天津医药

天津医药

• 细胞与分子生物学 • 上一篇    下一篇

TGF-β1对结直肠癌细胞株侵袭转移及IL-22表达的影响

阮洪训，秦晓宁，黄炜，李猛，赵晶，任鹏涛，郝英豪，林林△   

  1. 基金项目：河北省科技厅项目（162777202）；河北省卫生厅重点科技研究计划项目（20170564） 作者单位：河北医科大学第二医院肛肠外科（邮编050000） 作者简介：阮洪训（1981），男，硕士，主治医师，主要从事结直肠肿瘤研究 △通讯作者 E-mail: ll779242000@163.com
  • 出版日期:2019-09-15 发布日期:2019-09-15

The influence of TGF-β1 on cell invasiveness, cell metastasis and the expression of IL-22 in colorectal cancer cell line

RUAN Hong-xun, QIN Xiao-ning, HUANG Wei, LI Meng, ZHAO Jing, REN Peng-tao, HAO Ying-hao, LIN Lin△   

  1. Department of Anorectal Surgery, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China △Corresponding Author E-mail：ll779242000@163.com
  • Published:2019-09-15 Online:2019-09-15

PDF (PC)

5204

摘要： 目的 探讨转化生长因子β1（TGF-β1）对结直肠癌细胞株侵袭转移及白细胞介素22（IL-22）蛋白表达的 影响。方法 以结直肠癌HCT116细胞株为研究对象，以不同浓度（0、5、10、15、20、30、50 μg/L）TGF-β1处理24、48 h 后，CCK-8法检测结直肠癌HCT116细胞体外增殖情况；将结直肠癌HCT116细胞株分为对照组和10 μg/L TGF-β1 处理组。Transwell侵袭迁移实验和划痕实验检测TGF-β1对HCT116细胞侵袭迁移能力的影响；采用RT-PCR检测 TGF-β1对HCT116细胞TGF-β1、IL-22 mRNA表达的影响；免疫细胞化学染色和Western blot 检测HCT116细胞中 TGF-β1、IL-22、信号转导和转录激活因子3（STAT3）、E-钙黏蛋白（E-cadherin）表达水平。结果 CCK-8结果显示， 0、5、10、15、20、30、50 μg/L TGF-β1 溶液刺激 24、48 h 后，不同 TGF-β1 浓度处理组结直肠癌 HCT116 细胞光密度 （OD）值差异均无统计学意义（均P＞0.05）。10 μg/L TGF-β1处理HCT116细胞48 h后，细胞侵袭能力、迁移能力和 迁移距离均明显升高（均P＜0.01）；TGF-β1处理组TGF-β1 mRNA表达水平明显高于对照组，而IL-22 mRNA表达水 平明显低于对照组（均P＜0.01）；免疫细胞化学染色和Western blot结果显示，TGF-β1处理组TGF-β1、STAT3蛋白表 达水平明显高于对照组，而IL-22、E-cadherin蛋白表达水平明显低于对照组（均P＜0.05）。结论 TGF-β1可能促进 HCT116细胞侵袭和迁移，但对HCT116细胞的增殖影响不明显，结直肠癌HCT116细胞中TGF-β1可能抑制IL-22蛋 白的表达。

关键词: 结直肠肿瘤, 肿瘤侵润, 肿瘤转移, 转化生长因子β1, 白细胞介素22

Abstract: Objective To investigate the effect of transforming growth factor β 1 (TGF-β1) on invasion, metastasis and expression of interleukin 22 (IL-22) protein of colorectal cancer cell line. Methods The colorectal cancer HCT116 cells were treated with different concentrations of TGF-β1 (0, 5, 10, 15, 20, 30, 50 μg/L) for 24 and 48 hours. The hyperplasia of HCT116 cells was detected by CCK-8 assay. The colorectal cancer HCT116 cells were divided into control group and TGF- β1 treated group. Transwell invasion and migration assay and Wound-healing assay were used to detect the effect of TGF-β 1 on the invasion and migration of HCT116 cells. RT-PCR was used to detect the effect of TGF-β1 on the expressions of TGF - β1 and IL-22 mRNA in HCT116 cells. Immunocytochemistry and Western blot assay were used to detect the expressions of TGF-β1, IL-22, STAT3 and E-cadherin protein in HCT116 cells. Results CCK-8 results show that there were no significant differences in the optical density (OD) values of HCT116 cells between different TGF-β1 concentration treatment groups (0, 5, 10, 15, 20, 30, 50 μg/L) after stimulation with TGF-β1 solution for 24 and 48 hours (P＞0.05). The invasive ability, migration ability and migration distance of cells were significantly increased after treatment with TGF- β1 (10 μg/L) for 48 hours (P＜0.01). The expression of TGF-β1 mRNA was significantly higher but the expression level of IL- 22 mRNA was significantly lower in TGF - β1 treated group than those in the control group (P＜0.01). The results of immunocytochemistry and Western blot assay showed that the expressions of TGF-β1 and STAT3 protein were significantly higher in TGF-β1 treated group than those in the control group, and the expressions of IL-22 and E-cadherin protein were significantly lower in TGF-β1 treated group than those in control group (P＜0.05). Conclusion TGF-β1 may promote the invasion and migration of HCT116 cells, but the effect on the proliferation of HCT116 cells is not obvious. TGF-β1 may inhibit the expression of IL-22 protein in HCT116 cells.

Key words: colorectal neoplasms, neoplasm invasiveness, neoplasm metastasis, transforming growth factor beta1, interleukin-22