摘要
目的:研究白细胞介素-22(interleukin-22,IL-22)对胰腺癌细胞增殖、迁移和侵袭能力的影响并探索其可能机制。探索IL-22对不同恶性程度的胰腺癌细胞系促癌效果差异。方法:选择恶性程度较低的SW1990和恶性程度较高的CAPAN-2两种胰腺导管癌细胞做平行实验。MTT法检测IL-22对胰腺癌细胞增殖的影响;细胞划痕实验观察IL-22对胰腺癌细胞迁移能力的影响;Transwell小室侵袭实验检测IL-22对胰腺癌细胞侵袭能力的影响;RT-PCR检测经IL-22处理前后,胰腺癌细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)、高迁移率族蛋白B1(high mobility group protein B1,HMGB1)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的mRNA表达差异;Western blot实验检测IL-22处理前后,细胞中信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)、p-STAT3、VEGF、HMGB1、MMP9的蛋白表达差异。以上实验均进行两种胰腺癌细胞系之间的平行比较。结果:MTT实验中,在不同浓度的IL-22作用下,胰腺癌细胞光密度(optical density,OD)值随着浓度增加逐渐增高,CAPAN-2细胞的增长率大于SW1990细胞的增长率;划痕实验显示,在IL-22作用后24 h和48 h,划痕相对宽度分别逐渐减小,CAPAN-2细胞的24 h变化率和48 h变化率均大于SW1990细胞;Transwell实验显示,IL-22作用下,侵袭过膜细胞测得OD值较空白对照组增加,其中CAPAN-2细胞刺激前后增长率大于SW1990细胞;RT-PCR结果显示,IL-22可以显著增强VEGF、HMGB1、MMP9的mRNA表达水平,其中CAPAN-2细胞受IL-22刺激后三种促癌因子mRNA表达增强率均大于SW1990细胞;Western blot结果显示,IL-22可以明显增强STAT3通路磷酸化,STAT3蛋白表达下降,p-STAT3蛋白表达增加,其下游VEGF、HMGB1、MMP9的蛋白表达水平显著上升,其中CAPAN-2细胞的变化率大于SW1990细胞的变化率,以上实验结果差异均有统计学意义(P<0.05)。结论:IL-22通过磷酸化STAT3�
Objective:To investigate the effect of interleukin-22(IL-22)on the proliferation,migration and invasion of human pancreatic cancer cells and its mechanism.To explore the effect of IL-22 on pancreatic cancer cell lines with different differentiation degree.Methods:SW1990 and CAPAN-2 were choose as two kinds of pancreatic ductal carcinoma cells to do parallel experiments according to the degree of differentiation.MTT assay was used to detect the effect of IL-22 on the proliferation of two kinds of pancreatic cancer cells.Cell scratch assay was used to observe the effect of IL-22 on the migration ability of two pancreatic cancer cell lines.Transwell invasion assay was used to detectthe effect of IL-22 on the invasion ability.Detection of RT-PCR before and after the IL-22 treatment in cells was done,and it was about the difference of the VEGF(vascular endothelial growth factor),HMGB1(high mobility group protein B1),MMP9(matrix metalloproteinase 9)mRNA expression.Western blot assay before and after the IL-22 treatment,its about the difference of STAT3,p-STAT3,VEGF,HMGB1,MMP9 protein expression in the cells and parallel comparisons between the two pancreatic cancer cell lines were done above all the experiments.Results:In the MTT experiment,under the effects of IL-22 on the different concentration,OD values measured were the same with increasing concentration increased gradually,CAPAN-2 cell growth rate is greater than the growth rate of SW1990 cells.Scratch test results after 24 h and 48 h showed that the relative scratch width decreases by IL-22.The 24 h and 48 h change rate of CAPAN-2 cells was greater than the rate of SW1990 cells.Transwell results showed that,stimulated by IL-22,the OD value increased compared with blank control group,the growth rate of CAPAN-2 cells was greater than that of SW1990 cells.RT-PCR results showed that after IL-22 stimulation can significantly enhance the mRNA expression level of VEGF,HMGB1,MMP9,three kinds of cancer promoting factor mRNA expression increased more in CAPAN-2 than in SW
作者
华晔
葛春林
HUA Ye;GE Chunlin(Department of Urology,Shengjing Hospital of China Medical University,Liaoning Shenyang 110001,China;Department of Pancreas and Biliary Tract,the First Affiliated Hospital of China Medical University,Liaoning Shenyang 110001,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第23期4238-4243,共6页
Journal of Modern Oncology
基金
辽宁省教育厅科研立项(编号:L2014294)
辽宁省沈阳市科技项目(编号:F15-199-1-48)。
