沉默白细胞介素17 受体B基因对肺腺癌A549细胞增殖、侵袭和凋亡的影响及其机制

doi:10.13481/j.1671-587X.20260316

摘要/Abstract

摘要：

目的探讨白细胞介素17受体B（IL-17RB）基因沉默对肺腺癌A549细胞增殖、凋亡和侵袭的影响，并阐明其在肺腺癌发生发展中的作用。方法培养正常人支气管上皮BEAS-2B细胞和人肺腺癌A549细胞，采用实时荧光定量PCR（RT-qPCR）法和Western blotting法检测2种细胞中IL-17RB mRNA和蛋白表达水平。采用脂质体法将IL-17RB小干扰RNA（si-RNA）片段转染至A549细胞中，细胞分为si-NC组和si-IL-17RB组。采用RT-qPCR法和Western blotting法检测IL-17RB沉默效果，细胞计数试剂盒8（CCK-8）法检测2组A549细胞增殖活性，5-乙炔基-2'-脱氧尿嘧啶核苷（EdU）染色检测2组A549细胞中EdU阳性细胞率，流式细胞术检测2组A549细胞凋亡率，Transwell小室实验检测2组A549细胞的迁移细胞数和侵袭细胞数，Western blotting法检测2组A549细胞中B细胞淋巴瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）、基质金属蛋白酶9（MMP-9）、N-钙黏蛋白（N-cadherin）、波形蛋白（Vimentin）、E-钙黏蛋白（E-cadherin）、Snail同源物（Snail）和磷酸化P65（p-P65）蛋白表达水平。结果 RT-qPCR法和Western blotting法，与BEAS-2B细胞比较，A549细胞中IL-17RB mRNA和蛋白表达水平明显升高（P<0.01）；与si-NC组比较，si-IL-17RB组A549细胞中IL-17RB mRNA和蛋白表达水平均明显降低（P<0.01）。CCK-8法和EdU染色，与si-NC组比较，si-IL-17RB组A549细胞增殖活性和A549细胞中EdU阳性细胞率明显降低（P<0.01）。流式细胞术，与si-NC组比较，si-IL-17RB组A549细胞中细胞凋亡率明显升高（P<0.01）。Transwell小室实验，与si-NC组比较，si-IL-17RB组A549细胞的迁移细胞数和侵袭细胞数均明显减少（P<0.01）。Western blotting法，与si-NC组比较，si-IL-17RB组A549细胞中Bcl-2、Vimentin、N-cadherin、MMP-9、Snail和p-P65蛋白表达水平明显降低（P<0.01），Bax和E-cadherin蛋白表达水平明显升高（P<0.01）。结论沉默IL-17RB基因可降低肺腺癌A549细胞增殖和侵袭能力，促进细胞凋亡，并抑制细胞中p-P65蛋白表达水平，其作用机制可能与抑制核因子κB（NF-κB）信号通路有关。

关键词: 白细胞介素17受体B, 肺腺癌, 细胞增殖, 细胞凋亡, 上皮-间质转化

Abstract:

Objective To discuss the effect of interleukin-17 receptor B （IL-17RB） gene silencing on the proliferation， apoptosis and invasion of lung adenocarcinoma A549 cells， and to clarify its role in the occurrence and development of lung adenocarcinoma. Methods The normal human bronchial epithelial BEAS-2B cells and human lung adenocarcinoma A549 cells were cultured； real-time fluorescence quantitative PCR（RT-qPCR） and Western blotting method were used to detect the expression levels of IL-17RB mRNA and protein in the two kinds of cells. Liposome method was used to transfect IL-17RB small interfering RNA （si-RNA） fragments into the A549 cells， and the cells were divided into si-NC group and si-IL-17RB group. RT-qPCR and Western blotting methods were used to detect the silencing effect of IL-17RB； CCK-8 method was used to detect the proliferation activities of A549 cells in two groups； 5-ethynyl-2'-deoxyuridine （EdU） staining was used to detect the EdU positive cell rates of A549 cells in two groups； flow cytometry was used to detect the apoptotic rates of A549 cells in two groups； Transwell chamber assay was used to detect the numbers of migration cells and invasion cells of A549 cells in two groups； Western blotting method was used to detect the expression levels of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， matrix metalloproteinase 9 （MMP-9）， N-cadherin， Vimentin， E-cadherin， Snail homolog （Snail） and phosphorylated P65 （p-P65） proteins in A549 cells in two groups. Results The RT-PCR results showed that compared with BEAS-2B cells， the expression levels of IL-17RB mRNA and protein in A549 cells were significantly increased （P<0.01）； compared with si-NC group， the expression levels of IL-17RB mRNA and protein in A549 cells in si-IL-17RB group were significantly decreased （P<0.01）. The CCK-8 assay results showed that compared with si-NC group， the proliferation activity of A549 cells in si-IL-17RB group was significantly decreased （P<0.01）. The EdU staining results showed that compared with si-NC group， the EdU positive cell rate of A549 cells in si-IL-17RB group was significantly decreased （P<0.01）. The flow cytometry results showed that compared with si-NC group， the apoptotic rate of A549 cells in si-IL-17RB group was significantly increased （P<0.01）. The Transwell chamber assay results showed that compared with si-NC group， the numbers of migration cells and invasion cells of A549 cells in si-IL-17RB group were significantly decreased （P<0.01）. The Western blotting results showed that compared with si-NC group， the expression levels of Bcl-2， Vimentin， N-cadherin， MMP-9， Snail and p-P65 proteins in A549 cells in si-IL-17RB group were significantly decreased （P<0.01）， while the expression levels of Bax and E-cadherin proteins were significantly increased （P<0.01）. Conclusion Silencing IL-17RB gene can reduce the proliferation and invasion abilities of lung adenocarcinoma A549 cells， promote cell apoptosis， and inhibit the expression level of p-P65 protein in cells； its mechanism may be related kinhibiting the nuclear factor-kappa B （NF-κB） singal pathway.

Key words: Interleukin-17 receptor B, Lung adenocarcinoma, Cell proliferation, Cell apoptosis, Epithelial-mesenchymal transition

中图分类号:

R734.2

任俏同,吴维华,王星翔,王世超,程一鹏,历春. 沉默白细胞介素17 受体B基因对肺腺癌A549细胞增殖、侵袭和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2026, 52(3): 738-745.

Qiaotong REN,Weihua WU,Xingxiang WANG,Shichao WANG,Yipeng CHENG,Chun LI. Effect of interleukin-17 recepter B gene silencing on proliferation, invasion and apoptosis of lung adenocarcinoma A549 cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 738-745.

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