10.1016/j.ymthe.2017.03.010 pmid: 28366766
[6]	Morgan RA, Yang JC, Kitano M, Dudley ME, Laurencot CM, Rosenberg SA . Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol Ther, 2010,18(4):843-851. doi: 10.1038/mt.2010.24 pmid: 20179677
[7]	Zhou Z, Lyu XZ, Wu JC, Yang XY, Wu SS, Zhou J, Gu X, Su ZX, Chen SQ . TSNAD: an integrated software for cancer somatic mutation and tumour-specific neoantigen detection. R Soc Open Sci, 2017,4(4):170050. doi: 10.1098/rsos.170050 pmid: 28484631
[8]	Nonomura C, Otsuka M, Kondou R, Iizuka A, Miyata H, Ashizawa T, Sakura N, Yoshikawa S, Kiyohara Y, Ohshima K, Urakami K, Nagashima T, Ohnami S, Kusuhara M, Mitsuya K, Hayashi N, Nakasu Y, Mochizuki T, Yamaguchi K, Akiyama Y . Identification of a neoantigen epitope in a melanoma patient with good response to anti- PD-1 antibody therapy. Immunol Lett, 2019,208:52-59. doi: 10.1016/j.imlet.2019.02.004 pmid: 30880120
[9]	Tran E, Robbins PF, Lu YC, Prickett TD, Gartner JJ, Jia L, Pasetto A, Zheng Z, Ray S, Groh EM, Kriley IR, Rosenberg SA . T-Cell transfer therapy targeting mutant KRAS in cancer. N Engl J Med, 2016,375(23):2255-2262. doi: 10.1056/NEJMoa1609279 pmid: 27959684
[10]	Wirth TC, Kühnel F . Neoantigen targeting-dawn of a new era in cancer immunotherapy? Front Immunol, 2017,8:1848. doi: 10.3389/fimmu.2017.01848 pmid: 29312332
[11]	Cafri G, Yossef R, Pasetto A, Deniger DC, Lu YC, Parkhurst M, Gartner JJ, Jia L, Ray S, Ngo LT, Jafferji M, Sachs A, Prickett T, Robbins PF, Rosenberg SA . Memory T cells targeting oncogenic mutations detected in peripheral blood of epithelial cancer patients. Nat Commun, 2019,10(1):449. doi: 10.1038/s41467-019-08304-z pmid: 30683863
[12]	Nielsen M, Lundegaard C, Worning P, Lauemøller SL, Lamberth K, Buus S, Brunak S, Lund O . Reliable prediction of T-cell epitopes using neural networks with novel sequence representations. Protein Sci, 2003,12(5):1007-1017. doi: 10.1110/ps.0239403 pmid: 12717023
[13]	Nielsen M, Andreatta M . NetMHCpan-3.0; improved prediction of binding to MHC class I molecules integrating information from multiple receptor and peptide length datasets. Genome Med, 2016,8(1):33. doi: 10.1186/s13073-016-0288-x pmid: 27029192
[14]	Jurtz V, Paul S, Andreatta M, Marcatili P, Peters B, Nielsen M . NetMHCpan-4.0: improved Peptide-MHC class I interaction predictions integrating eluted ligand and peptide binding affinity data. J Immunol, 2017,199(9):3360-3368. doi: 10.4049/jimmunol.1700893 pmid: 28978689
[15]	Liu G, Li DL, Li Z, Qiu S, Li WH, Chao CC, Yang NB, Li HD, Cheng Z, Song X, Cheng L, Zhang XQ, Wang J, Yang HM, Ma K, Hou Y, Li B . PSSMHCpan: a novel PSSM-based software for predicting class I peptide-HLA binding affinity. Gigascience, 2017,6(5):1-11. doi: 10.1093/gigascience/gix004 pmid: 28327916
[16]	Zhang H, Lund O, Nielsen M . The PickPocket method for predicting binding specificities for receptors based on receptor pocket similarities: application to MHC-peptide binding. Bioinformatics, 2009,25(10):1293-1299. doi: 10.1093/bioinformatics/btp137 pmid: 19297351
[17]	Peters B, Sette A . Generating quantitative models describing the sequence specificity of biological processes with the stabilized matrix method. BMC Bioinformatics, 2005,6:132. doi: 10.1186/1471-2105-6-132 pmid: 15927070
[18]	Creaney J, Ma S, Sneddon SA, Tourigny MR, Dick IM, Leon JS, Khong A, Fisher SA, Lake RA, Lesterhuis WJ, Nowak AK, Leary S, Watson MW, Robinson BW . Strong spontaneous tumor neoantigen responses induced by a natural human carcinogen. Oncoimmunology, 2015,4(7):e1011492. doi: 10.1080/2162402X.2015.1011492 pmid: 26140232
[19]	Hu WP, Li YP, Zhang XQ . MHC-I epitope presentation prediction based on transfer learning.Hereditas(Beijing), 41(11):1041-1049.
	胡伟澎, 李佑平, 张秀清 . 基于迁移学习的MHC-I型抗原表位呈递预测. 遗传, 41(11):1041-1049.
[20]	Hu WP, Qiu S, Li YP, Lin XX, Zhang L, Xiang HT, Han X, Chen L, Li S, Li WH, Ren Z, Hou GX, Lin ZL, Lu JL, Liu G, Li B, Lee LJ . EPIP: MHC-I epitope prediction integrating mass spectrometry derived motifs and tissue- specific expression profile. bioRxiv, 2019,567081.
[21]	Tanyi JL, Bobisse S, Ophir E, Tuyaerts S, Roberti A, Genolet R, Baumgartner P, Stevenson BJ, Iseli C, Dangaj D, Czerniecki B, Semilietof A, Racle J, Michel A, Xenarios I, Chiang C, Monos DS, Torigian DA, Nisenbaum HL, Michielin O, June CH, Levine BL, Powell DJ Jr, Gfeller D, Mick R, Dafni U, Zoete V, Harari A, Coukos G, Kandalaft LE. Personalized cancer vaccine effectively mobilizes antitumor T cell immunity in ovarian cancer. Sci Transl Med, 2018, 10(436): eaao5931. doi: 10.1126/scitranslmed.aao3003 pmid: 29643228
[22]	Yamamiya D, Mizukoshi E, Kaji K, Terashima T, Kitahara M, Yamashita T, Arai K, Fushimi K, Honda M, Kaneko S . Immune responses of human T lymphocytes to novel hepatitis B virus-derived peptides. PLoS One, 2018,13(6):e0198264. doi: 10.1371/journal.pone.0198264 pmid: 29856876
[23]	Rodenko B, Toebes M, Hadrup SR, van Esch WJ, Molenaar AM, Schumacher TN, Ovaa H,. Generation of peptide-MHC class I complexes through UV-mediated ligand exchange. Nat Protoc, 2006,1(3):1120-1132. doi: 10.1038/nprot.2006.121 pmid: 17406393
[24]	Kim MS, Ma JS, Yun HY, Cao Y, Kim JY, Chi V, Wang D, Woods A, Sherwood L, Caballero D, Gonzalez J, Schultz PG, Young TS, Kim CH . Redirection of genetically engineered CAR-T cells using bifunctional small molecules. J Am Chem Soc, 2015,137(8):2832-2835. doi: 10.1021/jacs.5b00106 pmid: 25692571
[25]	Ali M, Foldvari Z, Giannakopoulou E, Böschen ML, Strønen E, Yang W, Toebes M, Schubert B, Kohlbacher O, Schumacher TN, Olweus J . Induction of neoantigen- reactive T cells from healthy donors. Nat Protoc, 2019,14(6):1926-1943. doi: 10.1038/s41596-019-0170-6 pmid: 31101906
[26]	Lancaster EM, Jablons D, Kratz JR . Applications of next-generation sequencing in eoantigen prediction and cancer vaccine development. Genet Test Mol Biomarkers, 2019,24(2):59-66. doi: 10.1089/gtmb.2018.0211 pmid: 30907630
[27]	The problem with neoantigen prediction. Nat Biotechnol, 2017,35(2):97. doi: 10.1038/nbt.3800 pmid: 28178261
[28]	Cohen CJ, Gartner JJ, Horovitz-Fried M, Shamalov K, Trebska-McGowan K, Bliskovsky VV, Parkhurst MR, Ankri C, Prickett TD, Crystal JS, Li YF, El-Gamil M, Rosenberg SA, Robbins PF, . Isolation of neoantigen- specific T cells from tumor and peripheral lymphocytes. J Clin Invest, 2015,125(10):3981-3991. doi: 10.1172/JCI82416 pmid: 26389673
[29]	Lin QY, Liu Z, Luo MJ, Zheng H, Qiao S, Han CL, Deng DQ, Fan Z, Lu YF, Zhang ZH, Luo QM . Visualizing DC morphology and T cell motility to characterize DC-T cell encounters in mouse lymph nodes under mTOR inhibition. Sci China Life Sci, 2019,62(9):1168-1177. doi: 10.1007/s11427-018-9470-9 pmid: 31016533
[30]	Simoni Y, Becht E, Fehlings M, Loh CY, Koo SL, Teng KWW, Yeong JPS, Nahar R, Zhang T, Kared H, Duan K, Ang N, Poidinger M, Lee YY, Larbi A, Khng AJ, Tan E, Fu C, Mathew R, Teo M, Lim WT, Toh CK, Ong BH, Koh T, Hillmer AM, Takano A, Lim TKH, Tan EH, Zhai W, Tan DSW, Tan IB, Newell EW . Bystander CD8 + T cells are abundant and phenotypically distinct in human tumour infiltrates . Nature, 2018,557(7706):575-579. doi: 10.1038/s41586-018-0130-2 pmid: 29769722
[31]	Whiteside SK, Snook JP, Williams MA, Weis JJ . Bystander T Cells: a balancing act of friends and foes. Trends Immunol, 2018,39(12):1021-1035. doi: 10.1016/j.it.2018.10.003 pmid: 30413351
[32]	Kim TS, Shin EC . The activation of bystander CD8 + T cells and their roles in viral infection . Exp Mol Med, 2019,51(12):1-9. doi: 10.1038/s12276-019-0351-y pmid: 31811117

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

预测的新抗原表位	基因	突变位点	突变频率	新抗原突变位置	亲和力 (nmol/L)	EPIP分数	肿瘤中的作用
GLCE_V533I_526-535	GLCE	V533I	6/321	STIDESPIFK	2.7	0.989077	?
C1orf170_S418G_413-421	C1orf170	S418G	5/321	SVAGPGPNK	21.7	0.974471	?
MUC3A_I29T_28-36	MUC3A	I29T	6/321	STSQVPFPR	13.8	0.970609	?
CCRL2_F179Y_174-183	CCRL2	F179Y	5/321	ATLPEYVVYK	4.7	0.968889	?-
KIAA1683_M313T_311-319	KIAA1683	M313T	6/321	STTTTTPPK	7.9	0.964324	?
KLHL40_N345S_341-349	KLHL40	N345S	5/321	ASLSSQVPK	9.4	0.956861	?
MUC3A_S175P_172-181	MUC3A	S175P	6/321	STYPMTTTEK	5.8	0.952998	?
RNF43_I47V_46-54	RNF43	I47V	7/321	AVIRVIPLK	7.8	0.949232	TSG
SYNE2_A2395T_2389-2398	SYNE2	A2395T	5/321	STQESATVEK	28.9	0.946256	?
TLR10_I369L_367-375	TLR10	I369L	5/321	RTLQLPHLK	13.2	0.945037	?
ANKRD36C_N1571S_1571-1579	ANKRD36C	N1571S	5/321	STMEKCIEK	6.6	0.94484	?
IYD_F231I_229-237	IYD	F231I	6/321	QVIGKIILK	21.4	0.940062	?
SSX5_E19Q_12-20	SSX5	E19Q	5/321	RVGSQIPQK	35.7	0.936871	?
MUC3A_S326T_319-327	MUC3A	S326T	7/321	TTLPTTITR	16.1	0.936033	?
ARHGEF11_H1427R_1427-1435	ARHGEF11	H1427R	5/321	RTIEQLTLK	10.8	0.933929	?
CTNNB1_T41A_41-49	CTNNB1	T41A	6/321	ATAPSLSGK	6.9	0.932574	癌基因
LILRB5_L605F_603-611	LILRB5	L605F	5/321	RSFPLTLPR	6.9	0.931333	?
EIF2A_T92S_86-95	EIF2A	T92S	5/321	ATWQPYSTSK	12.9	0.931289	?
TMPRSS15_P732S_732-740	TMPRSS15	P732S	5/321	STDGGPFVK	12	0.930946	?
MUC6_P2049L_2044-2053	MUC6	P2049L	9/321	GTVPPLTTLK	16.5	0.917963	?
TMEM185B_A42G_42-51	TMEM185B	A42G	5/321	GVFAPIWLWK	5.7	0.904248	?
UNC93A_M403T_402-410	UNC93A	M403T	6/321	STFLCVHVK	4.5	0.903415	?
MUC3A_Q31H_28-36	MUC3A	Q31H	5/321	SISHVPFPR	14.1	0.901598	?
FSIP2_R1288Q_1285-1293	FSIP2	R1288Q	5/321	SSLQSQLSK	13.8	0.900572	?
FSIP2_T184NX	FSIP2	T184NX	2/321	TTLPKFNKK	10.2	0.961067	?

抗原肽	TCR克隆	CDR3 序列	TRV 基因	J 基因	比例(%)
C1orf170_S418G_413-421	Clonotype1	TRA: CAASGGAQKLVF	TRAV29DV5	TRAJ54	53.2
		TRA: CAGLLYNSGNTPLVF	TRAV35	TRAJ29
		TRB: CASSRDRGSNQPQHF	TRBV27	TRBJ1-5
	Clonotype2	TRA: CAASGGAQKLVF	TRAV29DV5	TRAJ54	31.6
	Clonotype2	TRB: CASSRDRGSNQPQHF	TRBV27	TRBJ1-5	31.6
KRAS_G12V_8-16	Clonotype1	TRA: CASNDYKLSF	TRAV8-3	TRAJ20	80.5
	Clonotype1	TRB: CASSLDGVSYEQYF	TRBV11-2	TRBJ2-7	80.5
	Clonotype2	TRA: ?	?	?	14.3
	Clonotype2	TRB: CASSLDGVSYEQYF	TRBV11-2	TRBJ2-7	14.3

基于体细胞突变数据库筛选免疫原性结直肠癌高频新抗原的方法

A method of screening highly common neoantigens with immunogenicity in colorectal cancer based on public somatic mutation library

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 7

参考文献 32

相关文章 5

编辑推荐

Metrics

[1]	李以格, 张丹丹. 后GWAS时代结直肠癌致病SNP功能机制的研究进展[J]. 遗传, 2021, 43(3): 203-214.
[2]	卢涣滋,王迪侃,王智. HPV阳性口咽癌患者预后与T细胞浸润和新抗原负荷相关性分析[J]. 遗传, 2019, 41(8): 725-735.
[3]	胡伟澎, 李佑平, 张秀清. 基于迁移学习的MHC-I型抗原表位呈递预测[J]. 遗传, 2019, 41(11): 1041-1049.
[4]	李会晨，冯会媛，张锡朋，刘蕊，马东旺，秦海，周毅，俞林. 天津地区人群hMLH1和hMSH2基因多态性与散发性结直肠癌易感性的关系[J]. 遗传, 2010, 32(12): 1241-1246.
[5]	朱益民，林洁，陈俭，黄琼，邵丽娜，来茂德. Staufen基因在结直肠癌组织中的表达[J]. 遗传, 2005, 27(5): 705-709.