肝内胆管细胞癌中hsa_circ_0089153的表达及其生物学功能  

Expression of hsa_circ_0089153 in intrahepatic cholangiocarcinoma and its biological function

作　　者：陈小彬 吴嘉艺 黄理铭 白燕南[1,3] 唐忠 严茂林[1,3] CHEN Xiaobin;WU Jiayi;HUANG Liming;BAI Yannan;TANG Zhong;YAN Maolin(Shengli Clinical Medical College of Fujian Medical University,Fuzhou 350001,China;Department of General Surgery,Nanping First Hospital,Nanping,Fujian 353000,China;Department of Hepatobiliary Surgery,Fujian Provincial Hospital,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床医学院,福建福州350001 [2]福建省南平市第一医院普通外科,福建南平353000 [3]福建省立医院肝胆外科,福建福州350001

出　　处：《中国普通外科杂志》2022年第2期199-207,共9页China Journal of General Surgery

基　　金：福建省自然科学基金资助项目(2017J01244);福建省卫生计生委医学创新课题基金资助项目(2018-CX-8)。

摘　　要：背景与目的:肝内胆管细胞癌(ICC)是仅次于肝细胞癌的第二大肝脏恶性肿瘤,其发病隐匿、恶性度高、预后差。环状RNA (circ RNA)异常表达与多种恶性肿瘤的发生发展关系密切,但circ RNA与ICC的关系研究较少。因此,本研究初步探讨ICC中差异表达circ RNA及其生物学功能。方法:采用福建省立医院经病理确诊的5例ICC患者标本,通过高通量circ RNA芯片技术筛选出ICC及对应癌旁组织中差异表达的circ RNA,进一步在长度500~1 500 bp之间选取其中变化最为明显的circ RNA,并通过q RT-PCR进行验证;然后在HCCC-9810、 RBE细胞系中通过慢病毒过表达该circ RNA,应用CCK-8实验、划痕实验、Transwell实验和流式细胞术分析该circ RNA对ICC细胞生物学功能的影响。结果:与癌旁正常组织比较,ICC中共有76个差异表达的circ RNA,包括63个上调与13个下调的circ RNA,其中低表达的hsa_irc_089153变化倍数最大(FC=103.214);q RT-PCR验证结果与基因芯片结果一致。hsa;irc;089153过表达后,ICC细胞的增殖活性明显增强[HCCC-9810:过表达组(114.816±5.138) vs.空载组(100.000±4.897), RBE:过表达组(117.235±7.079) vs.空载组(100.000±14.896)(均P<0.05)];凋亡率明显降低[HCCC-9810:过表达组(11.277±0.243) vs.空载组(16.673±0.660),RBE:过表达组(4.527±0.301) vs.空载组(6.793±0.534)(均P<0.05)];迁移能力与侵袭能力明显增强[HCCC-9810:过表达组(9.070±1.090) vs.空载组(6.206±1.197),RBE:过表达组(6.197±0.687) vs.空载组(4.198±1.170)(均P<0.05)与HCCC-9810:过表达组(0.370±0.026) vs.空载组(0.274±0.014),RBE:过表达组(0.297±0.016) vs.空载组(0.250±0.019)(均P<0.05)]。结论:ICC中存在多种差异表达的circ RNA,其中hsa;irc;089153在ICC组织中呈明显低表达,但与多数情况不同,hsa_irc_089153过表达对ICC的恶性生物学行为有促进作用,因此,hsa_irc_089153在ICC中的生物学功能及其关联的调控网络仍需进一步探讨。Background and Aims:After hepatocellular carcinoma,intrahepatic cholangiocarcinoma(ICC) is the second most common malignant tumor of the liver,with an insidious onset,high malignant level,and a unfavorable prognosis.Although aberrant circ RNA expression is linked to the onset and progression of several malignant tumors,the association between circ RNA and ICC is rarely investigated.As a result,this study was conducted as a preliminary test to examine circ RNA’s differential expression in ICC and its biological function.Methods:By using high-throughput circ RNA chip technology,five specimens of ICC patients confirmed by pathology in Fujian Provincial Hospital were used to screen out differentially expressed circ RNA in ICC and corresponding adjacent tissues,and the circ RNA with the most obvious change was selected from 500-1500 bp in length and verified by q RT-PCR.The circ RNA was then overexpressed in HCCC-9810 and RBE cell lines using lentivirus,and the impact of the circ RNA on the biological function of ICC cells was investigated using CCK-8,scratch,Transwell,and flow cytometry.Results:Compared with normal tissues adjacent to cancer,there are 76 differentially expressed circ RNAs in ICC,including 63 upregulated and 13 downregulated circ RNAs.The downregulated hsa_circ_0089153 had a highest fold-change(FC=103.214).The results of q RT-PCR were consistent with those of gene chip.After hsa_circ_0089153 overexpression,the proliferation activity of ICC cells significantly increased [HCCC-9810:over-expression group(114.816±5.138) vs.no-load group(100.000±4.897),RBE:over-expression group(117.235±7.079) vs.no-load group(100.000±14.896)(all P<0.05)].The apoptosis rate significantly decreased [HCCC-9810:over-expression group(11.277±0.243) vs.no-load group(16.673±0.660)(all P<0.05)].The migrated and invasive ability significantly increased [HCCC-9810:over-expression group(9.070±1.090) vs.no-load group(6.206±1.197),RBE:over-expression group(6.197±0.687) vs.no-load group(4.198±1.170)(all P<0.05) and HCCC-9810:

关 键 词：胆管肿瘤 胆管 肝内 RNA 环状 细胞增殖 肿瘤浸润 

分 类 号：R735.8[医药卫生—肿瘤]