司仙科 李炜 于昆 张计训 曹亦军 杨佳华

摘 要 目的：构建针对人几丁质酶-3样蛋白-1（chitinase-3-like protein-1， CHI3L1）基因的短发卡RNA（short hairpin RNA， shRNA）干扰表达载体，观察CHI3L1基因表达对人胃癌细胞株BGC-823细胞增殖和侵袭能力的影响。方法：根据人CHI3L1基因序列设计并构建CHI3L1基因的shRNA真核表达载体，通过荧光显微镜观察转染效果。应用逆转录-聚合酶链反应和Western-Blot方法检测BGC-823细胞的CHl3L1 mRNA和CHl3L1基因的表达，通过平板克隆形成实验观察BGC-823细胞的增殖情况，通过Transwell实验观察BGC-823细胞侵袭能力的变化。结果：建立了稳定表达CHI3L1 shRNA的BGC-823细胞的CHI3L1 mRNA和CHI3L1基因的表达均显著减少，且见CHI3L1 shRNA可显著抑制BGC-823细胞的增殖和侵袭能力。结论：干扰CHI3L1基因的表达可显著抑制BGC-823细胞的增殖和侵袭能力。

关键词 胃癌 人几丁质酶-3样蛋白-1 增殖 侵袭

中图分类号：R735.2 文献标志码：A 文章编号：1006-1533（2018）21-0062-04

Expression of human chitinase-3-like protein-1 in gastric cancer and its effect on proliferation and invasion of gastric cancer cells*

SI Xianke**， LI Wei， YU Kun， ZHANG Jixun， CAO Yijun， YANG Jiahua***（Department of General Surgery， Putuo Hospital， Shanghai University of Traditional Chinese Medicine， Shanghai 200062， China）

ABSTRACT Objective： To construct a short hairpin RNA （shRNA） interference expression vector containing a gene encoding human chitinase-3-like protein-1 （CHI3L1） and observe the effect of CHI3L1 gene expression on the proliferation and invasion of human gastric cancer cell line BGC-823. Methods： The shRNA eukaryotic expression vector containing a gene encoding human CHI3L1 was designed and constructed based on CHI3L1 gene sequence and the transfection effect was observed by fluorescence microscopy. CHl3L1 mRNA and its gene expression in BGC-823 cells were detected by reverse transcriptionpolymerase chain reaction and Western-Blot. The proliferation and the invasion ability of BGC-823 cells were observed by clonogenic assay and Transwell method. Results： CHI3L1 mRNA and CHI3L1 expression in BGC-823 cells were significantly reduced and the proliferation and the invasion ability of BGC-823 cells were significantly inhibited by CHI3L1 shRNA. Conclusion： The expression of interference CHI3L1 can significantly inhibit the proliferation and invasion of gastric cancer cell.

KEy WORDS gastric cancer； human chitinase-3-like protein-1； proliferation； invasion

目前認为，胃癌的侵袭、转移是一个连续、复杂的主动过程，与多种相关基因和分子标志物有关[1]。既往研究发现，人几丁质酶-3样蛋白-1（chitinase-3-like protein-1， CHI3L1）在多种肿瘤的增殖、侵袭和转移过程中起着重要作用[2-3]。但对人CHI3L1在人胃癌细胞中的表达及其可能的意义却迄今尚无系统性的研究。本实验通过构建靶向干扰人CHl3L1的重组慢病毒表达载体，观察其对人胃癌细胞增殖和侵袭能力的影响。

1 材料与方法

1.1 材料

1.1.1 细胞

人胃癌细胞株BGC-823细胞和人胚肾上皮细胞株HEK 293T细胞（均由上海交通大学医学院附属新华医院提供）。

1.1.2 主要试剂与仪器

BCA蛋白定量试剂盒、化学发光检测试剂盒和蛋白电泳制胶所需试剂（均为上海秀材生物技术有限公司产品）；……