利用转录组二代测序探索直肠癌术前放化疗的敏感性分子特征

doi:10.19723/j.issn.1671-167X.2019.03.025

北京大学学报（医学版） ›› 2019, Vol. 51 ›› Issue (3): 542-547. doi: 10.19723/j.issn.1671-167X.2019.03.025

利用转录组二代测序探索直肠癌术前放化疗的敏感性分子特征

张威¹,叶颖江²,任仙文³,黄晶⁴,申占龙^1△()

收稿日期:2019-03-25 出版日期:2019-06-18 发布日期:2019-06-26
作者简介:申占龙,北京大学人民医院胃肠外科主任医师、副教授、博士生导师,外科肿瘤研究室副主任。任中国医师协会外科医师分会多学科综合治疗(multi-disciplinary team, MDT)专业委员会副秘书长、青年委员会主任委员,中国医师协会外科医师分会经肛门全直肠系膜切除术专业委员会(transanal total mesorectal excision, taTME)副主任委员,中国医师协会结直肠肿瘤专业委员会taTME学组副主任委员,中国抗癌协会大肠癌专业委员会青年委员会副主任委员,中华医学会外科学分会实验外科学组委员、中国医师协会外科医师分会亚太胃肠外科研究协作组(Asian-Pacific Study Group for Gastrointestinal Surgery, APGIS)执行主席等。以第一作者或通信作者发表论文60余篇,其中SCI论文37篇,总影响因子140.617。2018年,关于结直肠癌T细胞动态变化的单细胞测序研究成果发表在Nature杂志,获得中国生物信息学十大进展称号。作为课题负责人,主持国家自然科学基金2项,北京市自然科学基金1项,北京大学临床医学+X青年专项基金1项,医院发展基金4项,国家专利7项,荣获北京大学医学部优秀科研创新奖、优秀教师,北京大学人民医院学术新星一等奖、优秀教学研究奖,优秀对外交流奖,韩国胃癌学会最佳报告奖等。临床专长:胃肠肿瘤的诊断与外科治疗,尤其是腹腔镜微创手术,包括结肠癌完整结肠系膜切除术、肛提肌外腹会阴联合切除术、直肠癌超低位保肛taTME手术、胃癌根治术。研究方向:胃肠道肿瘤侵袭转移的机制研究,尤其是与监测预后和治疗敏感性相关的分子标志物的筛选和验证研究,包括蛋白质修饰组学(乙酰化)、非编码RNA(microRNA、lncRNA、circRNA)、肿瘤微环境[肿瘤相关巨噬细胞(tumor associated macrophage,TAM)、T细胞、B细胞]等。
基金资助:
北京大学临床医学+X青年专项(PKU2018LCXQ021)-中央高校基本科研业务费

Detection of preoperative chemoradiotherapy sensitivity molecular characteristics of rectal cancer by transcriptome second generation sequencing

Wei ZHANG¹,Ying-jiang YE²,Xian-wen REN³,Jing HUANG⁴,Zhan-long SHEN^1△()

Received:2019-03-25 Online:2019-06-18 Published:2019-06-26
Supported by:
Supported by the Fundamental Research Funds for the Central Universities: Peking University Clinical Medicine Plus X-Young Scholars Project (PKU2018LCXQ021)

摘要/Abstract

摘要： 目的探索直肠癌新辅助放化疗的敏感性分子特征。方法前瞻性收集局部进展期中、低位直肠癌30例患者的临床病理资料,包括一般情况、放化疗前影像学资料、放化疗前活组织病理检查资料、肿瘤分化程度等9项指标,分析其与直肠癌放化疗后肿瘤消退分级(tumor regression grading,TRG)的相关性。收集这30例患者新辅助治疗前结肠镜活检冰冻标本,进行转录组二代测序和生物信息学分析,筛选可能驱动直肠癌放化疗敏感性的基因。结果 30例直肠癌患者中,病理完全缓解9例,部分缓解12例,缓解差9例。直肠癌放化疗后病理TRG缓解程度与肿瘤术前MRI的T分期呈负相关(P=0.046),与术前MRI直肠癌壁外血管侵犯(extramural vascular invasion, EMVI)呈正相关(P=0.003)。转录组二代测序对所获取的P<0.05的217条转录本进行信号通路富集分析,可以发现多条与抗原呈递相关的细胞信号转导通路,其中HSPA1A、HSPA1B和EXOSC2的高表达和术后病理缓解差呈正相关(P<0.05),DNMBP、WASH8P、FAM57A和SGSM2等的高表达和术后病理缓解良好呈正相关(P<0.05)。结论直肠癌术前MRI检测肿瘤EMVI阳性的患者放化疗后病理完全缓解率明显优于EMVI阴性者。HSPA1A、HSPA1B和EXOSC2高表达的患者术后病理缓解差,而DNMBP、WASH8P、FAM57A和SGSM2高表达的患者术后病理缓解良好。基于直肠癌放化疗敏感性分子特征,尝试阻断或增强与直肠癌放化疗敏感性相关的分子通路,可进一步探索能增加直肠癌放化疗敏感性的候选治疗靶点。

关键词: 直肠肿瘤, 新辅助治疗, 转录组测序, 肿瘤消退分级

Abstract: Objective: To detect the preoperative chemoradiotherapy sensitivity molecular characteristics of rectal cancer by transcriptome second generation sequencing.Methods: The clinicopathological data of 30 patients with locally advanced rectal cancer were collected prospectively, including 9 indicators (general conditions, imaging data before radiotherapy and chemotherapy, pathological data of biopsy before radiotherapy and chemotherapy, and tumor differentiation degree, etc.), in order to analyze the correlation between them and tumor regression grading (TRG) after radiotherapy and chemotherapy for rectal cancer. At the same time, frozen specimens of colonoscopy biopsy before neoadjuvant therapy were collected from these 30 patients, and transcriptome second-generation sequencing was performed for bioinformatics analysis to screen out the genes that might drive the radio chemotherapy sensitivity of rectal can-cer. Results: Among the 30 patients with rectal cancer, 9 had complete pathological remission, 12 had partial remission, and 9 had poor remission. The degree of pathological TRG remission after radiotherapy and chemotherapy for rectal cancer was negatively correlated with the preoperative MRI T stage (P=0.046), and positively correlated with preoperative MRI rectal cancer extravascular invasion (EMVI) (P=0.003). Transcriptome second-generation sequencing of the obtained 217 transcripts (P<0.05) for signal pathway enrichment analysis, and multiple cell signal transduction pathways related to antigen presentation could be found. The high expression of HSPA1A, HSPA1B and EXOSC2 was positively correlated with postoperative pathological remission (P<0.05). The high expression of DNMBP, WASH8P, FAM57A, and SGSM2 was positively correlated with postoperative pathological remission (P<0.05).Conclusion: Preoperative NMR detection of extra-tumoral vascular invasion (EMVI-positive) in patients with rectal cancer was significantly better than that of EMVI-negative patients after chemoradiotherapy. Patients with high expressions of HSPA1A, HSPA1B and EXOSC2 had poor postoperative pathological remission, while patients with high expressions of genes, such as DMNMB, WASH8P, FAM57A, and SGSM2 had good postoperative pathological remission. Based on the molecular characte-ristics of rectal cancer radiotherapy and chemotherapy, attempts to block or enhance the molecular pathways associated with chemosensitivity of rectal cancer, are to be made to further explore new candidate therapeutic targets that can increase the sensitivity of radiotherapy and chemotherapy for rectal cancer.

Key words: Rectal neoplasms, Neoadjuvant therapy, Transcriptome sequencing, Tumor regression grading

中图分类号:

R735.3

张威,叶颖江,任仙文,黄晶,申占龙. 利用转录组二代测序探索直肠癌术前放化疗的敏感性分子特征[J]. 北京大学学报（医学版）, 2019, 51(3): 542-547.

Wei ZHANG,Ying-jiang YE,Xian-wen REN,Jing HUANG,Zhan-long SHEN. Detection of preoperative chemoradiotherapy sensitivity molecular characteristics of rectal cancer by transcriptome second generation sequencing[J]. Journal of Peking University(Health Sciences), 2019, 51(3): 542-547.

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参考文献 20

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Clinical pathological data	Number	Complete remission		Partial emission		Non-remission		P value
Clinical pathological data	Number	n	%	n	%	n	%	P value
Total	30	9	30.0	12	40.0	9	30.0
Gender								0.860
Male	23	6	26.1	10	43.5	7	30.4
Female	7	3	33.3	2	64.3	2	22.2
Age/years								0.732
<65	17	4	23.5	7	41.2	6	35.3
≥65	13	5	38.5	5	38.5	3	23.0
BMI								>0.999
<18.5	1	0	0	1	100.0	0	0
18.5-23.9	11	4	36.4	4	36.4	3	27.2
≥24	18	5	27.8	7	38.9	6	33.3
MRI T staging								0.046
cT1	1	1	100.0	0	0	0	0
cT2	2	1	50.0	1	50.0	0	0
cT3	13	5	38.5	6	46.2	2	15.3
cT4	14	1	7.1	5	35.7	8	57.2
MRI N staging								0.926
cN0	16	5	31.3	6	37.5	5	31.3
cN+	11	3	27.2	4	36.4	4	36.4
Undecided	3	1	33.3	2	66.7	0	0
EMVI								0.003
Positive	17	8	47.1	8	47.1	1	5.8
Negative	13	1	7.7	4	30.8	8	61.5
Tumormaximum diameter								0.898
>5 cm	11	3	27.2	4	36.4	4	36.4
≤5 cm	19	6	31.6	8	42.1	5	26.3
Pathological type								0.867
Adenocarcinoma	21	6	28.6	9	42.4	6	28.6
Mucinous adenocarcinoma	8	3	37.5	2	25.0	3	37.5
Signet ring cell carcinoma	1	0	0	1	100	0	0
Degree of differentiation								0.923
Low	6	1	16.7	3	50.0	2	33.3
Middle	11	3	27.2	5	45.5	3	27.2
High	13	5	38.5	4	30.7	4	30.7

利用转录组二代测序探索直肠癌术前放化疗的敏感性分子特征

Detection of preoperative chemoradiotherapy sensitivity molecular characteristics of rectal cancer by transcriptome second generation sequencing

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