Bao D, Yuan R-X, Zhang Y
Department of Ultrasonography, Shanxian Central Hospital, Heze, Shandong, China.
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6632-6638. doi: 10.26355/eurrev_202006_21649.
To explore the effects of long non-coding ribonucleic acid (lncRNA) maternally expressed gene 3 (MEG3) on proliferation and apoptosis of gallbladder cancer (GBC) cells and its molecular mechanism.
The relative expression level of lncRNA MEG3 in GBC cell lines was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The lncRNA MEG3 expression plasmids were constructed, the cell proliferation ability was detected via Cell Counting Kit-8 (CCK-8) assay and colony formation assay, and the apoptosis was detected via flow cytometry. The effects of lncRNA MEG3 expression on endoplasmic reticulum stress (ERS)-related proteins were determined using Western blotting, and the changes in nuclear factor-κB (NF-κB) protein in the nucleus were determined after overexpression of lncRNA MEG3.
The expression of lncRNA MEG3 in three kinds of GBC cell lines was lower than that in human immortalized normal biliary epithelial cells (p<0.05). The results of CCK-8 assay and colony formation assay showed that overexpression of lncRNA MEG3 significantly reduced the proliferation rate and colony formation ability of GBC-SD cells compared with negative control (NC) group (p<0.05, p<0.05). According to the results of flow cytometry, the apoptosis rate was higher in lncRNA MEG63 overexpression group compared with that in NC group (p<0.05). Moreover, the ERS-related proteins (MANF, GRP78, and caspase-3) were remarkably upregulated in lncRNA MEG63 overexpression group compared with those in NC group, indicating that ERS is activated by lncRNA MEG63 overexpression. The NF-κB signal in GBC cells was activated by lncRNA MEG3.
LncRNA MEG3 activates the NF-κB signal in GBC cells to affect the proliferation and apoptosis of GBC cells.
探讨长链非编码核糖核酸（lncRNA）母系表达基因3（MEG3）对胆囊癌细胞增殖和凋亡的影响及其分子机制。
通过定量逆转录-聚合酶链反应（qRT-PCR）检测lncRNA MEG3在胆囊癌细胞系中的相对表达水平。构建lncRNA MEG3表达质粒，采用细胞计数试剂盒-8（CCK-8）法和集落形成试验检测细胞增殖能力，通过流式细胞术检测细胞凋亡情况。采用蛋白质印迹法检测lncRNA MEG3表达对内质网应激（ERS）相关蛋白的影响，在lncRNA MEG3过表达后检测细胞核中核因子-κB（NF-κB）蛋白的变化。
lncRNA MEG3在三种胆囊癌细胞系中的表达低于人永生化正常胆管上皮细胞（p<0.05）。CCK-8法和集落形成试验结果显示，与阴性对照组（NC）相比，lncRNA MEG3过表达显著降低了GBC-SD细胞的增殖率和集落形成能力（p<0.05，p<0.05）。流式细胞术结果显示，lncRNA MEG63过表达组的凋亡率高于NC组（p<0.05）。此外，与NC组相比，lncRNA MEG63过表达组中ERS相关蛋白（MANF、GRP78和caspase-3）显著上调，表明lncRNA MEG63过表达激活了ERS。lncRNA MEG3激活了胆囊癌细胞中的NF-κB信号。
lncRNA MEG3激活胆囊癌细胞中的NF-κB信号，影响胆囊癌细胞的增殖和凋亡。
