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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 ›› 2019, Vol. 08 ›› Issue (04) : 377 -382. doi: 10.3877/cma.j.issn.2095-3224.2019.04.010

所属专题： 文献；

论著

以IL-1β为基础的腹腔不同免疫水平对直肠癌术后腹腔微转移灶转归的作用
鲁兵1,†(), 张振宇1, 倪荔1, 严东羿1, 朱哲1, 高玮1, 袁彪1, 杨颻1, 傅传刚1   
  1. 1. 200120　上海，同济大学附属东方医院胃肠肛肠外科
  • 收稿日期:2018-01-23 出版日期:2019-08-25
  • 通信作者: 鲁兵
  • 基金资助:
    上海市卫计委局级项目(No.201440305); 上海市科委科研项目(No.19411966500)

Effect of IL-1β based intraperitoneal immunity levels on the prognosis of intraperitoneal micrometastasis after rectal cancer operation

Bing Lu1,†(), Zhenyu Zhang1, Li Ni1, Dongyu Yan1, Zhe Zhu1, Wei Gao1, Biao Yuan1, Yao Yang1, Chuangang Fu1   

  1. 1. Department of General Surgery, East Hospitail, Tongji University, Shanghai 200120, China
  • Received:2018-01-23 Published:2019-08-25
  • Corresponding author: Bing Lu
  • About author:
    Corresponding author: Lu Bing, Email,
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鲁兵, 张振宇, 倪荔, 严东羿, 朱哲, 高玮, 袁彪, 杨颻, 傅传刚. 以IL-1β为基础的腹腔不同免疫水平对直肠癌术后腹腔微转移灶转归的作用[J]. 中华结直肠疾病电子杂志, 2019, 08(04): 377-382.

Bing Lu, Zhenyu Zhang, Li Ni, Dongyu Yan, Zhe Zhu, Wei Gao, Biao Yuan, Yao Yang, Chuangang Fu. Effect of IL-1β based intraperitoneal immunity levels on the prognosis of intraperitoneal micrometastasis after rectal cancer operation[J]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2019, 08(04): 377-382.

目的

探讨IL-1β介导的腹腔内免疫功能变化对直肠癌腹腔微转移的影响。

方法

原代分离人腹膜间皮细胞，采用免疫荧光鉴定vimentin和VWF的表达，采用不同浓度的IL-1β和不同浓度的IL-1β抗体刺激共培养的人腹膜间质内皮细胞和癌细胞，流式细胞数检测细胞的粘附率。

结果

免疫荧光鉴定得出：vimentin为阳性表达，VWF为阴性表达，随着IL-1β浓度的增加，细胞粘附率依次增加；当加入抗体IL-1β后，随着抗体浓度的增加，人腹膜间质内皮细胞与癌细胞粘附率呈现下降趋势。且5 ng/mL的IL-1β含量时，细胞粘附率最高，显著高于其他组（P<0.05）；10 ng/mL的IL-1β含量时粘附率高于其他组，仅低于5 ng/mL的IL-1β组；加入抗体IL-1β后所有组别的粘附率趋势与不加前一致，但是所有组别的粘附率均降低，1 ng/mL IL-1β组＋0.1ug/mL抗体组和5 ng/mL IL-1β组＋0.5 ug/mL抗体组的细胞粘附率最高，阴性对照组细胞粘附率最低，显著低于其他组（P<0.05）。

结论

本研究显示IL-1β能够促进直肠癌细胞与腹膜间皮细胞粘附，并呈现浓度依耐性，提示其可能参与直肠癌腹膜转移。

关键词: 直肠肿瘤, 微转移, 免疫功能, IL-1β, IL-1β抗体
Objective

To investigate the effect of L-1β-mediated intraperitoneal immune function on peritoneal micrometastasis in rectal cancer.

Methods

Primary human peritoneal mesothelial cells were isolated and the expressions of vimentin and VWF were detected by immunofluorescence. Human peritoneal mesothelial cells and cancer cells were stimulated with different concentrations of IL-1β and IL-1β antibody. The rate of cell adhesion was detected by flow cytometry.

Results

The results of immunofluorescence showed that vimentin was positive and VWF was negative. With the increase of IL-1β, cell adhesion rate were increasing. When IL-1β was added, with the increase of antibody concentration, human peritoneal interstitial endothelial cells and cancer cell adhesion rate showed declining trend. When the concentration of IL-1β was 5 ng/mL, the cell adhesion rate was the highest, which was significantly higher than that of other groups (P<0.05). The adhesion rate trend of all the groups after adding IL-1β antibody was the same as that of the other groups, but the adhesion rate of all groups was decreased. The adhesion rate of 1 ng/mL IL-1β+ 0.1 ug/mL and 5 ng/mL IL-1β group+ 0.5 ug/mL antibody group were the highest, and the lowest in the negative control group (P<0.05).

Conclusion

Our results show that IL-1β promotes the adhesion of rectal cancer cells to peritoneal interstitial endothelial cells in a concentration-dependent manner, indicating a favorable role of IL-1β in peritoneal carcinomatosis.

Key words: Colorectal neoplasms, Immunity, Micrometastasis, IL-1β, IL-1βantibody
图1 人腹膜间皮细胞原代培养照片。1A：第1天，1B：第3天，1C：第5天，1D：第9天，1E：P1代，1F：P2代（×100倍）
图2 原代培养人腹膜间皮细胞鉴定结果。2A：DAPI染色，2B：Vimentin染色，2C：Merge，2D：DAPI染色，2E：VWF染色，2F：Merge （×100倍）
图3 各实验组流式细胞检测结果。3A：阴性对照组，3B：0 ng/mL IL-1β组，3C：1 ng/mL IL-1β组，3D：5 ng/mL IL-1β组，3E：10 ng/mL IL-1β组，3F：0 ng/mL IL-1β＋0 μg/mL抗体组，3G：1 ng/mL IL-1β＋0.1 μg/mL抗体组，3H：5 ng/mL IL-1β＋0.5 μg/mL抗体组，3I：5 ng/mL IL-1β＋1 μg/mL抗体组
图4 各实验组细胞的粘附率结果
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