siRNA干扰膀胱恶性肿瘤B7-H3基因表达对T24细胞增殖活力的影响

Transfection of B7-H3 siRNA decreases the proliferation of the malignant bladder cancer cell line T24

目的通过siRNA敲低膀胱T24细胞B7-H3基因表达,观察该基因的mRNA水平及蛋白分子表达,探讨B7-H3基因低表达状态下膀胱肿瘤细胞的增殖活力。方法利用小干扰RNA(siRNA)技术构建B7-H3基因低表达的T24细胞系,RT-PCR、Western blot技术验证基因敲低效果,MTS实验检验T24肿瘤细胞增殖能力变化。结果以重组慢病毒为载体,成功构建稳定转染B7-H3基因的膀胱T24细胞系(T24/B7-H3-siRNA)。与T24/control siRNA对照组和T24/non-siRNA比较,T24/B7-H3-siRNA细胞的B7-H3 mRNA表达水平明显下调(P<0.05),B7-H3蛋白表达量也显著降低,差异有统计学意义(P<0.05)。T24/B7-H3-siRNA细胞增殖活力也显著降低(P<0.05)。结论通过siRNA干扰B7-H3基因表达,膀胱T24细胞增殖活力显著降,提示B7-H3分子参与了膀胱肿瘤的恶性增殖行为,可作为肿瘤免疫治疗的潜在靶点。

Objective To measure the mRNA and protein of B7-H3,and the proliferation in the bladder cancer cell line T24 transfected with B7-H3 siRNA.Methods T24 cells were transfected with B7-H3 siRNA.Knockdown efficacy was validated by real-time reverse transcription polymerase chain reaction(RT-PCR)and Western blot.Cell proliferation was detected by MTS assay.Results Compared with T24 cells transfected with control siRNA or treated with blank control,T24 cells transfected with B7-H3 siRNA showed the significantly downregulated mRNA and protein levels of B7-H3,and significantly decreased proliferative rate(P<0.05).Conclusion Knockdown of B7-H3 in T24 cells by siRNA transfection significantly decreases cell proliferation,suggesting the involvement of B7-H3 in the malignant proliferation of bladder cancer cells.B7-H3 is believed as a potential target for bladder cancer immunotherapy.