| 龚 程,高明悦,孟俊宏,彭 聪,徐 健,刘杜先.circ-CPA4调节miR-140-3p/SGK1轴对非小细胞肺癌细胞增殖、凋亡、迁移和侵袭的影响[J].,2024,(13):2455-2462 |
| circ-CPA4调节miR-140-3p/SGK1轴对非小细胞肺癌细胞增殖、凋亡、迁移和侵袭的影响 |
| Effects circ-CPA4 on the Proliferation, Apoptosis, Migration and Invasion of Non-Small Cell Lung Cancer Cells by Regulating the miR-140-3p/SGK1 Axis |
| 投稿时间：2024-01-26 修订日期：2024-02-22 |
| DOI：10.13241/j.cnki.pmb.2024.13.010 |
| 中文关键词: circ-CPA4 miR-140-3p SGK1 非小细胞肺癌 细胞增殖 细胞凋亡 迁移 侵袭 |
| 英文关键词: circ-CPA4 miR-140-3p SGK1 Non-small cell lung cancer Cell proliferation Cell apoptosis Migration Invade |
| 基金项目:江苏省卫生计生委医学科研项目（H20160181） |
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| 中文摘要: |
| 摘要 目的：探讨环状RNA羧肽酶A4（circ-CPA4）调节miR-140-3p/血清和糖皮质激素诱导的蛋白激酶-1（SGK1）轴对非小细胞肺癌（NSCLC）细胞增殖、凋亡、迁移和侵袭的影响。方法：将H1299细胞分为5组：Ct组、si-NC组、si-circ-CPA4组、si-circ-CPA4+inhibitor-NC组、si-circ-CPA4+miR-140-3p inhibitor组。采用实时荧光定量逆转录聚合酶链反应（qRT-PCR）检测circ-CPA4、miR-140-3p表达；流式细胞术检测细胞凋亡；细胞计数试剂盒（CCK-8）法检测细胞增殖；划痕愈合实验检测细胞迁移；Transwell检测细胞侵袭；免疫印迹检测SGK1、半胱氨酸蛋白酶3（Caspase-3）、细胞周期蛋白D1（CyclinD1）、基质金属蛋白酶2（MMP-2）蛋白表达；双荧光素酶报告基因实验检测circ-CPA4与miR-140-3p、miR-140-3p与SGK1的靶向关系。将上述5组细胞悬液分别皮下注射到裸鼠体内，30天后，处死裸鼠并分离肿瘤，称量肿瘤质量。结果：在NSCLC组织和细胞中circ-CPA4、SGK1蛋白高表达，miR-140-3p低表达。si-circ-CPA4组与Ct组、si-NC组比较，circ-CPA4、划痕愈合率、OD450值、侵袭细胞数、SGK1、CyclinD1、MMP-2蛋白表达及裸鼠体内肿瘤质量显著降低，miR-140-3p表达、细胞凋亡率、Caspase-3蛋白表达水平显著升高（P＜0.05）；与si-circ-CPA4+inhibitor-NC组比较，si-circ-CPA4+miR-140-3p inhibitor组对应指标变化趋势与上述相反（P＜0.05）；沉默circ-CPA4可靶向下调miR-140-3p表达，下调miR-140-3p可靶向促进SGK1蛋白表达。结论：沉默circ-CPA4通过上调miR-140-3p来抑制SGK1蛋白表达，从而抑制NSCLC细胞增殖、迁移、侵袭，并促进细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of circular RNA carboxypeptidase A4(circ-CPA4) on proliferation, apoptosis, migration and invasion of non-small cell lung cancer (NSCLC) cells by regulating miR-140-3p/serum and glucocorticoid-induced protein kinase-1 (SGK1) axis. Methods: H1299 cells were grouped into five groups: Ct group, si-NC group, si-circ-CPA4 group, si-circ-CPA4 combined with inhibitor-NC group and si-circ-CPA4 combined with miR-140-3p inhibitor group. The real time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the expressions of circ-CPA4 and miR-140-3p. The flow cytometry was performed to detect cell apoptosis. The cell counting Kit (CCK-8) assay was performed to detect cell proliferation. Migration was detected by scratch-healing experiment. Invasion was detected by Transwell. The western blot was performed to detect the expressions of SGK1, cysteine protease 3 (Caspase-3), cyclin D1 (CyclinD1), and matrix metalloproteinase 2 (MMP-2) proteins. The targeting relationship between circ-CPA4 and miR-140-3p, miR-140-3p and SGK1 were detected by dual-luciferase reporter gene assay. The cell suspensions of the above five groups were subcutaneously injected into nude mice respectively, and after 30 days, the nude mice were sacrificed, the tumors were separated, and the tumor mass was weighed. Results: In NSCLC tissues and cells, circ-CPA4 and SGK1 proteins were highly expressed, and miR-140-3p was lowly expressed. Compared with Ct group and si-NC group, the circ-CPA4, scratch healing rate, OD450 value, number of invasive cells, SGK1, CyclinD1, MMP-2 protein expressions and tumor mass in nude mice were significantly decreased in si-circ-CPA4 group, the expression of miR-140-3p, the rate of apoptosis, and the protein expression of Caspase-3 were increased significantly (P<0.05). Compared with the si-circ-CPA4 combined with inhibitor-NC group, the change trend of the corresponding indicators in the si-circ-CPA4 combined with miR-140-3p inhibitor group was opposite to the above (P<0.05). The silencing circ-CPA4 could target down-regulate miR-140 -3p expression, down-regulation of miR-140-3p can target and promote SGK1 protein expression. Conclusion: Silencing circ-CPA4 inhibits SGK1 protein expression by up-regulating miR-140-3p, thereby inhibiting proliferation, migration and invasion of NSCLC cells, and promoting cell apoptosis. |
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