TPX2对人肺癌细胞增殖、凋亡及cleavedcaspase-3表达的影响

目的 探讨Xklp2靶蛋白(TPX2)对人肺癌细胞增殖、凋亡及活化的半胱氨酸天冬氨酸蛋白酶3(cleaved caspase-3)表达影响。方法 培养人肺癌细胞A549,分为Control组(不进行细胞转染)、siRNA-NC组(细胞转染TPX2 siRNA control)、TPX2 siRNA组(细胞转染TPX2 siRNA),采用实时荧光定量PCR (qPCR)和Western blot检测TPX2 siRNA的转染效果。取转染后各组细胞,采用噻唑蓝检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Western blot检测cleaved caspase-3蛋白表达情况。结果 TPX2 siRNA组中,TPX2 mRNA、蛋白水平和细胞光密度(OD)值均明显低于Control组和siRNA-NC组,细胞凋亡率、细胞中cleaved caspase-3蛋白表达水平均明显高于Control组和siRNA-NC组,差异均有统计学意义(P值均<0.05);而siRNA-NC组中,TPX2 mRNA、蛋白水平及细胞OD值、细胞凋亡率、细胞中cleaved caspase-3与Control组比较,差异均无统计学意义(P值均＞0.05)。结论TPX2 siRNA能够干扰人肺癌细胞A549中TPX2的表达。下调TPX2的表达能够抑制人肺癌细胞A549增殖,促进caspase-3活化,促进人肺癌细胞A549的凋亡。

Effects of TPX2 on proliferation,apoptosis and activated cysteine containing aspartate proteinase-3 expression of human lung cancer cells

Objective To investigate targeting protein for Xklp2(TPX2) on human lung cancer cells proliferation, apoptosis and expression of activated cysteine containing aspartate proteinase-3 (cleaved caspase-3).Methods The human lung cancer cells A549 was cultured, divided into control group (no transfection), siRNA-NC group (cells transfected with siRNA control) and TPX2 siRNA group (cells transfected with TPX2 siRNA). Real-time fluorescence quantitative PCR (qPCR) and Western blot were used to detect the transfection effect of TPX2 siRNA. The cells of each group were taken after transfection, the cell proliferation was detected by thiazolium (MTT), cell apoptosis was detected by flow cytometry, and the expression of cleaved caspase-3 protein was detected by Western blot.Results In TPX2 siRNA group, the TPX2 mRNA and protein levels and the optical density values were significantly lower than those in control group and siRNA-NC group, the rate of cell apoptosis and expression of cleaved caspase-3 protein levels were significantly higher than those in control group and siRNA-NC group, the differences were statistically significant (all P values<0.05). In the siRNA-NC group, the TPX2 mRNA and protein levels, cell apoptosis rate, optical density value and cleaved caspase-3 had no significant differences compared with control group (P>0.05).Conclusions TPX2 siRNA can interfere with the expression of TPX2 in human lung cancer cells A549, interfering the expression of TPX2 can inhibit the proliferation of human lung cancer cells A549, it promotes the activation of caspase-3 and the apoptosis of human lung cancer cells A549.