吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (6): 1510-1517.doi: 10.13481/j.1671-587X.20210622

• 临床研究 • 上一篇    下一篇

重组人IL-17A在胃癌组织中的表达及其对胃癌BGC-823细胞增殖、侵袭、迁移和凋亡的影响

母润红1,2,艾一玖1,李雨澎2,林睿3,叶思萍3,马方4,郭笑1,3()   

  1. 1.北华大学附属医院病理科,吉林 吉林 132013
    2.北华大学基础医学院免疫学教研室,吉林 吉林 132013
    3.北华大学药学院药学实验室,吉林 吉林 132013
    4.吉化集团公司总医院病理科,吉林 吉林 132013
  • 收稿日期:2021-04-23 出版日期:2021-11-28 发布日期:2021-12-14
  • 通讯作者: 郭笑 E-mail:395798544@qq.com
  • 作者简介:母润红(1975-),女,吉林省吉林市人,副教授,医学博士,主要从事肿瘤免疫学方面的研究。
  • 基金资助:
    吉林省科技厅科技发展计划项目(20190303047SF);吉林省卫建委卫生与健康创新项目(2020J024)

Expression of recombinant human IL-17A in gastric cancer tissue and its effects on proliferation, invasion, migration and apoptosis of gastric cancer BGC-823 cells

Runhong MU1,2,Yijiu AI1,Yupeng LI2,Rui LIN3,Siping YE3,Fang MA4,Xiao GUO1,3()   

  1. 1.Department of Pathology,Affiliated Hospital,Beihua University,Jilin 132013,China
    2.Department of Immunology,School of Basic Medical Science,Beihua University,Jilin 132013,China
    3.Department of Pharmacology,School of Pharmacology,Beihua University,Jilin 132013,China
    4.Department of Pathology,General Hospital,Jihua Group Company,Jilin 132011,China
  • Received:2021-04-23 Online:2021-11-28 Published:2021-12-14
  • Contact: Xiao GUO E-mail:395798544@qq.com

摘要: 目的

检测人白细胞介素17A(IL-17A)重组蛋白在人胃癌组织中的表达及其对胃癌细胞增殖、侵袭、迁移和凋亡的影响,并初步探讨其作用机制。

方法

收集胃癌患者术后石蜡标本30例、肠上皮化生标本20例和正常胃组织标本10例。免疫组织化学染色法检测各种组织中IL-17A和白细胞介素17受体A(IL-17RA)的阳性表达率, Pearson相关分析法分析IL-17A与IL-17RA表达的相关性。将对数生长期BGC-823细胞分为对照组和不同浓度(50、100和200 μg·L-1)IL-17A组。MTT法检测各组胃癌BGC- 823细胞增殖活性,流式细胞术检测各组胃癌BGC-823细胞凋亡率,Transwell 实验检测各组胃癌BGC-823细胞侵袭细胞数,划痕实验检测各组胃癌BGC-823细胞迁移率, Western blotting法检测各组胃癌BGC-823细胞中血管内皮生长因子(VEGF)、基质金属蛋白酶9(MMP-9)、B细胞淋巴瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平。

结果

肠上皮化生和胃癌组织中IL-17A和IL-17RA阳性表达率明显高于正常胃组织(P<0.01),胃癌组织中IL-17A与IL-17RA表达呈正相关关系(r=0.891,P<0.01)。IL-17A作用48和72 h后,与对照组比较,100和200 μg·L-1 IL-17A组胃癌BGC-823细胞增殖活性升高(P<0.05)。与对照组比较,100和200 μg·L-1 IL-17A组胃癌BGC-823细胞凋亡率明显降低(P<0.01),侵袭细胞数和细胞迁移率升高(P<0.05);Western blotting法检测,与对照组比较,200 μg·L-1 IL-17A组胃癌BGC-823细胞中VEGF、MMP-9和Bcl-2蛋白表达水平明显升高(P<0.05),Bax蛋白表达水平明显降低(P<0.05)。

结论

重组人IL-17A可促进胃癌BGC-823细胞增殖、迁移和侵袭,抑制细胞凋亡,其作用机制可能与促进侵袭相关蛋白VEGF和MMP-9表达以及抑制凋亡相关蛋白Bcl-2和Bax表达有关。

关键词: 白细胞介素17A, 白细胞介素17受体A, 胃肿瘤, 细胞凋亡, 细胞迁移

Abstract: Objective

To detect the expression of human interleukin-17A (IL-17A) recombinant protein in the human gastric cancer tissue and its effects on the proliferation, invasion, migration and apoptosis of gastric cancer cells, and to explore their mechanisms.

Methods

Thirty cases of postoperative paraffin specimens of the patients with gastric cancer,20 cases of intestinal metaplasia specimens, and 10 cases of normal gastric tissue were collected. The positive expression rates of IL-17A and interleukin-17 receptor A (IL-17RA) in different kinds of tissues were detected by immunohistochemical staining.The correlation between the expressions of IL-17A and IL-17RA was analyzed by Pearson correlation analysis.The BGC-823 cells in logarithmic growth phase were divided into control group and different concentrations (50,100, and 200 μg·L-1) of IL-17A groups. The proliferation activities of the gastric cancer BGC-823 cells in various groups were detected by MTT method. The numbers of invasion cells of the gastric cancer BGC-823 cells in various groups were detected by Transwell test. The migration rates of the gastric cancer BGC-823 cells in various groups were detected by scratch test, and the expression levels of vascular endothelial growth factor (VEGF),matrix metalloproteinase-9 (MMP-9),B cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax) in the gastric cancer BGC-823 cells in various groups were detected by Western blotting method.

Results

The positive expression rates of IL-17A and IL-17RA in the intestinal metaplasia and gastric cancer tissue were significantly higher than those in normal gastric tissue(P<0.01). There was positive correlation between IL-17A expression and IL-17RA expression in the gastric cancer tissue(r=0.891,P<0.01). Compared with control group,the proliferation activities of the gastric cancer BGC-823 cells in 100 and 200 μg·L-1 IL-17A groups were significantly increased after treated with IL-17A for 48 and 72 h (P<0.05). Compared with control group, the apoptotic rates of gastric cancer BGC-823 cells in 100 and 200 μg·L-1 IL-17A groups were decreased significantly (P<0.01), the numbers of invasion cells and migration rates were increased (P<0.05).The Western blotting method results showed that compared with control group, the expression levels of VEGF, MMP-9 and Bcl-2 proteins in the gastric cancer BGC-823 cells in 200 μg·L-1 IL-17A group were increased (P<0.05), and the expression level of Bax protein was decreased(P<0.05).

Conclusion

The recombinant human IL-17A can promote the proliferation, migration, and invasion of the gastric cancer BGC-823 cells and inhibits the apoptosis,and its mechanism may be related to the promotion of expressions of invasion-related proteins VEGF, MMP-9, and the inhibition of apoptosis-related proteins Bcl-2 and Bax.

Key words: interleukin-17A, interleukin-17 receptor A, stomach neoplasms, apoptosis, cell migration

中图分类号: 

  • R735.2