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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 (06) : 531 -537. doi: 10.3877/cma.j.issn.2095-3224.2018.06.006

所属专题： 文献；

论著

sFRP2和Wnt/β-catenin通路在结直肠癌发生发展中的作用
史友权1, 崇杨2, 汤东3, 熊清泉4, 黄玉琴5, 周怀成2, 张琪2, 金芝祥5, 王道荣3,†()   
  1. 1. 221700　徐州，江苏省丰县人民医院普通外科
    2. 225001　扬州，江苏省苏北人民医院胃肠中心；225001　扬州大学医学院
    3. 225001　扬州，江苏省苏北人民医院胃肠中心
    4. 225001　扬州，江苏省苏北人民医院胃肠中心；410013　长沙，中南大学湘雅医学院
    5. 225001　扬州，江苏省苏北人民医院胃肠中心；116044　大连医科大学
  • 收稿日期:2017-08-23 出版日期:2018-12-25
  • 通信作者: 王道荣
  • 基金资助:
    重点病种规范化诊疗项目(No.BE2015664)

The role of sFRP2 and Wnt /β-catenin pathway in the development and progression of colorectal cancer

Youquan Shi1, Yang Chong2, Dong Tang3, Qingquan Xiong4, Yuqin Huang5, Huaicheng Zhou2, Qi Zhang2, Zhixiang Jin5, Daorong Wang3,†()   

  1. 1. Department of General Surgery of Feng Xian People′s Hospital of Jiangsu Province, Xuzhou 221700, China
    2. Department of General Surgery of Subei People′s Hospital of Jiangsu province, Institute of General Surgery, Yangzhou 225001, China;Yangzhou University Medical Academy, Yangzhou 225001, China
    3. Department of General Surgery of Subei People′s Hospital of Jiangsu province, Institute of General Surgery, Yangzhou 225001, China
    4. Department of General Surgery of Subei People′s Hospital of Jiangsu province, Institute of General Surgery, Yangzhou 225001, China;Xianya School of Medicine, Changsha 410013, China
    5. Department of General Surgery of Subei People′s Hospital of Jiangsu province, Institute of General Surgery, Yangzhou 225001, China;Dalian Medical University, Dalian 116044, China
  • Received:2017-08-23 Published:2018-12-25
  • Corresponding author: Daorong Wang
  • About author:
    Corresponding author: Wang Daorong, Email:
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引用本文：

史友权, 崇杨, 汤东, 熊清泉, 黄玉琴, 周怀成, 张琪, 金芝祥, 王道荣. sFRP2和Wnt/β-catenin通路在结直肠癌发生发展中的作用[J]. 中华结直肠疾病电子杂志, 2018, 07(06): 531-537.

Youquan Shi, Yang Chong, Dong Tang, Qingquan Xiong, Yuqin Huang, Huaicheng Zhou, Qi Zhang, Zhixiang Jin, Daorong Wang. The role of sFRP2 and Wnt /β-catenin pathway in the development and progression of colorectal cancer[J]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2018, 07(06): 531-537.

目的

研究分泌型卷曲蛋白2（Secreted Frizzled-related proteins 2，sFRP2）与Wnt/β-catenin通路在结直肠癌发生发展中的作用。

方法

首先对正常结直肠黏膜组织和结直肠癌组织行免疫组化实验，检测sFRP2及β-catenin的表达水平和阳性率。其次通过质粒转染，使得sFRP2在HCT116细胞中的表达上调，通过Western blot实验验证，然后行CCK-8实验、划痕实验、Transwell实验，分析sFRP2表达上调后对细胞增殖、迁移和侵袭的影响。

结果

正常结直肠黏膜组中sFRP2阳性表达率显著高于结直肠癌组（χ2=35.902，P=0.000）。相反，结直肠癌组中β-catenin膜表达缺失率和异位表达率显著高于正常结直肠黏膜组（χ2=23.149，P=0.000；χ2=27.002，P=0.000）。sFRP2阳性表达、β-catenin膜表达缺失、异位表达与肿瘤组织分化明显相关（χ2=5.420，P=0.020；χ2=6.472，P=0.011；χ2=7.158，P=0.007），而与性别、年龄、肿瘤部位、肿瘤大小、肿瘤分期和淋巴结转移无关（P>0.05）。sFRP2的阳性率与β-catenin的膜表达缺失率及异位表达率呈负相关，β-catenin的膜表达缺失率和异位表达率呈正相关。转染后sFRP2及β-catenin表达水平显著升高（t=25.430，P=0.001；t=15.000, P=0.001）。sFRP2转染组与对照组及空载质粒组相比，细胞增殖速度及迁移速度明显减慢（t=16.890, P=0.001；t=7.206，P=0.002）。与对照组相比，sFRP2转染组透膜细胞数明显少于对照组（t=25.459, P=0.001），细胞侵袭能力显著下降。

结论

sFRP2与Wnt/β-catenin通路相互作用，在结直肠癌的发生发展中起着重要作用。sFRP2的上调明显抑制了HCT116细胞的增殖、迁移和侵袭。

关键词: 结直肠肿瘤, sFRP2, β-catenin, HCT116细胞
Objective

To explore The role of sFRP2 and Wnt/β-catenin pathway in the development and progression of colorectal cancer.

Methods

Immunohistochemical staining was used to detect the expression of sFRP2 and β-catenin in colorectal cancer group and normal colorectal mucosa group, to detect the expression level and positive rate. The expression of sFRP2 gene in colorectal cancer cell line HCT116 was up-regulated by plasmid transfection, it was verified by Western blot. Then we conduct CCK-8 method, wound-healing assay and Transwell assay. Then the effects of up regulation of sFRP2 expression on cell proliferation, migration and invasion were analyzed.

Results

The results of immunohistochemistry showed that the positive rate of sFRP2 expression in normal colorectal mucosa is higher than colorectal cancer group (χ2=35.902, P=0.000); However, The rate of β-catenin membrane expression deficiency and ectopic expression in colorectal cancer group is higher than normal colorectal mucosa (χ2=23.149, P=0.000, χ2=27.002, P=0.000). The positive expression of sFRP2, the loss of expression and the ectopic expression of β-catenin membrane were significantly correlated with the differentiation of tumor tissues (χ2=5.420, P=0.020, χ2=6.472; P=0.011, χ2=7.158, P=0.007), however, it was not associated with sex, age, tumor location, tumor size, tumor stage and lymph node metastasis (P>0.05). The expression of sFRP2 was negatively correlated with β-catenin membrane expression deficiency and ectopic expression; There was a positive correlation between the β-catenin membrane expression deficiency and the ectopic expression. After plasmid transfection, the expression of sFRP2 and β-catenin significantly increased (t=25.430, P=0.001; t=15.000, P=0.001); The proliferation and migration rate of sFRP2 transfection group was significantly slower compared with the control group and the empty plasmid group (t=16.890, P=0.001; t=7.206, P=0.002); Compared with the control group, the number of transmembrane cells in sFRP2 transfected group was significantly fewer than that in the control group (t=25.459, P=0.001), and the cell invasion ability was significantly decreased.

Conclusion

The interaction between sFRP2 and Wnt/β-catenin pathway plays an important role in the development and progression of colorectal cancer. The up regulation of sFRP2 significantly inhibited the proliferation, migration and invasion of colorectal cancer cell line HCT116.

Key words: Colorectal neoplasms, sFRP2, β-catenin, HCT116
表1 sFRP2和β-catenin与结直肠癌临床特点的关系（例）
特征 例数 sFRP2阳性表达 β-catenin异常表达
膜表达缺失 异位表达
性别 ? ? ? ?
? 男 18 5 10 12
? 女 14 4 7 7
χ2值 ? 0.002 0.098 0.907
P值 ? 0.960 0.755 0.341
年龄 ? ? ? ?
? <70 25 7 13 15
? ≥70 7 2 4 4
χ2值 ? 0.001 0.058 0.019
P值 ? 0.976 0.810 0.892
肿瘤部位 ? ? ? ?
? 结肠 14 3 8 6
? 直肠 18 6 9 13
χ2值 ? 0.552 0.161 2.815
P值 ? 0.457 0.688 0.093
肿瘤大小 ? ? ? ?
? <5 cm 13 5 6 5
? ≥5 cm 19 4 11 14
χ2值 ? 1.157 0.427 3.970
P值 ? 0.282 0.513 0.071
肿瘤分期 ? ? ? ?
? Ⅰ/Ⅱ 17 6 10 11
? Ⅲ/Ⅳ 15 3 7 8
χ2值 ? 0.922 0.473 0.427
P值 ? 0.337 0.492 0.513
组织分化 ? ? ? ?
? 低分化 14 1 11 12
? 高中分化 18 8 6 7
χ2值 ? 5.420 6.472 7.158
P值 ? 0.020 0.011 0.007
淋巴结转移 ? ? ? ?
? 无 12 5 5 8
? 有 20 4 12 11
χ2值 ? 1.742 1.012 0.423
P值 ? 0.187 0.314 0.515
图1 sFRP2和β-catenin在结直肠组织中的的表达。1A：正常结肠组织（sFRP2）；1B：结直肠癌组（sFRP2）；1C：正常结肠组织（β-catenin）；1D：结直肠癌组（β-catenin）（免疫组化SP法×400）
表2 sFRP2及β-catenin在两组中的表达情况［例（%）］
组织类型 例数（例） sFRP2阳性表达 β-catenin异常表达
膜表达缺失 异位表达
正常结直肠黏膜组 32 32（100） 0（0） 0（0）
结直肠癌组 32 9（28.1） 17（53.1） 19（59.4）
P值 ? <0.001* <0.001* <0.001*
表3 结直肠癌中sFRP2和β-catenin表达的相关性（例）
分组 ? β-catenin膜表达缺失 β-catenin异位表达
? ? ＋ - ＋ -
sFRP2表达 ＋ 2 7 2 7
- 15 8 17 6
r值 ? -0.39 ? -0.47 ?
P值 ? 0.03 ? 0.01 ?
β-catenin异位表达 ＋ 14 5 ? ?
- 3 10 ? ?
r值 ? 0.50 ? ? ?
P值 ? 0.004 ? ? ?
图2 转染前后HCT116细胞中sFRP2、β-catenin的表达（sFRP2及β-catenin转染前后相比*P<0.05）
图3 sFRP2对结直肠癌HCT116细胞生长曲线的影响（sFRP2转染组与对照组相比*P<0.05）
图4 sFRP2对结直肠癌细胞HCT116细胞迁移能力的影响（sFRP2转染组与对照组相比*P<0.05）
图5 sFRP2对结直肠癌细胞HCT116细胞侵袭能力的影响（sFRP2转染组与对照组相比*P<0.05）
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