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目的研究siRNA干扰沉默膀胱癌肝素酶基因对T24细胞增殖及MMP-2表达的影响方法体外化学合成肝素酶特异性小干扰RNA(siRNA),用脂质体2000转染至T24膀胱移行细胞癌细胞内,常规分为正常对照组、空白对照组、阴性对照组和转染实验组进行实验,用噻唑蓝(MTT)法检测细胞增殖情况,反转录聚合酶链法(RT-PCR)检测肝素酶基因的表达,免疫细胞化学方法检测MMP-2的表达。结果RNA干扰沉默T24膀胱移行细胞癌细胞肝素酶基因后,T24细胞增殖活力下降,三个不同干扰组细胞活力分别下降了(28.55±3.66)%(、27.20±3.25)%和(37.29±4.82)%。肝素酶mRNA表达下降,膀胱移行细胞癌细胞中MMP-2(明胶酶A)的表达亦明显下降(P均<0.05)。结论RNA干扰肝素酶基因后,细胞活力及肝素酶mRNA表达水平下降并且明显抑制膀胱癌细胞中MMP-2的表达。
Objective To investigate the effect of silencing bladder cancer heparinase gene on proliferation and MMP-2 expression of T24 cells by siRNA interference.Methods Heparinase-specific small interfering RNA (siRNA) was synthesized in vitro and transfected into T24 bladder transitional cell The cells were divided into normal control group, blank control group, negative control group and transfected experimental group routinely. The proliferation of cells was detected by MTT assay, and the reverse transcriptase-polymerase chain reaction (RT-PCR) Heparinase gene expression was detected by immunocytochemistry to detect the expression of MMP-2. Results RNA interference silenced T24 bladder cancer cell heparanase gene, T24 cell proliferation activity decreased, three different interference group decreased cell viability (28.55 ± 3.66)% (27.20 ± 3.25)% and (37.29 ± 4.82 )%. Heparinase mRNA expression decreased, and the expression of MMP-2 (gelatinase A) in bladder transitional cell carcinoma cells was also significantly decreased (all P <0.05). Conclusion RNA interference of heparinase gene, cell viability and heparinase mRNA expression decreased and significantly inhibited the expression of MMP-2 in bladder cancer cells.