Cai Jundong, Huang Jiuning, Wang Wulong, Zeng Jing, Wang Ping
Department of Radiotherapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin 300000, People's Republic of China.
Department of Radiotherapy, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264000, Shandong, People's Republic of China.
Cancer Manag Res. 2020 Nov 13;12:11597-11609. doi: 10.2147/CMAR.S274192. eCollection 2020.
To investigate whether miR-124-3p regulates the fibroblast growth factor 2 (FGF2)-epidermal growth factor receptor (EGFR) pathway by targeting MGAT5 to affect the pemetrexed resistance in lung adenocarcinoma cells.
PC9-MTA and H1993-MTA anti-pemetrexed lung adenocarcinoma cell lines were constructed. The cell viability of anti-pemetrexed and parent lung adenocarcinoma cells was analyzed using MTS assay and reverse transcription PCR to determine the expression of miR-124-3p. CCK8 assay, colony formation assay, and flow cytometry were used to determine cells' proliferation and apoptosis. FGF2-EGFR signaling pathway-related proteins and MGAT5 protein expression were quantified by Western blotting. The target relationship between miR-124-3p and MGAT5 was verified by double luciferase assay. A nude mouse model with a transplanted tumor was established using the anti-pemetrexed lung adenocarcinoma cells. Tumor volume and weight were determined, and the apoptosis of tumor cells was observed.
The half-maximal inhibitory concentration of pemetrexed in anti-pemetrexed lung adenocarcinoma cells was higher than that in parent lung adenocarcinoma cells, and the expression of miR-124-3p in the anti-pemetrexed cells was lower than that of the parent cells. In the miR-124-3p overexpression group, MGAT5 silencing group, and miR-124-3p+MGAT5 overexpression group, compared with the control group, the proliferation ability of cells and tumors was markedly reduced; their apoptosis rates were increased significantly; expression levels of FGF2 and p-EGFR/EGFR were decreased; and the growth rate and tumor volume and mass were reduced; however, the opposite results were obtained in the miR-124-3p silencing group (p<0.05).
miR-124-3p may inhibit the FGF2-EGFR pathway by targeting MGAT5 to decrease pemetrexed resistance in lung adenocarcinoma cells.
探讨miR-124-3p是否通过靶向N-乙酰葡糖胺转移酶V（MGAT5）调控成纤维细胞生长因子2（FGF2）-表皮生长因子受体（EGFR）通路，从而影响肺腺癌细胞对培美曲塞的耐药性。
构建PC9-MTA和H1993-MTA耐培美曲塞肺腺癌细胞系。采用MTS法分析耐培美曲塞和亲本肺腺癌细胞的细胞活力，并用逆转录PCR法检测miR-124-3p的表达。采用CCK8法、集落形成试验和流式细胞术检测细胞增殖和凋亡情况。通过蛋白质印迹法对FGF2-EGFR信号通路相关蛋白和MGAT5蛋白表达进行定量分析。通过双荧光素酶报告基因检测验证miR-124-3p与MGAT5之间的靶向关系。用耐培美曲塞肺腺癌细胞建立移植瘤裸鼠模型。测定肿瘤体积和重量，并观察肿瘤细胞凋亡情况。
耐培美曲塞肺腺癌细胞中培美曲塞的半数抑制浓度高于亲本肺腺癌细胞，耐培美曲塞细胞中miR-124-3p的表达低于亲本细胞。在miR-124-3p过表达组、MGAT5沉默组和miR-124-3p+MGAT5过表达组中，与对照组相比，细胞和肿瘤的增殖能力明显降低；凋亡率显著升高；FGF2和p-EGFR/EGFR的表达水平降低；生长速率以及肿瘤体积和质量减小；然而，miR-124-3p沉默组则得到相反的结果（p<0.05）。
miR-124-3p可能通过靶向MGAT5抑制FGF2-EGFR通路，从而降低肺腺癌细胞对培美曲塞的耐药性。
