Abstract:

Objective　Based on toll-like receptor 4(TLR4) and nuclear factor-kappa B (NF-κB), to explore the effect of long non-coding RNA (lncRNA) and maternal expression gene 3 (MEG3) on migration and invasion of pancreatic cancer cells. Methods　This study was conducted from January 2022 to January 2023. The human pancreatic cancer cell lines were divided into a blank group (no transfected PANC-1 cells), an idle group (transfected pCDHEF1-copGFP pCDH-Scramble with PANC-1), and an MEG3 group (PANC-1 transfection PCDH-EF1-MEG3-COPGFP pCDH-MEG3). The quantitative real-time polymerase chain reaction (QRT-PCR) was used to detect the expressions of lncRNA-MEG3 and TLR4/NF-κB in the pancreatic cancer cells. The clonal formation rate was detected by the clonal assay. The apoptosis rate of the PANC-1 cells was detected by the flow cytometry. The invasion number and migration distance of the pancreatic cancer cells were detected by the Transwell cell and scratch assay. The expression levels of TLR4, NF-κB, NF-κBp65, and myeloid differentiation factor 88 (MyD88) in the pancreatic cancer cells were detected by the Western blot. t test, one-way analysis of variance, and SNK-q test were used for the statistical analysis. Results　The clonal formation rate, invasion number, and migration distance of the PANC-1 cells in the MEG3 group were lower than those in the blank group and the idle group, and the apoptosis rate of the PANC-1 cells was higher, with statistical difference (F=23.070, 99.390, 23.980, and 265.000; all P<0.05). The protein expressions of TLR4, NF-κB, NF-κBp65, and MyD88 of PANC-1 cells in the MEG3 group and the inhibitor group were lower than those in the blank group and the idle group (all P<0.05). Conclusion　LncRNA-MEG3 can regulate the migration and invasion of PANC-1 cells inhibiting the expression of TLR4/NF-κB gene.

Key words:

Pancreatic cancer, Transfection, Invasion, Cloning, Apoptosis

摘要：

目的　通过Toll样受体4（TLR4）及转录因子-κB（NF-κB）信号探究长链非编码RNA（lncRNA）-母系表达基因3（MEG3）对胰腺癌细胞迁移、侵袭的影响。方法　研究时间：2022年1月至2023年1月。将PANC-1（人胰腺癌细胞株）分为空白组（无转染PANC-1细胞），空转组（PANC-1转染pCDHEF1-copGFP pCDH-Scramble），MEG3组（PANC-1转染pCDH-EF1-MEG3–copGFP pCDH-MEG3）。通过实时荧光定量聚合酶链反应（qRT-PCR）检测胰腺癌细胞中lncRNA-MEG3、TLR4/NF-κB表达量，克隆法形成实验检测克隆形成率，流式细胞仪检测PANC-1细胞凋亡率，Transwell小室及划痕实验分别检测胰腺癌细胞侵袭数量、迁移距离。Western blot检测胰腺癌细胞中TLR4、NF-κB、NF-κBp65、髓样分化因子88（MyD88）蛋白表达水平。统计学方法采用t检验、单因素方差分析、SNK-q检验。结果　MEG3组PANC-1细胞克隆形成率、侵袭数量及迁移距离均低于空白组和空转组，PANC-1细胞凋亡率高于空白组和空转组，差异均有统计学意义（F=23.070、99.390、23.980、265.000，均P<0.05）。MEG3组和抑制剂组PANC-1细胞中TLR4、NF-κB、NF-κBp65、MyD88蛋白表达均低于空白组和空转组（均P<0.05）。结论　lncRNA-MEG3能通过抑制TLR4/NF-κB基因表达，调控PANC-1细胞的迁移和侵袭。

关键词:

胰腺癌, 转染, 侵袭, 克隆, 凋亡