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目的探讨磷脂酰肌醇-4-磷酸酶Ⅱ型(INPP4B)对宫颈癌细胞生长增殖、细胞周期及凋亡的影响。方法Western blotting检测常见的宫颈癌细胞系中INPP4B的基础表达水平,对INPP4B阴性的Si Ha、C33a细胞转染表达质粒LHCX-INPP4B,高表达INPP4B的Ca Ski细胞转染INPP4B特异性的siRNA,Western blotting检测细胞中INPP4B表达的变化。CCK-8、克隆形成实验检测INPP4B对宫颈癌细胞生长增殖的影响;Brd U染色检测细胞周期,Annexin V-FITC/PI双染后流式细胞仪检测细胞凋亡。结果 CCK-8和克隆形成实验显示,过表达INPP4B使Si Ha和C33a细胞的活性和克隆形成能力明显下降(P均<0.05);Brd U实验表明,转染INPP4B后Si Ha细胞和C33a细胞进入S期的比例减少(P=0.04,P=0.01)。与之相应,干扰INPP4B表达后Ca Ski细胞的生长增殖能力增强,进入S期的细胞比例增加(P<0.01)。但INPP4B表达的变化对Si Ha、C33a和Ca Ski细胞的凋亡无明显影响。结论 INPP4B可抑制宫颈癌细胞的生长增殖和细胞周期进程。
Objective To investigate the effects of phosphatidylinositol 4-phosphatase Ⅱ (INPP4B) on the growth, cell cycle and apoptosis of cervical cancer cells. Methods The basic expression levels of INPP4B in cervical cancer cell lines were detected by Western blotting. INPP4B-specific siRNA and siRNA were transfected into Si-Ha and C33a cells transfected with expression vector LHCX-INPP4B, and INPP4B-overexpressing Ca Ski cells. blotting was used to detect the changes of INPP4B expression in cells. CCK-8 was used to detect the effect of INPP4B on the growth and proliferation of cervical cancer cells. The cell cycle was detected by BrdU staining and the apoptosis was detected by flow cytometry with Annexin V-FITC / PI double staining. Results CCK-8 and clone formation assay showed that over-expression of INPP4B significantly decreased the activity and clonality of Si Ha and C33a cells (all P <0.05). BrdU assay showed that Si Ha cells and C33a cells transfected with INPP4B S phase decreased (P = 0.04, P = 0.01). Correspondingly, the ability of Ca Ski cells to proliferate and proliferate after interference with INPP4B expression increased, and the proportion of cells entering S phase increased (P <0.01). However, the changes of INPP4B expression had no significant effect on the apoptosis of Si Ha, C33a and Ca Ski cells. Conclusion INPP4B can inhibit the growth and proliferation of cervical cancer cells and cell cycle progression.