下调LOX基因对肾癌细胞增殖、凋亡及细胞周期的影响及机制研究

目的 探讨LOX对肾癌细胞增殖、凋亡及细胞周期的影响及其作用机制。方法 将si-con、si-LOX质粒分别转染至786-0细胞中，记为si-con组、si-LOX组，转染采用脂质体法。蛋白质印迹（Western Blot）检测赖氨酰氧化酶(LOX)、周期素依赖性激酶4（CDK4）、Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤/白血病-2（Bcl-2）、磷酸化蛋白激酶B（p-Akt）、蛋白激酶B（Akt）表达水平；四甲基偶氮唑盐比色法(MTT)检测细胞活力；流式细胞术检测细胞凋亡和细胞周期。结果 与正常肾小管上皮细胞HK-2相比，肾癌细胞786-0、GRC-1、A498中LOX表达水平显著升高（P＜0.001）。敲减LOX肾癌细胞786-0的活力显著降低（t=8.440，P＜0.001），786-0细胞的凋亡率显著升高（t=25.628，P＜0.001），G0/G1期细胞所占比例增加（t=7.211，P＜0.001），S期细胞所占比例减少（t=13.190，P＜0.001），786-0细胞中CDK4、Bcl2、p-Akt表达水平显著降低（t=15.660、13.914、12.349，P＜0.001），786-0细胞中Bax表达水平显著升高（t=22.057，P＜0.001）。结论 敲减LOX能抑制肾癌细胞增殖，促进细胞凋亡，影响细胞周期，其机制可能与PI3K/Akt信号通路有关，可为肾癌的治疗提供新思路。

Effect of LOX gene down-regulation on proliferation,apoptosis and cell cycle of renal cell carcinoma and its mechanism

To investigate the effects of LOX on proliferation, apoptosis and cell cycle of renal cell carcinoma and its mechanism. Methods The si-con and si-LOX plasmids were transfected into 786-0 cells, respectively, and recorded as si-con group and si-LOX group, and transfected by liposome method. Western blot was used to detect expressions of lysyl oxidase(LOX), cyclin-dependent kinase 4(CDK4), Bcl-2 related X protein(Bax), B-cell lymphoma/leukemia-2(Bcl-2), Phosphorylated protein kinase B(p-Akt), protein kinase B(Akt); tetramethylazozolium salt colorimetric assay(MTT) was used to detect cell viability; flow cytometry to detect apoptosis and cell cycle. Results Compared with normal renal tubular epithelial cells HK-2, LOX expression levels were significantly increased in renal cancer cells 786-0, GRC-1, and A498(P<0.001). The cell viability was significantly decreased(t=8.440, P<0.001), the apoptosis rate was significantly increased(t=25.628, P<0.001), the proportion of G0/G1 phase cells increased(t=7.211, P<0.001), the proportion of S phase cells decreased(t=13.190, P<0.001), and the expressions of CDK4, Bcl2, and p-Akt significantly decreased(t=15.660, 13.914, 12.349, P<0.001), the expression of Bax significantly increased of knock down LOX in renal cancer cell line 786-0(t=22.057, P<0.001). Conclusion Knockdown of LOX can inhibit the proliferation of renal cancer cells, promote cell apoptosis, and affect cell cycle. The mechanism may be related to PI3 K/Akt signaling pathway, which may provide new ideas for the treatment of renal cell carcinoma.