切换至 "中华医学电子期刊资源库"

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:
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/OL]. 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/OL]. 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。

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.

表1 sFRP2和β-catenin与结直肠癌临床特点的关系(例)
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(%)]
表3 结直肠癌中sFRP2和β-catenin表达的相关性(例)
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)
[1]
刘坤, 赵任.结直肠癌肝转移标志物的研究进展[J]. 中华胃肠外科杂志, 2013, 16(8):794-796.
[2]
Bryan T. MacDonald, Xi He. Frizzled and LRp5/6 Receptors for Wnt/β-Catenin Signaling [J]. Cold Spring Harbor perspectives in Biology, 2012, 4(12): S107.
[3]
Niehrs C. The complex world of Wnt receptor signalling [J]. Nature Reviews Molecular Cell Biology, 2012, 13(12): 767.
[4]
Maruyama K, Ochiai A, Akimoto S, et al. Cytoplasmic beta-catenin accumulation as a predictor of hematogenous metastasis in human colorectal cancer [J]. Oncology, 2000, 59(59): 302-309.
[5]
Oberwalder M, Zitt M, Wöntner C, et al. SFRP2 methylation in fecal DNA-a marker for colorectal polyps [J]. International Journal of Colorectal Disease, 2008, 23(1): 15.
[6]
Mazieres J, He B, You L, et al. Wnt signaling in lung cancer [J]. Cancer Letters, 2005, 222(1): 1-10.
[7]
Sparks AB, Morin PJ, Vogelstein B, et al. Mutational Analysis of the ApC/β-Catenin/Tcf pathway in Colorectal Cancer [J]. Cancer Research, 1998, 58(6): 1130-1134.
[8]
刘宁, 姜海行, 黄振宁, 等. SFRP2和β-catenin在大肠癌中的表达及其临床意义[J]. 世界华人消化杂志, 2008, 16(35): 3963-3969.
[9]
Bafico A, Liu G, Goldin L, et al. An autocrine mechanism for constitutive Wnt pathway activation in human cancer cells [J]. Cancer Cell, 2004, 6(5): 497.
[10]
Suzuki H, Gabrielson E, Chen W, et al. A genomic screen for genes upregulated by demethylation and histone deacetylase inhibition in human colorectal cancer [J]. Nature Genetics, 2002, 31(2): 141.
[11]
Suzuki H, Toyota M, Nojima M, et al. SFRP, a family of new colorectal tumor suppressor candidate genes [J]. Nihon Rinsho Japanese Journal of Clinical Medicine, 2005, 63(4): 707-719.
[12]
Hans Clevers, Roel Nusse. Wnt/β-Catenin Signaling and Disease [J]. Cell, 2012, 149(6): 1192.
[13]
Willert K, Nusse R. Wnt proteins [J]. Cold Spring Harbor perspectives in Biology, 2012, 4(4): a007864.
[14]
Wang J, Sinha T, Wynshaw-Boris A. Wnt signaling in mammalian development: lessons from mouse genetics [J]. Cold Spring Harbor perspectives in Biology, 2012, 4(5): 570-581.
[15]
Anastas JN, Moon RT. Wnt signalling pathways as therapeutic targets in cancer [J]. Nature Reviews Cancer, 2013, 13(1): 11-26.
[16]
Reya T, Clevers H. Wnt signalling in stem cells and cancer [J]. Nature, 2005, 434(7035): 843-850.
[17]
Bienz M, Clevers H. Linking colorectal cancer to Wnt signaling [J]. Cell, 2000, 103(2): 311.
[18]
马思平, 单吉贤, 王辉. E-cadherin和β-catenin在大肠癌组织中的表达及与预后的关系[J]. 实用肿瘤学杂志, 2003, 17(4): 268-270.
[19]
鱼海峰, 王道荣, 张云, 等. SFRP2基因超甲基化与结直肠癌临床病理关系的研究[J]. 国际外科学杂志, 2012, 39(8): 526-529.
[20]
潘世杰. SFRP2和β-catenin在前列腺癌中的表达和意义[D]. 福州: 福建医科大学, 2014.
[21]
肖灿. SFRP2基因甲基化促进口腔粘膜鳞癌发生的分子机制研究[D]. 苏州: 苏州大学, 2013.
[22]
Tung JN. CyclinD1 protein expressed in pterygia is associated with β-catenin protein localization [J]. Molecular Vision, 2010, 16(291-97): 2733.
[23]
Shirali S, Aghaei M, Shabani M, et al. Adenosine induces cell cycle arrest and apoptosis via cyclinD1/Cdk4 and Bcl-2/Bax pathways in human ovarian cancer cell line OVCAR-3 [J]. Tumor Biology, 2013, 34(2): 1085-1095.
[24]
Artegiani B, Lindemann D, Calegari F. Overexpression of cdk4 and cyclinD1 triggers greater expansion of neural stem cells in the adult mouse brain [J]. Journal of Experimental Medicine, 2011, 208(5): 937-948.
[1] 罗青杉, 梅海涛, 郝家领, 蔡锦锋, 周润楷, 温玉刚. 连接蛋白43通过调控细胞周期抑制结直肠癌的增殖机制研究[J/OL]. 中华普通外科学文献(电子版), 2024, 18(05): 344-349.
[2] 徐逸男. 不同术式治疗梗阻性左半结直肠癌的疗效观察[J/OL]. 中华普外科手术学杂志(电子版), 2025, 19(01): 72-75.
[3] 韩加刚, 王振军. 梗阻性左半结肠癌的治疗策略[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(06): 450-458.
[4] 梁轩豪, 李小荣, 李亮, 林昌伟. 肠梗阻支架置入术联合新辅助化疗治疗结直肠癌急性肠梗阻的疗效及其预后的Meta 分析[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(06): 472-482.
[5] 严虹霞, 王晓娟, 张毅勋. 2 型糖尿病对结直肠癌患者肿瘤标记物、临床病理及预后的影响[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(06): 483-487.
[6] 赵磊, 刘文志, 林峰, 于剑, 孙铭骏, 崔佑刚, 张旭, 衣宇鹏, 于宝胜, 冯宁. 深部热疗在改善结直肠癌术后辅助化疗副反应及生活质量中的作用研究[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(06): 488-493.
[7] 黄海洋, 邝永龙, 陈嘉胜. 基层医院结直肠肿瘤经自然腔道取标本手术30 例分析[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(06): 510-518.
[8] 张蔚林, 王哲学, 白峻阁, 黄忠诚, 肖志刚. 利用TCGA数据库构建基于miRNA的结直肠癌列线图预后模型[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(05): 381-388.
[9] 张伟伟, 陈启, 翁和语, 黄亮. 随机森林模型预测T1 期结直肠癌淋巴结转移的初步研究[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(05): 389-393.
[10] 任佳琪, 刁德昌, 何自衍, 张雪阳, 唐新, 李文娟, 李洪明, 卢新泉, 易小江. 网膜融合线导向的脾曲游离技术在左半结肠癌根治术中的应用[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(05): 362-367.
[11] 张迪, 王春霞, 张学东, 李发馨, 庞淅文, 陈一锋, 张维胜, 王涛. 梗阻性左半结直肠癌自膨式金属支架置入后行腹腔镜手术与开腹手术的短期临床疗效比较[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(05): 375-380.
[12] 季鹏程, 鄂一民, 陆晨, 喻春钊. 循环外泌体相关生物标志物在结直肠癌诊断中的研究进展[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(04): 265-273.
[13] 李佳莹, 王旭丹, 梁雪, 张雷, 李佳英. 1990~2021年中国结直肠癌死亡趋势分析[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(04): 274-279.
[14] 戈伟, 陈刚. 纳米炭导航行淋巴示踪在结直肠癌TNM分期中淋巴分期价值的临床研究[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(04): 288-293.
[15] 崔精, 鲍一帆, 沈晓明, 杨增辉, 高森, 鲍传庆. 结直肠癌中circMFSD12对肿瘤细胞功能及5-FU敏感性的调控[J/OL]. 中华结直肠疾病电子杂志, 2024, 13(04): 294-302.
阅读次数
全文


摘要