hsa_circ_0072309对非小细胞肺癌细胞增殖、迁移、侵袭和凋亡的影响及机制研究

Effect and mechanism of hsa_circ_0072309 on proliferation,migration,invasion and apoptosis of non-small cell lung cancer cells

目的非小细胞肺癌(non-small cell lung cancer,NSCLC)是呼吸系统常见的恶性肿瘤,临床表现无特异性。环状RNA(circular RNA,circRNA)在肿瘤进展和细胞功能中起重要作用;然而,NSCLC和hsa_circ_0072309之间的关系仍不清楚。本研究旨在探讨hsa_circ_0072309的分子机制及其在NSCLC细胞增殖、侵袭、迁移和凋亡中的作用。方法在NSCLC细胞A549和细胞H1975中分别转染siRNA对照质粒(si-NC)和hsa_circ_0072309 siRNA干扰质粒(si-circ_0072309),构建hsa_circ_0072309干扰瞬转细胞株和对照细胞株。CCK8实验检测干扰后细胞增殖变化;划痕实验检测干扰后细胞迁移变化;Transwell实验检测干扰后细胞侵袭变化;流式细胞术检测干扰后细胞凋亡变化。采用Western Blot检测hsa_circ_0072309干扰后,细胞A549中AKT、p-AKT、PI3K和p-PI3K蛋白水平表达变化。结果与si-NC组相比,si-circ_0072309组A549细胞增殖速率无明显差异,迁移和侵袭细胞数量均显著降低(P<0.05),细胞早期凋亡显著减少(P<0.05);H1975细胞增殖速率、迁移和侵袭细胞数量均显著降低(P<0.05),细胞早期凋亡显著减少(P<0.05)。Western Blot结果显示,与si-NC组相比,si-circ_0072309组A549细胞中PI3K/AKT信号通路相关蛋白AKT和PI3K蛋白水平表达无明显变化,而p-AKT和p-PI3K蛋白水平表达显著下调(P<0.05)。结论沉默hsa_circ_0072309可能通过抑制肺NSCLC中PI3K/AKT信号通路中AKT和PI3K磷酸化,从而抑制NSCLC增殖、迁移、侵袭和凋亡。

Objective Non-small cell lung cancer(NSCLC)is a common malignant tumor in the respiratory system,with non-specific clinical manifestations.Circular RNA(circRNA)plays an important role in tumor progression and cellular function;However,the relationship between non-small cell lung cancer and hsa_circ_0072309 is still unclear.The aim of this study is to explore the molecular mechanism of hsa_circ_0072309 and its role in the proliferation,invasion,migration,and apoptosis of non-small cell lung cancer cells.Methods siRNA control plasmids(si-NC)and hsa_circ:0072309 siRNA interference plasmids(si-circ_0072309)were transfected into non-small cell lung cancer cells A549 and H1975,respectively,in order to construct hsa_circ_0072309 interference transient cell lines and control cell lines.CCK8 experiment was used to detect changes in cell proliferation after interference,scratch experiment to detect changes in cell migration after interference,Transwell experiment to detect changes in cell invasion after interference,flow cytometry to detect changes in cell apoptosis after interference,and Western blot to detect changes in protein expression levels of AKT,p-AKT,PI3K,and p-PI3K in A549 cells after interference with hsa_circ_0072309.Results Compared with the si-NC group,there was no significant difference in the proliferation rate of A549 cells in the si-circ_0072309 group,but the number of migrating and invasive cells was significantly decreased(P<0.05),and the early apoptosis of cells was significantly reduced(P<0.05).The proliferation rate of H1975 cells,the number of migrating and invading cells,and the early apoptosis of H1975 cells were significantly decreased(P<0.05).Western blotting results showed that compared with the si-NC group,there was no significant change in the expression of PI3K/AKT signaling pathway related proteins AKT and PI3K protein levels in A549 cells of the si-circ_0072309 group,while the expression of p-AKT and p-PI3K protein levels was significantly down-regulated.Conclusion Silenced hsa_circ_0072309 inhibits cell proliferation,migration,invasion,and apoptosis by inhibiting AKT and PI3K phosphorylation in the PI3K/AKT signaling pathway in NSCLC cells.