微小RNA-92a靶向PTEN/Akt信号通路对鼻咽癌细胞增殖与凋亡的影响

Effect of miR-92a targeting PTEN/Akt signaling pathway on proliferation and apoptosis of nasopharyngeal carcinoma cells

目的探讨微小RNA-92a(miR-92a)通过靶向PTEN/Akt信号通路对鼻咽癌HONE1细胞增殖与凋亡的调控作用。方法采用Lipofectamine脂质体法向HONE1细胞转染miR-92a抑制剂(miR-92a组)和阴性对照(NC组),设不行转染的HONE1细胞为对照组。转染成功后用实时荧光定量PCR(QPCR)检测miR-92a在不同细胞组中的表达情况;MTT法检测各组细胞的增殖率;流式细胞技术检测各组细胞的凋亡率;Western blotting实验检测PTEN/Akt信号通路中PTEN和p-Akt蛋白的表达量。结果转染48 h miR-92a组与对照组和NC组相比,miR-92a的表达量明显降低(P<0.05),而对照组和NC组的差异无统计学意义(P>0.05)。miR-92a组转染24、48、72、96 h的细胞增殖率分别为(84.51±2.74)%、(77.21±3.55)%、(62.07±3.57)%和(49.25±4.15)%,其中72、96 h的细胞增殖率低于对照组和NC组(P<0.05);miR-92a组转染48 h的细胞凋亡率为(46.12±1.79)%,高于对照组的(6.99±0.72)%和NC组的(8.42±0.81)%,差异有统计学意义(P<0.05);miR-92a组转染48 h PTEN和pAkt蛋白的表达水平分别为0.61±0.12和0.37±0.09,对照组分别为0.41±0.11和0.73±0.14,NC组分别为0.39±0.08和0.68±0.07,与对照组和NC相比,miR-92a组细胞的PTEN表达升高,p-Akt表达降低,差异有统计学意义(P<0.05)。结论通过抑制miR-92a的表达来抑制鼻咽癌细胞增殖且促进其凋亡,可能是通过负调控PTEN/Akt信号通路来实现的。

Objective To investigate the regulation of microRNA-92a （miR-92a） on the proliferation and apoptosis of naso- pharyngeal carcinoma cell line HONE1 by targeting PTEN/Akt signaling pathway. Methods The miR-92a inhibitor （miR-92a group） and the negative control （NC group） were transfected into HONE1 cells by Lipofectamine liposome, and the HONE1 cells without transfection were used as control group. QPCR was used to detect the expression of miR-92a in different cell groups after transfecfion. The cell proliferation rate and apoptotic rate were detected by MTT method and flow cytometry, respectively. The expressions of PTEN and p-Akt in PTEN/Akt signaling pathway were measured by Western blotting. Results 48 h after transfection, the expression of miR- 92a in miR-92a group was significantly lower than that in control group and NC group （P〈0. 05）, while the difference between control group and NC group were not statistically significant（ P〉0. 05）. The proliferation rates of miR-92a group after transfection of 24, 48, 72 and 96 h were （84. 51±2. 74）%, （77.21±3.55）%, （62. 07±3.57）% and （49. 25±4. 15）%, the data of 72 h and 96 h were lower than control group and NC group （P〈 0. 05 ）. The apoptotie rate of miR-92a group at 48 h after transfeetion was （46. 12± 1.79） %, higher than （6. 99-±0. 72）% of control group and （8.42±0. 81 ）% of NC group, and the difference was statistically significant （P〈0. 05）. The expression levels of PTEN and p-Akt were 0. 61±0. 12 and 0. 37±0.09 in miR-92a group, 0. 41±0. 11 and 0. 73 ±0. 14 in control group, and 0. 39±0. 08 and 0. 68±0. 07 in NC group at 48 h after transfection. Compared with control group and NC group, elevated level of PTEN expression and decreased level of p-Akt expression were observed in miR-92a group （P〈0. 05）. Conclusion Inhibiting the expression of miR-92a can inhibit the proliferation of nasopharyngeal carcinoma ceils and promote its apoptosis, which may be achieved by negative regulation of PTEN/Akt signaling pathway.