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中华结直肠疾病电子杂志

中华结直肠疾病电子杂志 ›› 2018, Vol. 07 ›› Issue (02) : 130 -136. doi: 10.3877/cma.j.issn.2095-3224.2018.02.006

所属专题： 文献；

论著

PLAGL2与结肠癌侵袭能力的相关性研究
丁秀杰1, 王永鹏2,†(), 马思平2, 张睿2, 王燕3, 张庆彤2, 张昊2, 李鹏雷2, 林涛2, 李岩溪2, 陈玉泽2, 刘放2   
  1. 1. 110042　沈阳，辽宁省肿瘤医院核医学
    2. 110042　沈阳，中国医科大学肿瘤医院结直肠外科；110042　沈阳，辽宁省肿瘤医院结直肠外科
    3. 110042　沈阳，辽宁省肿瘤医院中心实验室
  • 收稿日期:2017-08-29 出版日期:2018-04-25
  • 通信作者: 王永鹏
  • 基金资助:
    辽宁省自然科学基金(No.201602447)

Pleomorphic adenoma gene like-2 enhances the invasive ability of cancer cells in human colon adenocarcinoma

Xiujie Ding1, Yongpeng Wang2,†(), Siping Ma2, Rui Zhang2, Yan Wang3, Qingtong Zhang2, Hao Zhang2, Penglei Li2, Tao Lin2, Yanxi Li2, Yuze Chen2, Fang Liu2   

  1. 1. Department of Nuclear Medicine, Liaoning Cancer Hospital and Institute
    2. Department of Colorectal Surgery, Cancer Hospital of China Medical University; Department of Colorectal Surgery, Liaoning Cancer Hospital and Institute
    3. Department of Central Laboratory, Liaoning Cancer Hospital and Institute, Shenyang 110042, China
  • Received:2017-08-29 Published:2018-04-25
  • Corresponding author: Yongpeng Wang
  • About author:
    Corresponding author: Wang Yongpeng, Email:
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引用本文：

丁秀杰, 王永鹏, 马思平, 张睿, 王燕, 张庆彤, 张昊, 李鹏雷, 林涛, 李岩溪, 陈玉泽, 刘放. PLAGL2与结肠癌侵袭能力的相关性研究[J]. 中华结直肠疾病电子杂志, 2018, 07(02): 130-136.

Xiujie Ding, Yongpeng Wang, Siping Ma, Rui Zhang, Yan Wang, Qingtong Zhang, Hao Zhang, Penglei Li, Tao Lin, Yanxi Li, Yuze Chen, Fang Liu. Pleomorphic adenoma gene like-2 enhances the invasive ability of cancer cells in human colon adenocarcinoma[J]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2018, 07(02): 130-136.

目的

探讨多形性腺瘤基因样因子2（PLAGL2）在结肠癌和癌旁组织中的表达水平以及与结肠癌临床病理因素之间的关系，研究PLAGL2对结肠癌细胞侵袭能力的影响及其机制。

方法

通过免疫组化检测40例新鲜结肠癌组织及40例癌旁组织中PLAGL2的表达情况，根据术前影像检查结果及术后组织病理结果判断病人分期情况，分析PLAGL2与临床病理因素之间的关系。构建重组质粒pcDNA3.1-PLAGL2，利用转染试剂脂质体2000对SW480细胞进行转染PLAGL2质粒及对照组空载体质粒，分SW480、Vector、PLAGL2三组进行细胞培养，通过Transwell侵袭实验比较三组细胞侵袭能力的差异，并通Western-blot方法检测三组细胞中E-cadherin、vimentin及MMP-7的表达情况。

结果

PLAGL2在Ⅲ~Ⅳ期、Ⅰ~Ⅱ期结肠癌组织与癌旁组织中的免疫组化评分（IS值）分别为8.12±2.997、5.71±2.494、2.17±0.984，差异有统计学意义（F=64.225，P<0.01）。卡方检验结果提示PLAGL2表达增高的病例临床分期较晚（χ2=5.700，P=0.017）、肿瘤较大（χ2=8.174，P=0.008）以及更易发生远隔脏器转移（χ2=13.610，P<0.001）；Transwell侵袭实验比较细胞侵袭个数，PLAGL2组明显高于SW480和Vector两组，差异有统计学意义（F=27.997，P<0.01）；Western-blot方法检测PLAGL2组细胞中E-cadherin表达明显降低（F=38.461，P<0.01）、vimentin（F=28.741，P<0.01）及MMP-7表达升高（F=24.520，P<0.01），差异有统计学意义。

结论

PLAGL2的表达上调与结肠癌的发生、发展密切相关，并且能够促进结肠癌细胞的侵袭能力，可能机制与诱发上皮间质化（EMT）形成有关。

关键词: 结肠肿瘤, 多形性腺瘤基因样因子2, 上皮间质化, 侵袭
Objective

To investigate the expression of PLAGL2 in colon cancer tissues and adjacent normal tissues and its relationship with the clinicopathological factors of colon cancer, to study the function of PLAGL2 on invasive process of colon cancer cells and its mechanism.

Methods

The expressions of PLAGL2 were detected in 40 fresh colon cancer tissues and control normal tissues by immunohistochemistry. According to the results of preoperative imaging examinations and postoperative pathological staging of patients, the relationship between the expression of PLAGL2 and the clinicopathological factors was analyzed. After construction of the recombinant plasmid pcDNA3.1-PLAGL2, SW480 cells were transfected with the plasmid pcDNA3.1- PLAGL2 and empty vector using Lipofectamine? 2000. Three groups of cell line including SW480, Vector and PLAGL2 were cultured and compared the difference of invasive ability by Transwell assay. Finally, Western-blot detected the expression of E-cadherin, vimentin and MMP-7 among cell line of the three groups.

Results

The immunoreactivity scores of PLAGL2 in stage Ⅲ-Ⅳ, stage Ⅰ-Ⅱ and adjacent normal tissues were 8.12±2.997、5.71±2.494、2.17±0.984 respectively, indicating a statistically significant difference (F=64.225, P<0.01). Chi square test showed that the patients with PLAGL2-over expression had more advanced stages (χ2=5.700, P=0.017)、bigger sizes (χ2=8.174, P=0.008) and easier to happen metastasis (χ2=13.610, P<0.001). The invasion of the cells were significantly promoted in PLAGL2 group than other two groups by Transwell assay (F=27.997, P<0.01). Western-blot method for detection of PLAGL2 cells significantly decreased expression of E-cadherin (F=38.461, P<0.01) and increased the expression of vimentin (F=28.741, P<0.01) and MMP-7 (F=24.520, P<0.01), with statistically significant difference compared other two groups.

Conclusions

The upregulation of PLAGL2, which is closely correlated to the occurrence and development of colon cancer and can promote the invasive ability of colon cancer cells, may be related to the formation of EMT.

Key words: Colonic neoplasms, Pleomorphic adenoma gene like-2, Epithelial mesenchymal transition, Invasion
图1 PLAGL2在不同分期结肠癌和癌旁组织中的表达情况（400×）：1A图为癌旁组织、1B~E图为Ⅰ~Ⅳ期结肠癌组织，PLAGL2表达依次升高且表达位置主要在细胞浆中
表1 结肠癌组织和癌旁组织中IS值的比较
分组 例数 IS值（Mean±SD）
癌旁组织 40 2.170±0.984
Ⅰ~Ⅱ期结肠癌组织 14 5.710±2.494
Ⅲ~Ⅳ期结肠癌组织 26 8.120±2.997
整体比较F，P ? 64.225，<0.001
多重比较LSD-t，P 癌旁组织vs.Ⅰ~Ⅱ期 3.564，<0.001
? 癌旁组织vs.Ⅲ~Ⅳ期 5.965，<0.001
? Ⅰ~Ⅱ期vs.Ⅲ~Ⅳ期 2.401，0.020
表2 PLAGL2与临床病理因素的相关性
临床病理因素 例数 PLAGL2表达水平 χ2值 P值
低表达 高表达
年龄（岁） ? ? ? 0.317 0.729
? ≥55 24 8 16 ? ?
? <55 16 4 12 ? ?
肿瘤大小 ? ? ? 8.174 0.008
? ≥3 cm 29 5 24 ? ?
? <3 cm 11 7 4 ? ?
临床分期 ? ? ? 5.700 0.017
? I~II 14 8 6 ? ?
? III~IV 26 4 22 ? ?
组织学类型 ? ? ? ? 0.997*
? 腺癌 25 8 17 ? ?
? 黏液腺癌 11 3 8 ? ?
? 未分化癌 4 1 3 ? ?
图5 Transwell法观察三组细胞的侵袭能力：上图为迁移至微孔膜下层的细胞情况，由左到右依次为SW480、Vector和PLAGL2（200×）；下图为穿透细胞个数比较（**P<0.01）
图6 Western-blot检测三组细胞株的E-cadherin、vimentin和MMP-7的表达情况及光密度比值的对比分析（**P<0.01）
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