NIRF基因通过PI3K/Akt信号通路诱导人肺癌细胞凋亡

NIRF gene induced apoptosis of human lung cancer cells through PI3K/Akt signaling pathway

目的探究NIRF基因对人肺癌细胞凋亡的影响以及机制研究。方法使用重组质粒siRNANIRF(siRNA-NIRF组)和空载体(siRNA-NC组)转染人肺癌细胞A549,以未转染细胞为对照(Control组),免疫印迹试验(Western blot)检测转染效果;噻唑蓝(MTT)检测转染细胞增殖情况,使用流式细胞术检测转染细胞凋亡状况,蛋白免疫印迹(Western blot)检测细胞中蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、磷脂酰肌醇-3激酶(PI3K)、磷酸化PI3K(p-PI3K)蛋白的表达量。结果转染重组质粒siRNA-NIRF后,细胞中NIRF蛋白的表达量显著低于对照组(P<0.05);与对照组相比,siRNA-NIRF组细胞的存活率显著降低(P<0.05),凋亡率显著增高(P<0.05),siRNA-NC组无明显差异(P>0.05);与对照组相比,siRNA-NIRF组和siRNA-NC细胞中Akt、PI3K蛋白的含量无明显变化(P>0.05),但siRNA-NIRF组细胞中p-Akt和p-PI3K的含量显著低于对照组(P<0.05)。结论干扰NIRF基因可抑制肺癌细胞A549增殖,促进其凋亡,其作用机制是影响PI3K/Akt信号通路的关键因子的表达量。

Objective To study the effect and mechanism of NIRF gene on the apoptosis of human lung cancer cells. Methods The lung cancer cells A549 was transfected with siRNA-NIRF( the siRNA-NIRF group)and empty vector( the siRNA-NC group). The untransfected cells were taken as the control group. The NIRF expression was detect by Western blot. Thiazolyl blue( MTT) was used to detect the proliferation of transfected cells,and the apoptosis was detect by flow cytometry. Western blot was used to detect the expression of protein kinase B( Akt),phosphorylated Akt,phosphatidylinositol-3 kinase( PI3 K) and phosphorylated PI3 K protein. Results The expression of NIRF protein in the siRNA-NIRF group was significantly lower than that in the control group( P < 0. 05).Compared with the control group,the survival rate of lung cancer cells in the siRNA-NIRF group was significantly lower( P < 0. 05) and the apoptosis rate was significantly increased( P < 0. 05). The siRNA-NC group showed no significant difference( P > 0. 05). Compared with the control group,the expression of Akt,PI3 K protein in the siRNA-NIRF group and siRNA-NC cells had no significant change( P > 0. 05),but the levels of p-Akt and p-PI3 K in the siRNA-NIRF group were significantly lower than those in the control group( P < 0. 05). Conclusion NIRF gene can inhibit proliferation and promote apoptosis of lung cancer cell A549. The mechanism of NIRF gene affects the expression of the key factor of PI3 K/Akt signaling pathway.