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目的探讨核小体结合蛋白(NSBP1)对人膀胱癌细胞增殖的影响。方法采用脂质体转染法将NSBP1基因转入膀胱癌EJ细胞系,分别通过CCK8、流式细胞仪、Real-Time PCR及West-ern blot法观察EJ细胞的增殖,细胞周期时相的分布、凋亡以及CyclinD1、CyclinB1、Bcl-2蛋白的表达。结果 Real-Time PCR及Western blot结果显示经转染后EJ细胞的NSBP1显著地受到抑制(P<0.01)。CCK8结果提示EJ细胞转染后随着时间的延长,抑制率越大(P<0.01)。流式细胞仪检测发现,经转染干预后EJ细胞G2期比例增加(P<0.01),而凋亡细胞并没有显著变化(P>0.05)。West-ern blot结果再次验证周期蛋白CyclinB1显著下调(P<0.05),而CyclinD1没有统计学意义(P>0.05),凋亡蛋白Bcl-2也没有统计学意义(P>0.05)。结论 NSBP1是通过调控周期蛋白CyclinB1进而阻断或延缓细胞周期,而非通过细胞凋亡抑制膀胱癌细胞增殖。NSBP1在膀胱癌细胞增殖中起着重要作用。
Objective To investigate the effect of nucleosome binding protein (NSBP1) on the proliferation of human bladder cancer cells. Methods NSBP1 gene was transfected into bladder cancer EJ cell line by lipofection method. The proliferation and cell cycle distribution of EJ cells were observed by CCK8, flow cytometry, Real-Time PCR and West-ern blot respectively , Apoptosis and expression of CyclinD1, CyclinB1, Bcl-2 protein. Results The results of Real-Time PCR and Western blot showed that NSBP1 of EJ cells was significantly inhibited after transfection (P <0.01). The result of CCK8 showed that the inhibition rate of EJ cells increased with time after transfection (P <0.01). The results of flow cytometry showed that the proportion of G2 cells in EJ cells increased (P <0.01) and the apoptotic cells did not change significantly (P> 0.05). West-ern blot results again showed that CyclinB1 was significantly down-regulated (P <0.05), while CyclinD1 was not statistically significant (P> 0.05). The apoptotic protein Bcl-2 was also not statistically significant (P> 0.05). Conclusion NSBP1 inhibits the proliferation of bladder cancer cells through blocking or delaying the cell cycle by regulating the expression of cyclinB1. NSBP1 plays an important role in the proliferation of bladder cancer cells.