MiR-144-3p对口腔鳞癌Cal27细胞增殖、侵袭、凋亡和PTEN/PI3K/AKT信号通路的影响

Effects of miR-144-3p on proliferation,invasion,apoptosis of oral squamous cell carcinoma Cal27 cells and PTEN/PI3K/AKT signaling pathway

目的探讨微小RNA(miR)-144-3p对口腔鳞癌CAL27细胞增殖、侵袭和凋亡的影响及其机制。方法将体外培养的CAL27细胞分为miR-144-3p模拟物(mimics)组、mimics阴性对照(NC-mimics)组、miR-144-3p抑制剂(inhibitor)组、inhibitor阴性对照(NC-inhibitor)组、miR-144-3p inhibitor加si-con组,miR-144-3p inhibito加si-PTEN组;采用RT-PCR检测各组细胞中miR-144-3p的表达;MTT法检测细胞增殖;Transwell小室实验检测细胞侵袭;流式细胞术检测细胞凋亡;双荧光素酶报告基因实验检测miR-144-3p与PTEN的靶向关系;Western blot检测PTEN、PI3K以及p-AKT的表达。结果miR-144-3p过表达后,细胞增殖、侵袭能力、PI3K和p-AKT蛋白表达均明显升高,而细胞凋亡率、PTEN蛋白表达明显降低(P<0.05);PTEN是miR-144-3p的靶基因;与抑制miR-144-3p表达比较,同时抑制miR-144-3p和PTEN后细胞活力、侵袭细胞数显著升高(P<0.05),细胞凋亡率、PTEN蛋白水平显著降低(P<0.05),PI3K和p-AKT蛋白水平显著升高(P<0.05)。结论miR-144-3p具有促进口腔鳞癌CAL27细胞增殖、侵袭并抑制细胞凋亡的作用,其作用机制可能与靶向调控PTEN/PI3K/AKT信号通路有关。

Objective To investigate the effect of miR-144-3 p on proliferation,invasion and apoptosis of oral squamous cell carcinoma CAL27 cells and its mechanism.Methods The CAL27 cells cultured in vitro were divided into miR-144-3 p mimics group,mimics negative control group(NC-mimics)group,miR-144-3 p inhibitor group,and inhibitor negative control group(NC-inhibitor),miR-144-3 p inhibitor+si-con group,miR-144-3 p inhibitor+si-PTEN group.RT-PCR was used to detect the expression of miR-144-3 p in each group of cells.MTT method was used to detect cell proliferation.Transwell assays was used to detect cell invasion.Flow cytometry was used to detect apoptosis.Double luciferase reporter assay was used to detect the targeting relationship between miR-144-3 p and PTEN.Western blot was used to detect the expression of PTEN,PI3 Kand the p-AKT.Results After the overexpression of miR-144-3 p,cell proliferation,invasion ability,the expression of PI3 Kand AKT were significantly increased,while the apoptosis rate and PTEN protein expression were significantly reduced(P<0.05).PTEN is a target gene of miR-144-3 p.Compared with the inhibition of miR-144-3 p,the cell viability and the number of invasive cells were significantly increased(P<0.05),and the apoptosis rate and PTEN protein level were significantly reduced(P<0.05),PI3 Kand p-AKT protein levels were significantly increased(P<0.05)after inhibiting both miR-144-3 p and PTEN.Conclusion MiR-144-3 p can promote the proliferation,invasion and inhibit apoptosis of oral squamous cell carcinoma CAL27 cells,and its mechanism may be related to the targeted regulation of PTEN/PI3 K/AKT signaling pathway.