国际肿瘤学杂志 ›› 2022, Vol. 49 ›› Issue (2): 73-78.doi: 10.3760/cma.j.cn371439-20210526-00011

• 论著 • 上一篇    下一篇

外源性AGR2对结肠癌细胞增殖及侵袭能力的影响

来比江·吾斯曼, 曹博威, 张文斌(), 高华()   

  1. 新疆医科大学第一附属医院胃肠外科,乌鲁木齐 830054
  • 收稿日期:2021-05-26 修回日期:2021-09-20 出版日期:2022-02-08 发布日期:2022-03-11
  • 通讯作者: 张文斌,高华 E-mail:zwb3216@sina.com;drgaoh@163.com
  • 基金资助:
    新疆维吾尔自治区卫生健康委青年科技创新基金(WJWY-202149)

Effects of exogenous AGR2 on the proliferation and invasion abilities of colon cancer cells

Laibijiang Wusiman, Cao Bowei, Zhang Wenbin(), Gao Hua()   

  1. Department of Gastrointestinal Surgery, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China
  • Received:2021-05-26 Revised:2021-09-20 Online:2022-02-08 Published:2022-03-11
  • Contact: Zhang Wenbin,Gao Hua E-mail:zwb3216@sina.com;drgaoh@163.com
  • Supported by:
    Xinjiang Uygur Autonomous Region Health Commission Youth Science and Technology Innovation Project(WJWY-202149)

摘要:

目的 检测3种结肠癌细胞株SW480、SW620和COLO205培养液中前梯度蛋白2(AGR2)的表达情况,并探讨不同浓度外源性AGR2对SW620细胞增殖、迁移和侵袭能力的影响。方法 采用蛋白质印迹法检测结肠癌细胞SW480、SW620和COLO205培养液中AGR2蛋白表达水平。将SW620细胞分为空白对照组、前梯度蛋白2同源人重组蛋白(rAGR2)低浓度组(100 μg/ml)和rAGR2高浓度组(200 μg/ml),采用CCK-8实验、细胞划痕实验、Transwell迁移和侵袭实验检测不同浓度rAGR2对SW620细胞生物学行为的影响。结果 蛋白质印迹法检测结果显示,SW480、SW620、COLO205细胞AGR2蛋白表达水平分别为0.545±0.097、0.662±0.040、0.882±0.156,差异有统计学意义(F=7.46,P=0.024),COLO205细胞株AGR2蛋白水平明显高于SW480、SW620细胞株(P=0.009;P=0.047)。CCK-8实验结果显示,空白对照组、rAGR2低浓度组、rAGR2高浓度组SW620细胞增殖活性分别为0.422±0.031、0.542±0.040、0.574±0.033,差异有统计学意义(F=26.35,P<0.001),rAGR2低浓度组、rAGR2高浓度组明显高于空白对照组(均P<0.001)。细胞划痕实验结果显示,3组细胞36 h细胞划痕面积百分比分别为(28.029±2.107)%、(20.642±0.983)%、(16.951±1.608)%,差异有统计学意义(F=35.85,P<0.001),rAGR2低浓度组高于空白对照组(P=0.001),rAGR2高浓度组高于rAGR2低浓度组(P=0.032)。细胞迁移实验结果显示,3组细胞迁移数分别为(447.1±32.3)个、(513.1±55.8)个、(632.4±50.3)个,差异有统计学意义(F=35.62,P<0.001),rAGR2低浓度组多于空白对照组(P=0.007),rAGR2高浓度组多于rAGR2低浓度组(P<0.001)。侵袭实验结果显示,3组细胞侵袭数分别为(369.1±56.1)个、(505.1±34.4)个、(579.0±71.5)个,差异有统计学意义(F=32.40,P<0.001),rAGR2低浓度组多于空白对照组(P<0.001),rAGR2高浓度组多于rAGR2低浓度组(P=0.010)。结论 不同侵袭性结肠癌细胞的细胞外液中AGR2蛋白表达量不同,随着侵袭能力的增高,AGR2蛋白的表达也增高。AGR2蛋白可提高结肠癌细胞SW620增殖、迁移和侵袭能力。

关键词: 结肠肿瘤, 细胞增殖, 细胞运动, AGR2

Abstract:

Objective To detect the expressions of anterior gradient protein 2 (AGR2) in the cultures of three colon cancer cell lines SW480, SW620 and COLO205, and to investigate the effects of different concentrations of exogenous AGR2 on the proliferation, migration and invasion abilities of SW620 cells. Methods Western blotting was used to detect the expression levels of AGR2 protein in SW480, SW620 and COLO205 colon cancer cell cultures. SW620 cells were divided into blank control group, anterior gradient protein 2 homologous human recombinant protein (rAGR2) low concentration group (100 μg/ml) and rAGR2 high concentration group (200 μg/ml), and CCK-8 assay, cell scratch assay and Transwell migration and invasion assay were used to detect the effects of different concentrations of rAGR2 on the biological behaviors of SW620 cells. Results Western blotting results showed that the expression levels of AGR2 protein in SW480, SW620 and COLO205 cells were 0.545±0.097, 0.662±0.040 and 0.882±0.156 respectively, with a statistically significant difference (F=7.46, P=0.024). The level of AGR2 protein in COLO205 cell line was significantly higher than that in SW480 and SW620 cell lines (P=0.009; P=0.047). The results of CCK-8 experiment showed that the proliferative activities of SW620 cells in the blank control group, rAGR2 low concentration group and rAGR2 high concentration group were 0.422±0.031, 0.542±0.040 and 0.574±0.033 respectively, with a statistically significant difference (F=26.35, P<0.001), and the rAGR2 low concentration group and rAGR2 high concentration group were significantly higher than the blank control group (both P<0.001). The results of cell scratching assay showed that the percentage of 36 h cell scratching area was (28.029±2.107)%, (20.642±0.983)% and (16.951±1.608)% for the three groups of cells respectively, with a statistically significant difference (F=35.85, P<0.001), the rAGR2 low concentration group was higher than the blank control group (P=0.001), and the rAGR2 high concentration group was higher than the rAGR2 low concentration group (P=0.032). The results of cell migration assay showed that the number of cells migrated in the three groups was 447.1±32.3, 513.1±55.8 and 632.4±50.3 respectively, with a statistically significant difference (F=35.62, P<0.001), the rAGR2 low concentration group was more than the blank control group (P=0.007), and the rAGR2 high concentration group was more than the rAGR2 low concentration group (P<0.001). The results of the invasion assay showed that the number of cells invaded in the three groups was 369.1±56.1, 505.1±34.4 and 579.0±71.5 respectively, with a statistically significant difference (F=32.40, P<0.001), the rAGR2 low concentration group was more than the blank control group (P<0.001), and the rAGR2 high concentration group was more than the rAGR2 low concentration group (P=0.010). Conclusion The expression of AGR2 protein varies in the extracellular fluid of different invasive colon cancer cells and increases with the invasive ability. AGR2 protein can increase the proliferation, migration and invasive abilities of colon cancer cells SW620.

Key words: Colonic neoplasms, Cell proliferation, Cell movement, AGR2