目的：探讨RNA干扰斑点型POZ蛋白（SPOP）基因表达对人膀胱癌T24细胞生物学行为的影响。方法采用阳离子脂质体转染试剂Lipofectamine 2000将靶向沉默SPOP基因的小干扰RNA（ siRNA）序列siRNA1和siRNA2分别转染至T24细胞株（ siRNA1组和siRNA2组），设转染随机序列的T24细胞为对照组，采用实时定量逆转录聚合酶链反应（ qRT⁃PCR）和Western blotting分别检测各组转染48 h后的SPOP mRNA及蛋白水平，细胞增殖活性检测试剂盒（ CCK⁃8）检测细胞体外增殖活力，Transwell迁移试验及划痕试验观察各组的迁移能力。结果对照组、siRNA1组和siRNA2组的SPOP mRNA相对表达量为1�012±0�025、0�281±0�023和0�274±0�053，蛋白相对表达量为0�712±0�153、0�358±0�073和0�317±0�084，siR⁃NA1组和siRNA2组的SPOP mRNA和蛋白水平均低于对照组（ P＜0�05），siRNA1组和siRNA2组SPOP mRNA和蛋白水平的差异无统计学差异（ P＞0�05）；与对照组比较， siRNA1组和 siRNA2组的增殖活性升高，差异有统计学意义（ P＜0�05），且siRNA1组和siRNA2组的增殖率随转染时间的延长而增加（ P＜0�05）；Transwell迁移试验结果显示siRNA1组和siRNA2组的穿膜细胞数分别为（128�6±9�3）个和（134�5±8�6）个，均高于对照组的（92�5±8�4）个，差异有统计学意义（P＜0�05）；划痕试验结果显示siRNA1组和siRNA2组的划痕愈合率为（89�1±4�5）％和（93�7±5�1）％，均高于对照组的（32�6±2�8）％，差异有统计学意义（ P＜0�05）。结论 SPOP与膀胱癌的恶性行为有关，可能发挥抑癌基因的作用，如抑制增殖及迁移。%Objective To investigate the effect of speckle⁃type POZ protein (SPOP) on the biological behavior of bladder cancer T24by small interfering RNA ( siRNA) . Methods The sequences of siRNA⁃1 and siRNA⁃1 together with LipofectamineTM 2000 were transfected into T24 cell lines to silence the expression of SPOP. T24 cells were divided into negative control group, siRNA1 and siRNA2 group. Quantitative real time polymerase chain reaction ( qRT⁃PCR) and Western blotting method were used to detect the mR⁃NA and protein levels of SPOP in each group at 48 h after transfection. Cell proliferation assay kit ( CCK⁃8) was used to check the cell proliferation activity and the proliferation and migration were observed by Transwell migration assay and wound healing assay. Results The relative expression of SPOP mRNA were 1�012±0�025, 0�281±0�023 and 0�274±0�053 in control group, siRNA1 group and siRNA2 group. The relative expression of SPOP protein were 0�712 ± 0�153, 0�358 ± 0�073 and 0�317 ± 0�084 in control group, siRNA1 group and siRNA2 group. Both the mRNA and protein levels of SPOP were lower in siRNA1 group and siRNA2 group than control group ( P<0�05) . There was no significant difference in mRNA SPOP and protein levels between siRNA1 and siRNA2 groups ( P>0�05) . Compared with the control group, the proliferation activity of siRNA1 group and siRNA2 group were significantly in⁃creased, and the proliferation rates of siRNA2 and siRNA1 groups increased with the prolongation of transfection time ( P<0�05) . Re⁃sults of Transwell migration test showed that the number of membrane penetrating cells were 128�6 ± 9�3 and 134�5 ± 8�6 in siRNA1 group and siRNA2 group, higher than 92�5±8�4 in control group with statistical significance (P<0�05). Results of the scratch test showed that the wound healing rates were (89�1±4�5)% and (93�7±5�1)% in siRNA1 group and siRNA2 group, higher than (32�6 ±2�8)% in control group with statistical significance (P<0�05). Conclusion SPOP may play a tumor suppressor gene in the malig⁃nant behavior of bladder cancer via inhibiting proliferation and migration.
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