文章摘要

CRIP1对肝癌诊断的临床意义及其对肝癌细胞增殖和转移的影响

作者: 1黄雯, 2仰丽丽, 3吴付兵
1 南京医科大学第四附属医院肿瘤科,南京 210000
2 南京市第二医院肿瘤科,南京 210000
3 南京医科大学附属逸夫医院肿瘤科,南京 210000
通讯: 吴付兵 Email: wfbwkx@163.com
DOI: 10.3978/j.issn.2095-6959.2020

摘要

目的:探讨富含半胱氨酸的肠道蛋白1(cysteine-rich intestinal protein 1,CRIP1)对肝癌诊断的临床意义及其对肝癌细胞增殖和转移的影响。方法:生物信息学在线工具“Gene Expression Profiling Interactive Analysis (GEPIA)”分析CRIP1在肝癌组织和癌旁正常组织中的表达。qRT-PCR检测血清样本中CRIP1 mRNA相对表达水平。qRT-PCR和蛋白质印迹法分别检测转染CRIP1的小干扰RNA(small interfering RNA,siRNA)序列后肝癌细胞中CRIP1 mRNA和蛋白表达。MTT实验和Transwell实验分别检测干扰CRIP1对肝癌细胞增殖和转移的影响。结果:“GEPIA”分析结果表明CRIP1在肝癌组织中的相对表达水平显著高于在癌旁正常组织。与健康体检者相比,肝癌患者血清中CRIP1 mRNA相对表达水平明显升高。以血清中CRIP1 mRNA表达水平诊断肝癌的受试者工作特征(receiver operating characteristic,ROC)曲线下面积为0.901。肝癌患者血清中CRIP1 mRNA相对表达水平与患者性别、年龄无关,但与肿瘤大小、TNM分期显著相关。转染si-CRIP1能有效干扰Hep3B和SMMC7721细胞中CRIP1 mRNA和蛋白的表达。和转染si-Negative Control (si-NC)组相比,转染si-CRIP1能显著抑制Hep3B和SMMC7721细胞的增殖、迁移和侵袭。结论:血清中CRIP1 mRNA表达水平诊断肝癌具有一定的临床意义。干扰CRIP1表达显著抑制肝癌细胞增殖、迁移和侵袭。
关键词: 肝癌;富含半胱氨酸的肠道蛋白1;临床意义;增殖;转移

Clinical significance of CRIP1 in diagnosis of hepatocellular carcinoma and its effect on proliferation and metastasis of hepatocellular carcinoma cells

Authors: 1HUANG Wen, 2YANG Lili, 3WU Fubing
1 Department of Oncology, Fourth Affiliated Hospital of Nanjing Medical University, Nanjing 210000, China
2 Department of Oncology, Nanjing Second Hospital, Nanjing 210000, China
3 Department of Oncology, Yifu Hospital Affiliated to Nanjing Medical University, Nanjing 210000, China

CorrespondingAuthor: WU Fubing Email: wfbwkx@163.com

Abstract

Objective: To investigate the clinical significance of cysteine-rich intestinal protein 1 (CRIP1) in diagnosis of hepatocellular carcinoma and its effects on proliferation and metastasis of hepatocellular carcinoma cells. Methods: The expression of CRIP1 in hepatocellular carcinoma and adjacent normal tissues was analyzed by “Gene Expression Profiling Interactive Analysis (GEPIA)”, an online bioinformatics tool. The relative mRNA expression level of CRIP1 in serum samples was detected by qRT-PCR. The expression mRNA and protein of CRIP1 in hepatocellular carcinoma cells transfected with small interfering RNA (siRNA) interfering CRIP1 sequence were measured by qRT-PCR and Western blot, respectively. The effect of interfering CRIP1 on the proliferation and metastasis of hepatocellular carcinoma cells were assessed by MTT and Transwell assay, respectively. Results: “GEPIA” analysis showed that the relative expression level of CRIP1 in hepatocellular carcinoma tissues was significantly higher than that in normal adjacent tissues. The relative mRNA expression level of CRIP1 in serum of patients with hepatocellular carcinoma was significantly increased compared with that in healthy controls. The area under the receiver operating characteristic (ROC) curve of hepatocellular carcinoma diagnosed by serum CRIP1 mRNA expression was 0.901. The relative mRNA expression levels of CRIP1 in serum of patients with hepatocellular carcinoma was not related to gender and age, but was significantly related to tumor size and TNM stage. The transfection of si-CRIP1 could effectively interfere with the mRNA and protein expression of CRIP1 in Hep3B and SMMC7721 cells. Transfected with si-CRIP1 significantly inhibited cell proliferation, migration, and invasion of Hep3B and SMMC7721 cells compared with that transfected with si-Negative Control (si-NC). Conclusion: The mRNA expression level of CRIP1 in serum has certain clinical significance in the diagnosis of hepatocellular carcinoma. Interfering CRIP1 expression significantly inhibits the proliferation, migration and invasion of hepatocellular carcinoma cells.
Keywords: hepatocellular carcinoma; Cysteine-rich intestinal protein 1; clinical significance; proliferation; metastasis