LncRNA MEG3靶向miR-181a-5p调控宫颈癌细胞放射敏感性

目的 研究LncRNA MEG3对宫颈癌细胞放射敏感性的影响,并探讨其作用机制。方法 运用qRT-PCR法检测放射抗性和放射敏感性宫颈癌细胞中LncRNA MEG3的表达;将过表达对照组(转染pcDNA 3.1)、过表达LncRNA MEG3组(转染pcDNA 3.1-LncRNA MEG3)、抑制miR-NC组(转染anti-miR-NC)、抑制miR-181a-5p组(转染anti-miR-181a-5p)、过表达LncRNA MEG3+过表达miR-NC组(共转染pcDNA 3.1-LncRNA MEG3和anti-miR-NC)、过表达LncRNA MEG3+过表达miR-181a-5p组(共转染pcDNA 3.1-LncRNA MEG3和anti-miR-181a-5p),均用脂质体法转染至SiHa细胞;克隆形成实验检测细胞的存活分数;流式细胞术检测细胞的凋亡率;双荧光素酶报告基因检测实验检测细胞的荧光活性;Western blot检测细胞中PTEN、p-Akt、Akt的蛋白表达。结果 与放射敏感组相比,放射抗性宫颈癌组织中LncRNA MEG3的表达明显降低(P<0.05),其表达量与宫颈癌细胞的放射敏感性呈正相关;过表达LncRNA MEG3、抑制miR-181a-5p均可显著增强宫颈癌细胞SiHa放射敏感性,促进凋亡(P<0.05);野生型LncRNA MEG3细胞的荧光活性受miR-181a-5p的抑制。过表达miR-181a-5p逆转了LncRNA MEG3对宫颈癌细胞放射增敏和促凋亡作用及对PTEN/Akt信号通路的调控。结论 长链非编码RNA LncRNA MEG3可增强宫颈癌细胞放射敏感性,其机制可能与靶向miR-181a-5p调控PTEN/Akt 信号通路有关,可为提高宫颈癌的预后提供新方向。

LncRAN MEG3 regulates the radiosensitivity of cervical cancer cells by targeting miR-181a-5p

Objective To evaluate the effect of long-chain non-coding RNA MEG3(LncRNA MEG3) on the radiosensitivity of cervical cancer cells, and to explore its underlying mechanism. Methods The expression of LncRNA MEG3 in cervical cancer cells was detected by qRT-PCR. In the overexpression control group (transfected with pcDNA 3.1), LncRNA MEG3 overexpression group (transfected with pcDNA 3.1-LncRNA MEG3), miR-NC inhibition group (transfected with anti-miR-NC), miR-181a-5p inhibition group (transfected with anti-miR-181a-5p), LncRNA MEG3+miR-NC overexpression group (co-transfected with pcDNA3.1-LncRNA MEG3 and anti-miR-NC), LncRNA MEG3+miR-181a-5p overexpression group (co-transfected with pcDNA 3.1-LncRNA MEG3 and anti-miR-181a-5p), all plasmids were transfected into SiHa cells by liposome method. The cell survival fraction was assessed by colony formation assay. The cell apoptosis rate was evaluated by flow cytometry. The cell fluorescence activity was assessed by dual luciferase reporter assay. The expression levels of PTEN, p-Akt and Akt proteins were detected by Western blot. Results Compared with the radiosensitive group, the expression of LncRNA MEG3 was significantly down-regulated in radiation-resistant cervical cancer tissues (P<0.05), and its expression level was positively correlated with the sensitivity of cervical cancer cells. Overexpression of LncRNA MEG3 or inhibition of miR-181a-5p could significantly enhance the irradiation sensitivity and promote the apoptosis of cervical cancer cell line SiHa (both P<0.05). The fluorescence activity of wild-type LncRNA MEG3 cells was inhibited by miR-181a-5p. Overexpression of miR-181a-5p reversed the irradiation sensitization and pro-apoptosis effect of LncRNA MEG3 and the regulation of the PTEN/Akt signaling pathway on cervical cancer cell. Conclusion LncRNA MEG3 can enhance the sensitivity of cervical cancer cells to radiation exposure, probably by targeting the miR-181a-5p and regulating the PTEN/Akt signaling pathway, which will provide a new direction for improving clinical prognosis of cervical cancer patients.